IMMUNOLOGY AND SEROLOGY

Introduction (WEEK 2/ LEC)

1st SEM, 2021

Introduction to Immunology COWPOX

● The study of organisms’ body protection
from foreign invading / disease causing
microorganisms.
“recognize - dispose” : neutralize
● The study of the molecules, cells, organs, and
systems responsible for the recognition and
disposal of foreign - antigen (proteins)
(nonself) material
● A science that deals with the immune system SMALLPOX
and the cell mediated and humoral aspects
suspended in fluid (antibodies) of immunity
and immune responses
IMMUNITY
● The condition of being resistant to
infection.
● VARIOLATION - inhaled the powdered form
small pox
● VACCINATION

HISTORY OF IMMUNOLOGY
● He described how he deliberately inserted
matter taken from a sore on the hand of a
diarymaid, into the arm of the boy by means
of two superficial incision.
● Barely penetrating the cutis,
each about half an inch long.
After 2 months inoculated by small pox
(matter) from patient

HISTORY OF IMMUNOLOGY
● In 1798, Edward Jenner (an English Physician)
demonstrated that a scab from a cowpox
lesion can replace smallpox scabs for
variolation.
● This procedure was called: Vaccination
(from ‘Vacca’ for cow in Latin).
● Eventually led to the eradication of smallpox
from the world in 1970s

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021
HISTORY OF IMMUNOLOGY
● The technique of vaccination against
smallpox quickly spread through the
world.
● In 1980, the World Health Assembly
officially declared "the world and its
peoples" free from endemic smallpox.
HISTORY AND MILESTONES IN iMMUNOLOGY
(Scientists and their Discoveries)
● Voltaire (1773)
○ Reported on Chinese practice of
variolation (inhalation on obtain a
crust from an infected smallpox
patient)
● Edward Jenner (1798)
○ Smallpox vaccination (milder from of
virus - cox pox you from small pox)
● Haeckel (1862)
○ Phagocytosis (engulf)
● Pasteur (1880-1881) (Father of Immunology)
○ Live, attenuated chicken cholera
and anthrax vaccines;
○ (1885) live attenuated vaccine for
rabies
○ Attenuated - make the pathogen less
virulent
● Elie Metchnikoff (1883-1905)
○ Cellular theory of immunity through
phagocytosis (Novel Price Winner)
○ Used a starfish model
● Von Behring, Kitasata (1890)
○ Humoral theory of immunity
proposed
○ Humoral - protein/components in the
fluid
○ Diphtheria and tetanus vaxx
● Robert Koch (1891)
○ Demonstration of cutaneous
hypersensitivity
○ Exaggerated immune response
● Gruber and Durham (1896)
○ Discover agglutination reactions
○ Formation of large complexes
● Ferdinand Widal (1896)
○ Devised agglutination reaction for
the diagnosis of typhoid fever
○ typhoid fever (S. typhii)
○ Widal test - for typhoid fever
● Paul Ehrlich (1900)
○ Antibody formation theory
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● Karl Landsteiner (1900)
○ Discovery of the ABO Blood Group
(blood types)
● P.Portier and Charles Richet (1902)
○ Immediate hypersensitivity
anaphylaxis
● Almorth Wright, Stephen Douglas, Joseph
Denys (1903)
○ Discovered opsonins and describe its
relation to phagocytosis
○ opsonins - proteins circulating in the
humarol system w/c coat the antigen
to be more easily recognize for
engulfment/phagocytosis
● Von Pirquet and Schick (1906)
○ Describe the relationship of immunity
and hypersensitivity
● Albert Calmette and Camille Guerin (1921)
○ 1st successful vaccine against
tuberculosis
● Pfeiffer (1894- 1895), Buchner
○ Complement (proteins) mediated
cytolysis
○ Complement - proteins that mediate
inflammation complete the action of
antibody by cytolysis enhance
antibody function killed & attenuated
form
● Jules Bordet
○ Received Nobel prize for his
pioneering works on complement
● Salk and Sabin
○ Development of polio vaccine
● George Snell (1903)
○ Worked out the genetics of the
murine major histocompatibility
complex and generated the congenic
strains needed for its biologic analysis
● Niel Jerne (1911)
○ Developed the hemolytic plaque
assay and several important
immunological theories including an
early version of clonal selection
● Peter Medawar (1915- 1987)
○ Made studies on acquire immunologic
tolerance
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

● Jean Dausset (1906) ● Luc Montaigner (1980)

○ An early pioneer in the study of MHC
or HLA
○ HLA - to conduct organ
transplantation should be match
● Baruj Benacerraf (1920)
○ Discovered immune response genes
and collaborated in the first
demonstartion of MHC restriction
● Rodney Porter (1920- 1985)
○ Worked out the polypeptide structure
of the antibody molecule, laying out
the groundwork of its analysis by
protein sequencing
● Rosalyn Yalow (1921)
○ Developed RIA (Radioimmunoassay)
of peptide hormones
● Alexander Flemming (1922)
○ Described the action of lysozymes
(organelles)
● James Gowan (1924)
○ Discovered that adaptive immunity is
mediated by lymphocytes
● Cesar Milstein (1927) and George Kohler
(1946)
○ Developed the technique of
monoclonal antibody formation
● Gerald Edelman (1929)
○ Made crucial discoveries about the
structure of Ig, including the
○ first complete sequence of Ab
molecule
● Michael Heidelberger (1888- 1991)
○ Developed the quantitative precipitin
assay.
○ In 1930- 1935, collaborated with
Kndall in conducting qualitative
precipitin assay studies on
antigen-antibody reactions
● Max Theiler (late 1930), Reed (1951)
○ Developed a vaccine against yellow
fever
● Susumu Tonegawa (1939)
○ Discovered the somatic recombination
of immunological receptor
○ genes that underlies the generation of
diversity in human and murine
antibodies and T cell receptors
○ Isolated retrovirus from a
non-immune deficient homosexual
man with lymphadenopathy and
called the virus lymphadenopathy
associated virus (LAV)
● Robert Gallo (1980)
○ Renamed the retrovirus as Human
Immunodeficiency Virus (causes AIDS)
● Mosmann (1986)
○ Th1 versus Th2 model of T-helper
function
● Edward Dounal Thomas and Joseph Murray
(1991)
○ Introduced the concepts of
transplantation
● 1996- 1998
○ Identification of toll-like receptors
○ toll-like receptors - pattern
recognition receptors involved in
direct phagocytosis
● Peter Doherty and Rolf Zinkernagel (1996)
○ Work on the mechanism of cellular
response mediated by T-cells towards
virally infected cells
● 2001
○ Discovery of FOX3p, the gene
directing regulatory T-cell
development
● Frazer
○ Discovery of the HPV vaccine

● Alic Isaacs and Jean Lindemann (1957)

○ Discovered interferon
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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

ADAPTIVE IMMUNITY
WEEK 2 PART 2-OVERVIEW AND TYPES OF
IMMUNITY
● A type of resistance that is characterized by
specificity for each individual pathogen, or
The Immune System microbial agent, and the ability to remember
(Overview and Types of Immunity) a prior exposure (w/memory & specificity)
● cellular/humoral mediated)
Innate Versus Adaptive Immunity
● Role of the Immune System
○ Defending the body against infections
○ Recognizing and responding to foreign
antigens
○ Defending the body against the
development of tumors
○ CELLS OF THE IMMUNE SYSTEM:
○ A deficiency or dysfunction of the
immune system can cause many
disorders
BODY DEFENSES: NATURAL IMMUNITY
RESISTANCE TO MICROBIAL DISEASE (Cells of the Innate Immune System)

TYPES OF IMMUNITY NATURAL IMMUNITY:

First Line of Defense
A .NATURAL IMMUNITY
● Ability of an individual to resist infections by
1. EXTERNAL DEFENSE MECHANISMS
means of normally present body functions
● Composed of structural barriers that prevent
● present since birth “mediated by WBC) most infectious agents from entering the body
● Innate, nonspecific, non-adaptive
B. ACQUIRED IMMUNITY A. Structural/ physical Barriers
● A reaction resulting from the invasion of a a.Intact skin
foreign substance. (mediated by lymphocytes. b.Mucous membranes of the respiratory and GI Tract
T&B cells) c.Ciliated Epithelium
● Adaptive & specific d.Lacrimal apparatus
NATURAL IMMUNITY e.Sweat, sebaceous glands
● No prior exposure is required
B. Mechanical Barriers
● The response lacks memory and specificity
a. Peristaltic movement of intestines
● Considered non-adaptive or nonspecific and
b. Shedding of cells
are the same for all pathogens or foreign
c. Coughing and sneezing
substances to which one is exposed
d. Flushing action of urine
● (cellular/humoral mediated response)

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

interacting with microbial membrane to form

C. Chemical Barriers
a. Acid pH- lactic acids and fatty acids, HCl,
Lactobacillus acidophilus in intestine and vagina
b. Lysozyme- attacks cell wall of microorganisms and
rendered it osmotically sensitive (found in tears
and saliva “IgA”)
c. Lactoferrin- in human milk
channels through which metabolites escapes
C. Interferons
○ group of molecules that limit the
spread of viral infections by blocking
translation of viral proteins
Types:
● Alpha IF - produced by leukocytes
● Beta IFN - produced fibroblasts
● Gamma IFN - produced by T.cytotoxic
T-cells

D. Complement
● Principal soluble mediator of inflammatory
response
● The major functions of complement are
opsonization, Chemotaxis, and lysis of cells.

E. Acute phase proteins

● serum molecules in which concentrations
increase rapidly at the onset of inflammatory
disease
a. CRP (C-reactive protein)
NATURAL IMMUNITY: b. Serum amyloid A
c. Mannose Binding Protein
Second line of Defense d. Alpha-1-antitrypsin
e. Haptoglobin
f. Ceruloplasmin
2.INTERNAL DEFENSE MECHANISMS
A. Humoral (found in fluid) g. Alpha-1-acid glycoprotein
● Complement h. Endogenous pyrogens
● Lysozyme
● Interferon INTERNAL DEFENSE MECHANISMS: Cellular
● Acute phase reactants
B. Cellular A. Neutrophils
● Mast cells ● 50-70% of circulating WBC’s
● Neutrophils ● Primary granules contain myeloperoxidase,
● Macrophages ACP, neutral proteinases, lysozymes, acid
hydrolases, B-glucuronidase and elastase
INTERNAL DEFENSE MECHANISMS: Humoral ● Secondary granules contain collagenase,
lysozyme, lactoferrin, plamsinogen activators
A.Physiologic factors and ALP
● Body temperature ● Capable of the process diapedesis
● Oxygen tension B. Eosinophils
● Hormonal balance ● 1-3% of circulating WBC’s
B. Basic polypeptides ● Increase in allergic reactions and parasitic
● Spermin - pH dependent polyamine found in diseases
semen; inhibits the growth of gram positive ● Reddish-orange granules contain ACP,
● Defensin - present in human neutrophil, are B-glucuronidase, aryl-sulfatase,
cationic proteins that kill microbes by phospholipase, peroxidase, histaminase,
aminopeptidase and ribonuclease
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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

C. Basophils
● less than 1% of circulating WBC’s
● Bluish-purple granules contains histamine,
eosipnophil chemotactic factor of anaphylaxis
and heparin
● Involved in immediate hypersensitivity
reactions
D. Mast cells
● can be found on connective tissues
● Granules contain ACP, ALP and protease
E. Monocytes
● largest WBC, constitutes 4-10% of circulating
WBC’s
● Possess grayish-blue cytoplasm and is ground
glass in appearance
Two modes of intracellular killing
● Granules contain peroxidase, ACP,
● Oxygen-dependent killing
arylsulfatase
○ by products of respiratory burst and
● Other type of granule contains
by halogenation of bacterial proteins
B-glucuronidase, lysozyme and lipase
catalysed by myeloperoxidase
F. Macrophages ● Oxygen-independent killing
● Larger version of monocytes on tissues ○ by lysosomal antibacterial substance
● There is increase in the number of ER, without the requirement of
lysosomes and mitochondria respiratory burst.
● Granules contain no peroxidase at all Pathways of killing Pathogens by Phagocytosis
compared with monocytes
● Monocyte-macrophage system functions in 1. Oxygen Dependent
microbial killing, tumoricidal activity, killing of
intracellular parasites, phagocytosis, ● Respiratory Burst-occurs when the
secretion of cell mediators and antigen cytoplasmic pseudopods enclosed the
presentation particle within a vacuole.
G. Dendritic cells
● Function is to phagocytosed antigen and
present it to T-helper cells
● Most potent phagocytic cell in the tissues

H. Natural killer cells

PHAGOCYTOSIS
● Engulfment of cells and particulate matter
by leukocyte, macrophage and other cells.

1. Physical contact between the WBC and the

foreign particle
2. Formation of phagosome 2. Oxygen independent
3. Fusion with lysosome
● Production of nitric oxide synthetase
4. Ingestion and release of debris to the
from oxidation of L-arginine by NO
outside
synthetase which is produced by
IFN-gamma activated cells

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

Inflammation
Effector Molecule Function
The overall reaction of the body to injury or invasion
by an infectious agent.
Cationic proteins Damage to microbial Major events:
(including cathepsin) membranes
1. Tissue damage cause release of vasoactive
and chemotactic factors that trigger a local
Lysoszyme Splits mucopeptide in increase in blood flow and capillary
bacterial call permeability.
wall
2. Permeable capillaries allow the influx of fluids
and cells
Lactoferrin Deprives proliferating 3. Phagocytes migrate to the site of the
bacteria of iron inflammation
4. Phagocytes and anti-bacterial exudates
destroy pathogen.
Proteolytic and Digestion of killed micro
hydrolytic enzymes oragnism

TYPES OF PHAGOCYTOSIS

INDIRECT DIRECT

Via opsonin receptors Via Pattern recognition

that recognize opsonins receptors that recognize
such as IgG, CRP and lipid and carbohydrate
C3b bound to sequences on
microorganism microorganisms

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

Cardinal signs

1. Rubor TWO ARMS OF ADAPTIVE IMMUNITY

2. Calor
3. Tumor
HUMORAL CELL MEDIATED
4. Dolor
5. Functio laesa 1. Mechanism Antibody Cell mediated
mediated
Major events associated with the process of
inflammation are: 2. Cell type B-lymphocytes T-lymphocytes
1. Increased blood supply to the infected area;
3. Mode of Antibodies in Direct cell to cell
2. Increased capillary permeability caused by action biologic fluid contact or
retraction of endothelial cells lining the soluble products
vessels; secreted by cells
3. Migration of white blood cells, mainly
neutrophils, from the capillaries to the 4. Function Primary defense Defense against
surrounding tissue; against viral and fungal
bacterial infections,
4. Migration of macrophages to the injured area
infections intracellular
organism, tumor
antigens and
ACQUIRED IMMUNITY graft rejection,
parasite
ACTIVE PASSIVE
5. Nature of Circulating Intracellular
1. Natural Active 1. Natural Passive infecting antigen extracellular organism
antigen
2. Artificial Active 2. Artificial Passive
6. Type of Acute pyogenic Chronic,
infection infection granulomatous
infection,
ACQUIRED IMMUNITY neoplasm,
fungal, parasitic
disease

7. Variant Ab-mediated Contact

hypersensitivities sensitivity,
, Autoimmunity allograft
rejection,
elimination of
tumors,
formation of
Chronic
granulomas

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021
The Immune System
▪ Lymphatic system
▪ Computerized war machine
▪ “defense department” of the body
At the end of this unit, the student will be able to:
• Classify and discuss the cells of the immune system
correctly.
• Outline and discuss the phylogeny and ontogeny of
the lymphoid system accurately.
• Differentiate B-lymphocytes from Tlymphocytes in
terms of its role in the adaptive immune response.
• Accurately explain the mechanisms of NK cell
cytotoxicity
• Enumerate and describe the different laboratory
identification of lymphocytes correctly
Cells of the Immune System
1. Myeloid lineage (mediated by the cells in the innate
immunity)
▫ The myeloid lineage eventually differentiates further
and matures to become platelets, erythrocytes,
monocytes or granulocytes.
2.Lymphoid lineage (mediates the adaptive
immunity)
▫ The lymphoid line will develop into two different
types, T and B cells, depending upon where they
complete their maturation, thymus or bone marrow
Major Functions of the Immune System
1. Block harmful agents
2. Seek-out invasive pathogens (recognizes)
3. Isolate and neutralize activity of that antigen
(dispose)
Acquired Immunity
● Type of resistance that is characterized by
specificity for each individual pathogen and
the ability to remember a prior exposure,
which results in an increased response upon
repeated exposure.
● Activates when exposed in foreign antigen
Part 1: Cells of the Adaptive Immune System
T cells, B cells, Natural killer cells
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Lymphocytes
▪ The key cell involved in adaptive immunity
▪ Represents approximately 20% of the circulating
WBC’s
▪ Has a large rounded nucleus which contains a dense
nuclear chromatin.
2 Major Stages of Lymphocyte Development
1. Antigen Independent Stage of Lymphopoiesis
● happens in bone marrow
● not stimulated by antigen/antigenic exposure
2. Antigen Dependent Stage of Lymphopoiesis
● create lymphocyte because of antigenic
exposure; stimulated by antigen
● happens in Secondary Lymphoid Organs
(spleen,lymph nodes etc. )
Cells of the Adaptive Immune System
▪ Lymphocytes
● T cells (61-81 %), B cells (10-20%), and natural
killer (NK) cells (10-15%)
● based on specific functions and the proteins
on their cell surfaces
▪ Clusters of differentiation (CD) aka Surface Markers
● proteins, or antigens, on cell surfaces can be
used to identify each lymphocyte
subpopulation
● As each antigen, or CD, was found, it was
assigned a number. (to be easily recognized)
● helps in identifying different lymphocytes
Surface Markers/CD Markers
▪ B cells (humoral mediated immune response;
mature and differentiated in the bone marrow))
● Role in antibody production
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021
● programmed to produce a unique antibody
molecule.
● It can be recognized by the presence of
membrane-bound antibodies of two types,
namely immunoglobulin M (IgM) and
immunoglobulin (IgD).
● Other surface proteins: CD19, CD21, and class
II major histocompatibility complex (MHC)
molecules
ADDED NOTES:
● B cells→ active→ form “blasts”--> become
plasma cells which will produce antibodies
▪ T cells (cell-mediated immune response;
differentiated in the Thymus)
● Produce cytokines (directly attacks cells; cell
to cell cytolytic activity)
● Stimulates B cells to produce antibodies,
● Assist in killing tumor cells or infected target
cells
● Help in regulation of both the innate and
adaptive immune response.
● Cell-mediated immunity.
● T cells can be distinguished according to their
unique functions:
○ Helper, cytolytic, and regulatory T
cells.
○ The subtypes can be identified by the
presence of the CD3 marker on their
cell surface, and either CD4, or CD8.
● T cells with CD4 receptor are mainly either
helper or regulatory cells
● CD8-positive (CD8+) population consists of
cytotoxic T cells.
● The ratio of CD4+ to CD8+ cells is
approximately 2:1 in peripheral blood
▪ Natural Killer cells
▫ A small percentage of lymphocytes do not
express the markers of either T cells or B cells.
▫ Have the ability to kill target cells without
prior exposure to them. (cell to cell contact)
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10
▫ Do not require the thymus for development
but appear to mature in the bone marrow
itself
The Lymphoid System
1. Primary/ Central Lymphoid Organs (Developmental
stage)
a. Bone (1 of the largest tissues in the body and fills
the core of all long flat bones)
▫ Main Marrow source of hematopoietic stem
cells
▫Center of antigen-independent
lymphopoiesis
b. Thymus
▫ small, flat bilobed organ found in thorax or
chest activity right below the thyroid gland
and overlying the heart
▫ An endocrine gland (where T cells mature)
▫ Cortex: where thymocytes(immature T cells;
precursor T cells) can be found (85% of
population of T-cells)
▫ Medulla: where mature T cells can be found
▫ Thymic stromal cells: include epithelial cells,
macrophages and dendritic cells
ADDED NOTES:
● B cells: 1st discovered in Birds (Bursa of
Fabricius) → equivalent organ is Bone marrow
2. Secondary/ Peripheral Lymphoid Organs
(Activation sites)
A. Encapsulated organs
i. Spleen
●Largest secondary lymphoid organ
that is found on the ULQ of the
abdomen just below the diagram
●Characterized as a large
discriminating filter, as it removes old
and damaged cells and foreign
antigens from the blood
ADDED NOTE:
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021
● Once matured they will migrate into the
Secondary Lymphoid Organ and be part of the
recirculating pool
Spleen
2 Main Types of Splenic Tissue:
⮚ Red Pulp: involved in Culling process (killing of
old/damaged WBC)
⮚ White Pulp : Contains lymphoid tissue that is
arranged around arterioles as PALS (Periarteriolar
Lymphoid Sheath)
• T cells are found near the central arteriole
• Naive B cells are found on primary follicles
• Activated B cells are found on Secondary Follicles in
the germinal centers
• Marginal zone contain macrophages/dendritic cells
that traps the antigen
ADDED NOTES:
● Macrophage (Ag presenting cells) → T cells
(activated) → secrete CYTOKINES that will
stimulate B cells to become plasma cells →
produce ANTIBODY
ii. Lymph Nodes
● Junctional filter of the lymphoid system
● Serve as central collecting points for lymph
fluid from adjacent tissues
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11
● Sizes may range from 1mm to 25 mm
● Cortex: B-cell area;
○ Contains primary follicles (Naïve B
cells), Specialized cells called follicular
dendritic cells.
○ secondary follicles that contains
germinal center (“BLAST”
transformation of B cell to plasma
cell)
○ Plasma cells, which actively secrete
antibody, and memory cells.
○ Generation of B-cell memory is a
primary function of lymph nodes
● Germinal center/Secondary Follicles (plasma
cells and activated B cells)
● Primary Follicles (Naive B cells)
Lymph Nodes
● Paracortex: T-cell area
○ the region between the follicles and the
medulla.
○ T lymphocytes (can only recognize foreign
antigens if it is bound to a phagocyte thru
MHC Classes) are in close proximity to antigen
presenting cells called interdigitating
cells/antigen presenting cells.
● Medulla: contains differentiated cells and APC’s
○ The medulla is less densely populated but
contains some T cells (in addition to B cells),
macrophages, and numerous plasma cells
Lymphocyte Traffic/Recirculation
▫ Movement of lymphocytes from blood to
lymphoid organs and back to the blood.
Site of Entry and Exit of Lymphocytes in the Lymph
node:
1. Afferent arteriole (towards lymph node, away from
blood)
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

2. Efferent arteriole (away from lymph nodes, towards

blood)

How will T and B cells find its way to the lymph

node?

■Through Post Capillary Traffic endothelial venule/

High-walled endothelial venule (HEV)

●Has adderesins and other adhesion

molecules: CD 32/ CD 102 marker

●T and B cells have homing receptors:

L-selectin of LAM (Leukocyte Adhesion
Molecule)

ADDED NOTES:

● Filtration of fluid from blood vessel is

important because it allows contact between
lymphocytes and the antigen
● Antigen Presenting Cells (macrophage, MHC)-
presents foreign materials to T cells

iii. Other Secondary Lymphoid tissues and Organs

▪ Mucosal Associated Lymphoid Tissue (MALT)

▫ Small masses of lymphoid tissue found in intestinal,
genitourinary tract and respiratory tract.
▪ Bronchus Associated Lymphoid Tissue (BALT)
▪ Gut associated lymphoid tissue (GALT)
▪ Cutaneous Associated Lymphoid Tissue (CALT)
▪ Tonsils (waldeyers’ ring)
▪ Peyer’s patches
▪ Bronchus associated lymphoid tissue
▪ Mammary glands
▪ Salivary glands
▪ Cutaneous immune system

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021
2. Pre-B Cells
Part II:The Adaptive Immune System
• T cell & B cell differentiation
• The Role lymphocytes in the Adaptive Immune
response
ADDED NOTES:
Stages of B Cell Differentiation
• Development of mature immunocompetent B cells
• Activation of B cells by antigen
• Differentiation of activated B cells into plasma cells,
which produce antibodies
B-cell Development: Antigen-Independent phase
1. Pro-B Cells
▫ Growth factors: E2A, EBF (early B-cell factor),
interferon regulatory factor (IFR8), and paired
box protein 5 (PAX5).
▫ Markers include CD 19, CD45R, CD43, TdT,
RAG-1 and RAG-2 enzyme
▫ The first step in the pro-B phase is the
rearrangement of genes that code for the
heavy and light chains of an antibody
molecule
ADDED NOTES:
3. Immature B Cells
● CD19- coreceptor that help regulate the B cell
differentiation and activation; PRESENT IN ALL
B CELL POPULATION/STAGES
● CD45- A glycoprotein that is PRESENT IN ALL
SURFACES OF HSCs
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13
● CD45R- found in the surface of Pro B cells that
are the LARGEST form
● TdT- (Terminal deoxyribonucleotide
Transferase) intracellular protein that is found
in this stage
● RAG 1 and RAG 2-( Recombination Activating
Genes) enzymes/genes involved in the gene
rearrangement.
● CD19, CD24,CD45 remain on the surface of
the B cells throughout the subsequent
developmental stage.
▫ Contains pre-B cell receptor that is made up
of 2 heavy chains and a surrogate light chain
whose function is to transmit signals to
prevent rearrangement of any other heavy
chain genes.
● Differentiation of B cell to Pro B cell into Pre B
cell upon successful rearrangement of your
heavy chains genes then these genes are
transcribed to make protein that will be part
of the antibody molecule
● The 1st heavy chains synthesized are the MU
chains for IgM which are accompanied by
unusual light chains known as surrogate light
chains (consist of short polypeptide chains
that are non covalently associated with each
other.
● If gene rearrangement does NOT work, the
Pre B cell development will be halted/
prevented thru Apoptosis.
● All Pre B cells that have MU chains
accompanied by surrogate light chains will
proceed to differentiation to subsequent B cell
stages.
● loss of CD43 and TdT
▫ Recognized by the appearance of complete
IgM molecules on their surface
▫ Specificity of the surface immunoglobulin to
be synthesized can already be predicted or
noted.
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

▫ Preexisting diversity of receptors ▫ Follicular B cells migrate to lymph

▫ Surface markers that can be seen include
receptors for complement components such
as C3d, CD21, CD40 and MHC class II
▫ B cells capable of producing antibody to
self-antigens are deleted in the marrow
through Apoptosis - Central tolerance
ADDED NOTES:
● Apoptosis happens in this stage
● MU chains is a component of IgM
● “Antibody will only bind into the Ag but the
complement will complete the activity of the
Antibody which is lysis of the Ag.”
● CD40 and MHC Class 2- are important for
interaction of B cells and T cells; MHC Class 2-
this are the ones that present antigens to the
T cells
● CD21- receptor for the breakdown product of
complement component which is C3,
important in the lysis of Antigen; enhances
the likelihood of contact between your B cells
and antigens
● Immature B cells that survive the selection
process will leave the Bone marrow and
proceed to the spleen and other Secondary
Lymphoid Organs.
● The B cell should recognize a self antigen;
Apoptosis of a self reacting B cell is very
important in the elimination of self reactive
receptors which is known as Central tolerance
4. Mature B cells
▪ In the spleen, immature B cells develop into
mature cells known as either marginal zone B
cells or follicular B cells
▫ Marginal B cells: remain in the
spleen (not exposed: Naive B cells)
- Respond quickly to any blood-borne
pathogens in contact
nodes and other secondary organs.
▪ Exhibit IgD and more IgM on their surface as
well as MHC Class II products
▪ If, a B cell is stimulated by antigen:
▫ Undergoes transformation to a blast
stage that eventually forms memory
cells and antibody-secreting plasma
cells
▫ This is known as the
antigen-dependent phase of B-cell
development.
-”humoral mediated immune
response”
ADDED NOTES:
Progenitor stem cell of Lymphoid lineage→
rearrangement of genes (Pro B cells) → successful
rearrangement; coded genes of 2 heavy chains
accompanied by light chains (Pre B cells) → complete
appearance of IgM molecule, where Apoptosis occurs
(Immature B cell) → presence of IgD; those survived
will further differentiate and release into the bone
marrow and go to the Secondary Lymphoid Organs
until antigen dependent stage occurs (Mature B cells)-
→ will continue to develop into Marginal or Follicular
B cells which recirculates in the body and constantly
survey for antigens for them to be activated →
“BLASTS” → will form plasma cells and memory cells
which secretes Antibodies
B-cell Development: Antigen-Dependent phase
4. Activated B cells

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
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▫ Exhibit CD25 on surface (exposed to antigen)
▫ CD25 in turn acts as receptor for IL-2
CD25- is found on BOTH Activated B cells and
Activated T cell and acts as receptor of IL-2
which are produced by T cells
5. Plasma
▫ Large Cells spherical, ellipsoidal cells that
contain abundant cytoplasmic
immunoglobulins and little to no surface
immunoglobulins
▫ The most fully differentiated lymphocyte and
their main function is antibody production
▫ Plasma cells are non-dividing, and after
several days of antibody production, they die
without further proliferation; (those that don't
die will become memory cells)
6. Memory
▫ Progeny cells of any stimulated B cells that
are capable of responding to antigen with
increased speed and intensity
CD19- present in all population of B cells
ADDED NOTES:
● Mature B cells will become Activated B cells if
it is exposed in an antigen; if NOT it will just
die thru APOPTOSIS.
● The immune antibody response begins when
individual B cells encounter an antigen and
they bind to the specific Immunoglobulin
surface receptors
● T cells are the ones that are being presented
to the antigen, there are 2 kinds of T cell: T
helper and T cytotoxic. The T helper are the
ones that send signals to the B cell for it to be
stimulated.
Stages of T Cell Differentiation
1. Pro-thymocyte
▫ Possess CD44 and TdT
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15
2. Double Negative Thymocyte (actively proliferates in
the outer cortex of thymus under the influence of IL-7,
very important in proliferation of DNT)
▫ Lack CD4 and CD8 antigens
▫ Possess CD2, CD5, CD7, CD45R
▫ Rearrangement of the genes that code for
the antigen receptor - T-cell receptor (TCR)
begins at this stage
3. Double Positive Thymocyte
▫ Express both Cd4 and CD8 antigens on their
surface as well as CD3-ab (TCR/ T cell
receptor)
▫ Would undergo Positive and Negative
selection
▪ POSITIVE SELECTION
▫ Allows only double-positive cells with
functional TCR receptors to survive.
▫ T cells must recognize foreign antigen in
association with class I or class II MHC
molecules→ present antigen to T cells
▫ MHC restriction (MHC found on the
endoplasmic reticulum of your phagocytes)
- The selection of thymocytes that will only
interact with the MHC antigens found on host
cells
▫ Any thymocytes that have either a very low
or a very high affinity for self-MHC antigens
die by apoptosis
ADDED NOTES:
● Thymocytes (T cells)- are Lymphocyte
precursors in thymus
● T cells (60-80% of circulating lymphocytes)
● Lymphocyte progenitor cells will be released
from the bone marrow and will enter the
Thymus → known as thymocytes → undergo
different gene arrangement and undergo
subsequent stages of differentiation
● When phagocyte engulf the antigen, it will be
digested and some of the digested peptides
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021
will be taken up by the MHC molecules which
is found inside the cells and it will be taken in
the surface of phagocyte → go to the T cell →
present it to the T cell → ACTIVATION of T
cell→ T helper cell will send signal to the B
cell so that it will be activated as well to
produce plasma cells→ antibodies
POSITIVE SELECTION
ADDED NOTES:
● Progenitor stem cells will exit the bone
▪ NEGATIVE SELECTION
▫ Takes place among the surviving DPT cells
▫ Strong reactions with self peptides other
than MHC antigens triggers apoptosis
ONLY ALLOW T CELL THAT DOES NOT REACT
NEGATIVELY TO SELF ANTIGEN
NEGATIVE SELECTION
4. Mature T cells (survivors of Selection Process)
T-CELL DEVELOPMENT
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16
marrow and will later on enter the thymus→
Prothymocytes→ Differentiation into Double
Negative Thymocytes through the
interaction/regulation of IL-7; LACKS CD4 AND
CD8 → will developed into Double Positive
Thymocytes; possess CD4 and CD8; consists
of 2 selection process which are Negative and
Positive Selection→ Positive Selection (all of
the FUNCTIONAL TCR receptors and cells that
can interact with MCH classes will SURVIVE,
those which are not will undergo Apoptosis →
Negative Selection (all T cells that produced
STRONG REACTION will die thru apoptosis;
ALL THAT SURVIVE will have either CD8 (T
cytotoxic) or CD4 (T helper) on its surface
▫ Represents those population of thymocytes
that had survived positive and negative
selection
▫ Express only 1 of either CD4 or CD8 on their
surface
▫ 2/3 CD4+ (T helper cell) → recognize
antigen that is presented by MHC Class 2
▫ 1/3 CD8+ (T cytotoxic cell) → recognize
antigen that is presented by MHC Class 1;
considered as “cell mediated immune
response”
▫ Subsets of Th cells:
• Th1 cells: interferon gamma (IFN-γ),
interleukin-2 (IL-2), and tumor
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
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necrosis factor-β (TNF-β) (Th1- T cell
activation)
• Th2 cells: IL-4, IL-5, IL-6, IL-9, IL-10,
and IL-13 (Th2- B cell activation to
become plasma cells and memory
cells)
- Help B cells produce antibodies
against extracellular pathogens
- Regulate B-cell activity
• T regulatory (Treg) cells: IL-10 - CD4
antigen as well as CD25 (Inhibit
proliferation of other T cell by
secreting inhibitory cytokines such as
IL-10, the response in antigen specific)
● T cells end product are CYTOKINES
Stages of T Cell Differentation
(Mature T Cells)
Other Subsets of Th cells:
▫ Th9: interleukin-9 (IL-9) and appear to have a
pro-inflammatory effect
→ IL-9: proinflammatory effect where it warns
extracellular fungi or bacteria
▫ Th17: Interleukin-17 (IL-17) and interleukin-22 (IL-
22)
→ IL-17: increased inflammatory response and joint
destruction associated with autoimmune response
such as arthritis, rheumatoid arthritis, multiple
sclerosis etc
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5. Activated T cells
▪ When antigen recognition occurs in the secondary
lymphoid tissue, T lymphocytes are activated and
differentiate into functionally active small
lymphocytes that produce
▫ cytokines Express receptors for IL-2 and
produce cytokines
▫ Cytokines functions include:
- Assisting B cells in the
commencement of antibody
production, (Th2 cytokines)
- killing of tumors and target cells
(CD8)
- rejection of grafts
- stimulation of hematopoiesis in the
bone marrow
- initiation of delayed hypersensitivity
allergic reactions.
▪ This type of immune response is known as
cell-mediated immunity.
6. T memory cells
Natural Killer Cells
▪ Ability to naturally kill or mediate cytolytic (lysing of
cell) or kill target cells without prior exposure to
antigen.
▪ Larger than T and B cells
▪ Contains a kidney shaped nucleus with condensed
chromatin and prominent nucleoli.
▪ Has high cytoplamsic: nuclear ratio
▪ Cytoplasm has many azurophilic granules
▪ No specific surface markers
▪ Possess CD16, CD56, CD94
▪ Lack CD3, CD4, and CD8
▪ Play a complementary role to CD8+ T cells
▪ Become LAK cells in response to IL-2
▪ Larger than T and B cells
▪ Contains a kidney shaped nucleus with condensed
chromatin and prominent nucleoli.
▪ Has high cytoplamsic: nuclear ratio
▪ Cytoplasm has many azurophilic granules
 IMMUNOLOGY AND SEROLOGY
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1st SEM, 2021

▪ No specific surface markers ▫ Fluorescent antibodies are used to screen of

▪ Possess CD16, CD56, CD94 subpopulation of T and B cells
▪ Lack CD3, CD4, and CD8
▪ Play a complementary role to CD8+ T cells
▪ Become LAK cells in response to IL-2 Components:

a. Sample delivery system

Mechanisms of NK Cell Cytotoxicity
1. It is brought about by the balance between b. A laser for cell illumination
activating and inhibitory signals that enables NK cells
c. Photodetectors for signal detection
to distinguish healthy cells form infected or cancerous
cells d. computer based management system
▫ Killer cell inhibitory receptors (KIRs) 4. Rosetting
▫ NKG2D: binds to MICA and MICB proteins 5. ELISA
on diseased or cancerous cells
SUMMARY
▫ If an inhibitory signal is not produced, NK
cells will release Perforins and Granymes
(hydrolytic enzymes → kill)

2. Antibody Dependent Cell Cytotoxicity

▫ Through binding of IgG-coated cell with

CD16 (receptor for fc function of IgG)

Laboratory Methods
1. Density Gradient Centrifugation with Ficoll Hypaque

2. Fluorescence Microscopy

a. Direct Immunofluorescence

▫ Use monoclonal antibodies with a

fluorescent tag (e.g. Fluorescein and
phytoerythrin- 490nm; rhodamine
545nm)

b. Indirect Immunofluorescence

▫ Uses unlabeled antibody that first

combines with the antigen by itself
and a second antibody that is
complexed with a dye

Laboratory Identification of Lymphocytes

3. Cell flow Cytometry/ Fluoresence Activated Cell
Sorter

▫ An automated system for identifying cells based on

the scattering of light as cells flow in single file
through a laser beam.

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021
The Nature of Antigens and the Major
Histocompatibility Complex
At the end of this unit, the student is able to:
▫ Differentiate antigen from an immunogen correctly.
▫ List and describe the different types of antigens
immunogen in terms of properties and
characteristics.
▫ Explain the role of adjuvants in enhancing immune
response
▫ Accurately discuss the general characteristics and
importance of the Major Histocompatibility Complex
▫ Differentiate and discuss the different classes of
HLA in terms of structure and function.
▫ Discuss the role of class I MHC and class II MHC
molecules in the presentation of antigens to T cells
▫ Identify the importance of HLA antigens in
transplantation.
▫ Enumerate and describe the types of grafts
correctly and identify the importance of graft
rejection in transplantation immunology
Part I: Nature of Antigens
Antigens
▫ A substance that reacts with antibody to sensitized
cells but may or may not provoke an immune
response
▫ An antigen is a substance that stimulates antibody
formation and has the ability to bind to an antibody.
▫ Foreign substance can be immunogenic or
antigenic.
Immunogen
▫ A macromlecule capable of eliciting the formation
of immunoglobulins or sensitized cells in an
immunocompetent host
▫ A foreign substance that induces a specific immune
response.
Factors affecting the Immune Response
1. Age
2. Dose
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19
3. Route of inoculation
4. Health status of the host
5. Genetics
Characteristics of an Immunogen
1. Foreignness
▫ the immune system normally discriminates
between self and non-self such as that only
foreign molecules are immunogenic.
2. Size
▫ However, in general, the larger the molecule
the more immunogenic it is likely to be.
3. Chemical composition
▫ In general, the more complex the substance
chemically the more immunogenic it will be.
▫ The antigenic determinants are created by
the primary sequence of residues in the
polymer and/or by the secondary, tertiary or
quarternary structure of the molecule.
4. Physical form
▫ In general particulate antigens are more
immunogenic than soluble ones and
denatured antigens more immunogenic than
the native form
5. Degradability
▫ Antigens that are easily phagocytosed are
generally more immunogenic. This is because
for the most antigens (T-dependent antigens)
the development of an immune response
requires that the antigen be phagocytosed,
processed and presented to helper T cells by
an antigen presenting cell (APC)
▫ APC – a heterogenous group of
immunocompetent cells that mediate the
cellular immune response by processing and
presenting antigens to the T-cells.
Chemical Nature of Immunogens
Proteins
▫ the vast majority of immunogens are
proteins. These may be pure proteins or they
may be glycoproteins or lipoproteins. In
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

general, proteiins are usually very good Epitopes on a bacterial cell

immunogens.
Polysaccharides
▫ pure polysaccharides and
lipopolysaccharides are good immunogens.
Nucleic acids
▫ are usually poorly immunogenic. However,
they may become immunogenic when single
stranded or when complexed with proteins.
Lipids
▫ in general lipids are non-immunogenic,
although they may be haptens
Epitopes and Haptens
1. Antigenic determinant/ Epitope
▫ Molecular shape or configurations that are
recognized by antibody or T cells
▫ May be either linear or conformational, or
may have repeating units or different
specificities
▫ that portion of an antigen that combines
with the products of a specific immune
response. It binds to the antigen-binding site
of antibody receptors.
HAPTEN
▫ Non-immunogenic materials that when combined
with a carrier create new antigenic determinants
▫ Haptens are small molecules which could never
induce an immune response when administered by
themselves but which can when coupled to a carrier
molecule.
2. Carrier
▫ Responsible to give the antigen its required
size
Types of Antigens
1. Autoantigens
▫ Antigens that belong to the host
2. Alloantigens
▫ Derived from the body of other individuals of the
same species and these are capable of eliciting an
immune response
3. Heteroantigens
▫ Derived from other species such as other animals,
plants, or microorganisms.
4. Heterophile antigens
▫ Those that exist in unrelated plants or animals but
which are either identical or closely related in
structure so that antibody to one will cross react with
antigen of the other
▫ Ex: polysaccharide type XIV of pneumococcus
reacting with anti-A antisera
Histocompatibility Antigens
▫ Referred to as the Human Leukocyte Antigen (HLA)
system in humans because its gene products were
originally identified on white blood cells
▫ These antigens are second only to the ABO antigens
in influencing the survival or graft rejection of
transplanted organs.
Blood Group Antigens
▫ In addition, certain antigens, especially those of the
Rh system, are integral structural components of the
erythrocyte (RBC) membrane.

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 IMMUNOLOGY AND SEROLOGY
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1st SEM, 2021

▫ If these antigens are missing, the erythrocyte

membrane is defective and results in haemolytic
anemia

CROSS REACTIVITY
▫ Denotes a situation in which two or more
compounds that may have various degree of
dissimilarity, share antigenic determinants/ epitope
and would therefore react with immune components
induced against any one of the compounds
Types of Antigens
T-Independent antigens Adjuvants
▫ They are usually carbohydrates which are composed ▫ A substance administered with an immunogen that
of monotonously repeating epitopes increases immune response against the immunogen

▫ They can directly stimulate the B cells to produce ▫ Substance that can enhance the immune response to
antibody without interacting with T cell. an immunogen are called adjuvants.

Properties of T-independent antigens ▫ The use of adjuvants, however, is often hampered by

▫ Polymeric structure – these antigens are
characterized by the same antigenic
determinant repeated many times.
▫ Polyclonal activation of B cells – many of
these antigens can activate B cell clones
specific for other antigens (polyclonal
activation)
T-dependent antigens
▫ Antigens which require the help of T-cells for the
formation of antibody
undesirable side effect such as fever and
inflammation.
▫ A pharmacological and/or immunological agent that
modifies the effect of other agents.
▫ Adjuvants are inorganic or organic chemicals,
macromolecules or entire cells of certain killed
bacteria, which enhance the immune response to an
antigen.
▫ They may be included in a vaccine to enhance the
recipient's immune response to the supplied antigen,
thus minimizing the amount of injected foreign
material.

▫ T dependent antigens are those that do not directly
stimulate the production of antibody without the help
of T cells.
▫ Proteins are T-dependent antigens. Structurally these
antigens are characterized by a few copies of many
different antigenic determinants.
▫ Most immunogens are thymic dependent
▫ Adjuvants are also used in the production of
antibodies from immunized animals. The most
commonly used adjuvants include aluminum
hydroxide and paraffin oil
Ways of Enhancing the Immune Response by
Adjuvants
1. It prolongs the existence of immunogen in the area
2. It increases the effective size of an immunogen
3. It increases the number of macrophages involved in
antigen processing

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

Location of the class I,II and III genes on

chromosome 6
Part II: The Immune Response
The Major Histocompatibility Complex
And HLA Typing
▫ The major histocompatibility complex (MHC) is a
genetic region, whose products play a role in
intracellular recognition and in discrimination
between self and non self.
▫ The MHC participates in the development of immune
responses, especially T-cell mediated immune
response.
Major Histocompatibility Complex (MHC)
Major Histocompatibility Complex (MHC) ▫ MHC molecules are actually found on surfaces of
▫ Large multi-gen locus consisting of several thousand cells in the body, and they play a pivotal role in the
kilobase pair of DNA on a single chromosome development of both humoral and cellular immunity.

▫ MHC complex forms a group of closely linked genes
that controls not only the exchange of tissues (tissue
compatibility) but also the myriads cellular interaction
of immune cells, the production of certain serum
proteins and the production of some cytokines and
enzymes
▫ MHC genes code for proteins that play a pivotal role
in immune recognition and it is thought that this
polymorphism is essential to our survival because it
allows for an immune response to diverse
immunogens
▫ In humans, it is referred to as Human Leukocyte
Antigen (HLA)
▫ Genes coding for the MHC molecules in humans are
found on the short arm of chromosome 6 and are
divided into three categories or classes:
▫ Class I molecules are coded for at three
different locations or loci, termed A, B, and C.
▫ Class II genes are situated in the D region,
and there are several different loci, known as
DR, DQ, and DP.
▫ For the class II molecules, there is a gene
that codes for the alpha chain and one or
more genes that code for the beta chain.
▫ Between the class I and class II regions on
chromosome 6 is the area of Class III genes,
which code for complement proteins and
cytokines such as tumor necrosis factor
▫ Their main function is to bring antigen to the cell
surface for recognition by T cells, because T-cell
activation will occur only when antigen is combined
with MHC molecules.
▫ Genes controlling expression of these molecules are
actually a system of genes known as the major
histocompatibility complex (MHC)
Structure of Molecules
Class I MHC molecules
▫ Comprised of two polypeptides:
▫ a polymorphic heavy chain (Alpha chain)
and a non-polymorphic light chain called B2
microglobulin.
▫ The heavy chain is encoded within the MHC
locus on chromosome 6, while the light
non-polymorphic chain is encoded on
chromosome 15.
▫ Each of the three forms of class I molecules:
▫ HLA-A, HLA-B, HLA-C are composed on
virtually all nucleated cells, although the
number of each HLA molecules (A, B and C)
expressed on any given cell varies.
Class II MHC molecules
▫ They are found primarily on antigen-presenting cells,
which include B lymphocytes, monocytes,
macrophages, dendritic cells, & thymic epithelium.

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 IMMUNOLOGY AND SEROLOGY
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▫ These molecules are comprised of two polypeptide
chains designated alpha and beta.
▫ Each of these chains is polymorphic and the genes
that encode then are located within the MHC locus.
▫ For the most part, class II MHC molecules HLA-DP,
HLA-DQ, HLA-DR are normally expressed on antigen
presenting cells
▫ Exposure to some cytokines can enhance the
number of class II MHC molecules expressed on an
antigen presenting cell.
▫ T cells can only “see” and respond to antigens when
they are combined with MHC molecules.
▫ It is thought that the two main classes of these
molecules have evolved to deal with two types of
infectious agents:
▫ those that attack cells from the outside
(such as bacteria)
▫ those that attack from the inside (viruses
and other intracellular pathogens).

▫ Additionally, some cells can be induced to express ▫ Class I molecules

these molecules de novo when activated ▫ Mainly present peptides that have been
synthesized within the cell to CD8 (cytotoxic)
T cells

▫ Class II molecules

▫ Present antigen to CD4 (helper) T cells

▫ Class I molecules are the watchdogs of viral, tumor,

and certain parasitic antigens that are synthesized
within the cell

▫ Class II molecules stimulate CD4 T cells in the case of

MHC Genetic Regions
bacterial infections or the presence of other material
▫ Class I region: HLA-A, HLA-B, and HLA-C
that is endocytosed by the cell
▫ Class II region: HLA-DP, HLA-DR and HLA-DC
Major Histocompatibility Complex (MHC)
▫ Class II region: complement proteins, cytochrome
p450, 21- hydroxylases and TNF
** Each of these genes exist in multiple different
allelic formation.
Natures of HLA Antigens
1. Glycoprotein component of cell membrane
2. Present in all surfaces of cells
3. Products of the genes of MHC
Roles of Class I & II Molecules in
The Immune Response
▫ The main role of the class I and class II MHC
molecules is to bind peptides within cells and
transport them to the plasma membrane, where T
cells can recognize them in the phenomemon known
▫ Class I molecules are synthesized in the rough
as antigen presentation.
endoplasmic reticulum, and for a time they remain
anchored in the endoplasmic reticulum membrane

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▫ Class II molecules are synthesized in the endoplasmic

reticulum and associate with a protein called the
invariant chain (Ii)

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 IMMUNOLOGY AND SEROLOGY
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Class I

(a) Antigen-processing pathway for endogenous

antigens

(b) The CD8+ T cell in association with class I MHC. The CD4+ T cell in association with class II MHC

Class II
The Role of the T Cells in the Adaptive Immune
Response
The Biology of Immune Response:
Antigen Presentation
Actions of T Helper Cells
▫ The first activating signal to induce transformation of
a T cell occurs when a CD4+ T cell encounters an
antigen along with a class II MHC molecule and binds
by using its antigen receptor.
▫ Within 1 to 2 days after antigen recognition has
occurred, T lymphocytes are transformed into large
activated blast cells that are characterized by
polyribosome-filled cytoplasm.

Antigen-processing pathway for exogenous antigens

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 IMMUNOLOGY AND SEROLOGY
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▫ They then begin cell division and generate progeny
with an antigen receptor that is identical to that of the
parent T cell.
▫ These functionally active small lymphocytes produce
cytokines
Actions of T Cytotoxic Cells
▫ Once activated by antigen in the lymph nodes or
spleen, cytotoxic T cells leave the secondary lymphoid
tissue and circulate to sites of infection.
▫ They bind and kill infected cells by triggering
apoptosis
▫ Act as a primary defense against intracellular
pathogens such as viruses, as well as other altered
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26
host cells such as tumor cells that exhibit new
antigens
▫ Cytotoxic T cells can kill target cells in two major
ways:
▫ Release the contents of granules that
damage the cell
▫ Bind to the host cell and, using intracellular
signaling, induce apoptosis.
▫ Granzymes
▫ a class of enzymes called serine proteases
▫ Perforins
▫ Pore-forming proteins that insert themselves
into the target cell membrane
▫ Once a cytotoxic T cell has bound to a target
cell, the granules move within the T cell to the
site where the target cell has bound
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

The Role of the B Cells in the Adaptive Immune

Response
The Biology of Immune Response:
Antigen Presentation
Response to T-Independent Antigens
B cells require Two signals to be activated
▫ The first occurs when antigen binds to membrane
immunoglobulin receptors on the B-cell surface and
cross-links them.
▫ The second signal is provided by Th cells themselves,
which bind to the B cell both through its antigen
receptor and through CD40 on the B cell and CD40L on
the activated Th cell.
▫ The bound T cell then delivers cytokines and other
signals to fully activate the B cell

Response to T-Independent Antigens

▫ T-independent antigens: antigens able to elicit
antibody formation in the absence of T cells.
▫ It only produce IgM because the induction of
memory cells does not occur to any great extent
▫ Several antigen receptors must be cross-linked in
order to activate a B cell directly MHC Genes and Diseases
▫ Epidemiological studies have shown that the HLA
alleles that an individual inherits may predispose that
individual to certain diseases.
▫ Individuals who inherit the HLA-B27 allele are at risk
for ankylosing spondylitis, an inflammatory disorder.
▫ Not all individuals who carry the HLA-B27 allele

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

develop ankylosing spondylitis; however, those with

the disease typically carry the HLA-B27 allele
HLA Typing
▫ Another inflammatory disorder, rheumatoid arthritis
occurs primarily in individuals who carry the HLA-DR4
allele.

How is HLA performed?

Application of HLA Typing/Matching
▫ Organ transplantation
▫ Paternity testing
▫ Forensic medicine, anthropology
▫ Studies of racial ancestry and migration
▫ For diagnostic and genetic counseling
▫ Basic research in immunology
AMethods for detecting HLA antigens:
Histocompatibility Testing
Tissue Typing
1. Serological approach (Lymphocyte microtoxicity
method)
▫ For determination of class I antigens
2. Cellular approach (Mixed Lymphocyte reaction)
▫ For determination of class II antigens
3. Molecular approach (PCR)
• A 20-30 ml blood sample is required to perform HLA
typing.
• The white cells are isolated from the blood.
Typing is performed by two different methods:
a. Serological testing: where the white cells
are used
b. DNA testing: where DNA extracted from
the white cells are used
-Preliminary tissue typing takes about 2 weeks.
-Further high resolution (more detailed) tissue typing
is performed on the patient and any potentially
matched donor samples may take another 2 to 4
weeks.
Types of Grafts
▫ Allograft
▫ Isograft
▫ Autograft
▫ Xenograft

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

Graft versus Host Disease

▫ The cells from the donor's immune system which are

introduced along with the transplanted stem cells
("graft") recognize HLA mismatches and attack vital
organs of the recipient's body ("host")

▫ T-cells from the transplant recognize the host MHC

molecules as non-self and attack the host

▫ Type IV HPS

Types and Tempo of Rejection

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

CYTOKINES - Responsible for many of the physical symptoms

• Small soluble proteins that regulate the immune
system, orchestrating both innate immunity and the
adaptive response to infection
• Chemical messengers that influence the activities of
other cells.
Actions of Cytokines
• Autocrine – affecting the same cell that secreted it
attributed to inflammation, such as fever, swelling,
pain, and cellular infiltrates into damaged tissues
Cytokines involved in the innate immunity:
• Non-immune (Type 1) Interferons
o IFN-α, IFN-β
• Tumor Necrosis factor

• Paracrine – affecting a target cell in close proximity • Interleukins

• Endocrine - systemic o IL-1, IL-6, IL-8, IL-10 , IL-12, IL-15

• Chemokines
Features of Cytokines
• Pleotropism - Single cytokine has many different - A family of cytokines that enhance motility
actions and promote migration of many types of
• Redundancy - Different cytokines often have very WBCs toward the chemokine source via a
similar effects process known as chemotaxis.

• Synergy - Cooperative effect of multiple cytokines - Most of the chemotactic activity of

• Antagonist - Inhibition of one cytokine effects by
another cytokine
• Act in networks - Stimulate the release of other
cytokines
• Act as growth factors for hematopoietic cells -
Modulate the number and composition of cells
leukocytes is regulated by the activities of
chemokines
- Classified into four families based on the
position of N-terminal cysteine residues: 
● C-X-C (alpha group), C-C (beta group), C
subgroup

Roles of Cytokines
• Innate immunity Functions of Cytokines in Innate Immunity
IL-1
• Adaptive immunity ● Induced by the presence of microbial pathogens,
bacterial lipopolysaccharides, or other cytokines
• Growth and differentiation of immature leukocytes
● Proinflammatory cytokines produced by
Cytokine Categories/Families monocytes, macrophages, and dendritic cells
• Tumor Necrosis Factor
● Induces fever, activation of phagocytes, and
• Chemokines
production of acute-phase proteins
• Interleukins
● Induce the production of vascular cell-adhesion
• Interferons molecules as well as chemokines and IL-6.

• Hematopoietic Factors ● Induces the production of CSFs in the bone

marrow, thereby increasing the available number
Cytokines in the Immune System of phagocytic cells that can respond to the
Cytokines in Innate Immunity damaged tissues

Tumor Necrosis Factors (TNF)

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

Induce lysis in tumor cells ● Acts as a control to help downregulate the

TNF-α
● Most prominent member of the TNF superfamily
● Exists in both membrane-bound and soluble
forms and causes vasodilation and increased
vasopermeability
● Production is triggered by the presence of
lipopolysaccharide, which is found in
gram-negative bacteria
IL-6
● Released in response to lipopolysaccharide and
plays an important role in acutephase reactions
● Expressed by a variety of normal and transformed
cells, including T cells, B cells, monocytes and
macrophages, vascular endothelial cells, and
various tumor cells.
● It affects inflammation, acute-phase reactions,
immunoglobulin synthesis, and the activation
states of B cells and T cells.
● Stimulates B cells to proliferate and differentiate
into plasma cells
inflammatory response when no longer needed
● Functions as both an activator and an inhibitor of
proliferation of affected cells
● Inhibits the activation of macrophages and the
growth of many different somatic cell types and
functions as an anti-inflammatory factor for
mature T cells.
● It blocks the production of IL-12 and strongly
inhibits the induction of IFN-gamma
Interferons (IFN)
Produced by dendritic cells and induce production of
proteins that directly interfere with viral replication
and cell division
IFN-α
Has been used to treat hepatitis C and Kaposi’s
sarcoma, as well as certain leukemias and lymphomas
IFN-β
Has been known as efficacious in treating multiple
sclerosis

● Induces CD4+ T cells to produce greater

quantities of both pro- and antiinflammatory
cytokines

Chemokines
● Chemotactic activities include: response to
infectious diseases, autoimmune inflammation,
cancer, and the homing of lymphocytes to all the
lymphoid tissues.

● Production is induced by both TNF-α and IL-6 Cytokines in Adaptive Immunity

● Facilitate the extravasation of leukocytes into the
tissues.
Transforming growth factor-β (TGF-β)
● Induces anti-proliferative activity in a wide variety
of cell types.
● A regulator of cell growth, differentiation,
apoptosis, migration, and the inflammatory
response
• Th1 cytokines - IL-2, IFN-γ, IL-12
• Th2 cytokines - IL-4, IL-5, IL-10
• Cytokines Associated with T Regulatory Cells (Treg
cells) - TGF-β, IL-10, IL-35
• Th17 - IL-17
Functions of Cytokines in Adaptive Immunity
Th1 Cytokines
IL-12

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

■ Produced by Dendritic cells in damaged tissues IL-13

■ Also produced by macrophages and B cells and
has multiple effects on both T cells and NK cells
■ Binds to its receptor on naïve T cells and causes
the expression of a new set of genes, including
those that determine maturation into the Th1
lineage
■ Increases the cytolytic ability of NK cells
■ Most properties are same as IL-4
■ Induce worm expulsion and favor IgE-class
switching
■ Plays an anti-inflammatory role by inhibiting
activation and cytokine secretion by monocytes
IL-10
■ Has inhibitory effects on the immune system.

Interferon-γ (IFN-γ) ■ Produced by monocytes, macrophages, CD8+ T

■ Principal molecule produced by Th1 cells cells, and Th2 CD4+ T cells

■ Stimulate antigen presentation by class I MHC ■ Has anti-inflammatory and suppressive effects on
and class II MHC molecules Th1 cells.

■ Most potent activator of macrophages and boosts ■ Inhibits antigen presentation by macrophages
their tumoricidal activity and dendritic cells

■ Stimulates the phagocytic and cytotoxic abilities ■ Inhibition of IFN-gamma production via the
creating activated or “super” macrophages. suppression of IL-12 synthesis by accessory cells

■ Involved in regulation and activation of CD4+ Th1 ■ Serves as an antagonist to IFN-gamma—it is a

cells, CD8+ cytotoxic lymphocytes, and NK cells. downregulator of the immune response

IL-2
■ Also known as the T-cell growth factor,

■ Drives the growth and differentiation of both T

and B cells and induces lytic activity in NK cells.

■ Together with IFN-γ induce the development of

Th1 cells, which, in turn, induces macrophage
activation and delayed type hypersensitivity

Th2 Cytokines
IL-4
■ One of the key cytokines regulating Th2 immune
activities and helps drive antibody responses

■ Expressed on lymphocytes and on numerous

nonhematopoietic cell types

■ Induces production of MHC-I, IL-4, IL-5, IL-13, and

the costimulatory molecules CD80 and CD86

■ Stimulates the production of IgG2a and IgE

■ Together with IL-5, drives the differentiation and

activation of eosinophils in both allergic immune
responses and response to parasitic infections

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 IMMUNOLOGY AND SEROLOGY
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1st SEM, 2021

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021
Antibody Structure and Function
Intended Learning Outcomes
● Differentiate and describe the five classes of
immunoglobulins in terms of structure,
properties and functions.
● Correctly identify the parts and functions of
the basic antibody structure.
● Explain the different theories of antibody
production
● Compare and contrast the primary and
secondary responses to antigen effectively.
● Accurately relate the influence of monoclonal
antibodies to current laboratory testing
practices
OVERVIEW
● Antibodies are specific glycoproteins referred
to as immunoglobulins.
● They can be found in blood plasma, and in
many body fluids such as tears, saliva, and
colostrum. (secretory globulins - igA)
● The primary function of an antibody in the
body's defenses is to combine with antigen,
which can lead to bacterial or viral
neutralization.
● They are considered to be the main humoral
element of the adaptive immune response
ADDED NOTES:
- B lymphocytes when stimulated by an
antigen, they undergo differentiation
- B lymphocytes in spleen and lymph node
when stimulated by helper T cells or T
independent antigen, they can directly
stimulate your B cells
- So when your B cells become activated or
stimulated by an antigen, they will undergo
differentiation and they become deform blast
(?) and they become plasma cells.
- and the end product of plasma cells is what
we call our antibodies which are aka
immunoglobulins
- Antibodies can also provide opsonization ,
complement activation, a lot of effector
functions aside binding antigen and
neutralization
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34
Immunoglobulins (Ig)
● Glycoprotein molecules which are produced
by plasma cells in response to an immunogen
and which function as antibodies.
● The Immunoglobulins derive their name from
the finding that when antibody-containing
serum is place in an electrical field, the
antibodies, which were responsible for
immunity, migrated with globular proteins.
● Immunoglobulins are divided into five major
classes on the basis of a part of the molecule
called the heavy chain:
○ IgG, IgM, IgA, IgD, and IgE (with Ig being the
abbreviation for immunoglobulin)
ADDED NOTES:
- Some of the early experiments that attempt
to determine the structure of the antibodies
or the immunoglobulins involved serum
protein electrophoresis
- Serum protein electrophoresis means using
electrical field so that you will know different
components or structures of antibodies will
be separated out
- Serum is placed on an agarose and then an
electrical current is applied to separate out
the proteins.
- If electrophoresis is carried out at the pH of
8.6, most of the serum proteins or the
immunoglobulins can be separated out on the
basis of size and their electrical charge
- Because of electrophoresis separation, there
were five distinct bonds that are obtained in
this manner
- Those distinct bonds determine the different
five major classes of your immunoglobulins or
antibodies as IgG, IgM, IgA, IgD, and IgE
ANTIBODIES
1. Fixation of complement - This results in lysis
of cells and release of biologically active
molecules
2. Binding to various cell types - Phagocytic
cells, lymphocytes, platelets, mast cells, and
basophils have receptors that bind
immunoglobulins.
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

○ This binding can activate the cells to perform some
function.
○ Some immunoglobulins also bind to receptors on
placental trophoblasts, which results in transfer of the
immunoglobulin across the placenta. As a result, the
transferred maternal antibodies provide immunity to
the fetus and newborn
cells whether extravascularly ( through the
mononuclear phagocytic system) or
intravascularly (complement proteins)
FC portion - bind (mediator) ; fragment crystallizable
FAB - bind (antigen) ; fragment antigen binding
Classification of Antibodies

● Each antibody has at least two identical sites Five classes of Antibodies:
that bind antigen: Antigen binding sites. ● IgG - predominant or most increased
(Fragment Antigen Binding Site/ fab) concentration in the blood
● Valence of an antibody: Number of antigen ● IgA
binding sites. Most are bivalent. ● IgM
● IgD
ADDED NOTES: ● IgE - least concentration in the blood
- The binding of your antibody to an antigen
has no direct biological effect rather the ADDED NOTES:
significant biological effects are a - arranged in decreading concentration in the
consequence of the secondary effector blood (IgG - pinaka madami; IgE - pinaka konti)
functions of our antibody
- Therefore, the antibodies upon binding will Classification of Antibodies
trigger secondary effector function (mediate a
variety of different effector functions such as A. According to its sedimentation constant
activation or fixation of the complement
pathway or binding to different various cell IMMUNO SEDIMENTATION MOLECULAR
types. GLOBULIN COEFFICCIENT WEIGHT
- If antibody is already bound to a specific (in Daltons)
antigen, it will trigger the activation of our C1
or complement system IgG 7s 150, 00
- If complement system is activated, the end
result is lysis of that particular antigen Serum IgA 7s 160,000
- Epitope - the portion of antigen that combines
antigen binding sites Secretory IgA 9s; 11s; 13s 170,000
Properties of an Antibody
IgM 19s 900,000
1. Protein in nature
2. With high molecular weight IgD 7s 180,000
3. Present in serum/ plasma, saliva, semen, CSF
and other body fluids IgE 7s 190,000

ADDED NOTES:
ADDED NOTES:
- Made out of protein - 82 to 96%
- Svedverg - unit represented by small ‘s’
- Made out of carbohydrates - 4 to 18%
- The larger the molecule, the farther it will
travel thus, it will have a larger sedimentation
GENERAL FUNCTIONS OF IMMUNOGLOBULINS
coefficient
1. Neutralize toxic substances (binding of antigen
to an antibody triggers a secondary effector
B. According to temperature at which they react
function = destruction/lysis)
best:
2. Facilitate phagocytosis and kill microbes
3. Combine with antigens on cellular surfaces
and hereby cause the destruction of these
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 IMMUNOLOGY AND SEROLOGY
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1st SEM, 2021
1. Cold Antibodies - react the best at 4 to 22C or
room temperature (eg. IgM)
2. Warm Antibodies - react best using 37C (eg.
IgG)
- ABO blood group - mostly of type IgM
- RH blood group - IgG
- Anti-D - warmer acting antibodies and Anti-A
and Anti-B are cold reacting antibodies
C. According to occurrence
1. Natural antibodies - present at birth (eg. IgA)
2. Immune antibodies - are produced only after
antigenic stimulation
D. According to the species which produce them
1. Isoantibodies - produced from the same
specie; produced upon exposure to
alloantigen
2. Heterophile antibodies - produced from
different specie; produced upon exposure to
heterophile antigen
E. According to its reaction with an antigen
1. Agglutinins
- antibody; can be from IgG, IgA, IgM,
IgD and IgE
- involved in agglutination reaction;
- agglutinate upon binding to a specific
antigen;
- they form large complexes;
- reacts with antigen called
agglutinogen;
- particulate antigen result or cause
agglutination reaction
2. Precipitins
- antibodies that are able to react with
soluble antigen
- capable of precipitation reaction
- soluble antigen once bind to an
antibody cannot agglutinate; it will
only precipitate
- antigen: precipitinogen
3. Agglutinoids
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36
- modified agglutinins
- uses heat
- antigen: agglutinogen
- are not capable of agglutination
reaction and don’t form large
complexes
4. Hemagglutinins
- agglutinins using RBCs
- involved in agglutination of RBCs
- binds to antigens which are found in
the RBC such as anti antigen A and
antigen B
- once anti-A is bound to antigen A, it
will form agglutination reaction
- Anti-A and Anti-B are also known as
hemagglutinins
5. Lysins
- involved in the solution of lysis of
antigenic cells (eg. bacterialysin,
hemolysin)
- eg: ASO (anti-streptolysin O antibody)
if bound to RBC (lyses rbc)
6. Opsonins
- very good opsonin: IgG
- phagocytosis
7. Neutralizing antibodies
- or protective antibodies
- can make microorganisms harmless
- eg: ASO (anti-streptolysin O antibody)
if bound to SO antigen
8. Allergic antibodies
- allergic reaction
- IgE involved
9. Antitoxins
- neutralize specific toxins
10. Complement fixing antibodies
- implement fixation
11. Blocking or Inhibitory antibodies
- antibodies can block or inhibit other
antibodies or a particular reaction
 IMMUNOLOGY AND SEROLOGY
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1st SEM, 2021

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

F. According to their in vitro behavior ultracentrifuge and he separated out the

different structure of immunoglobulins on the
basis of the molecular weight while Rodney
Complete Incomplete
Porter worked on using proteolytic enzymes
Antibody Antibody
such as papine to know the different
(IgM) (IgG)
structures of an antibody
Synonyms Bivalent; Saline Univalent;
acting blocking; Tetrapeptide made up of:
-two binding coagglutinating 1. Heavy chain
site -needs another - With five principal antigenic types with their
antibody to corresponding Ig. (gamma, alpha, mu, delta,
agglutinate epsilon)
2. Light chain
Response to Thermolabile Thermostable - With two antigenically defined types: kappa
Temperature -cold temp -warm temp and lambda

Ability to cross Cannot cross Can cross the Two heavy chains and two light chains are all linked by
placenta the placenta placenta disulfide bonds that’s why they become one
-only IgG can monomer
cross the
placenta Antibody Structure
A four-chain unit molecule, each L chain was bonded
Occurence Early in Late in to an H chain by means of an S–S bond and the H
immunization immunization chains were joined to each other by one or more S–S
-primary; no -secondary bonds
memory response
Disulfide bonds
Reaction Saline acting Albumin acting These are chemical bonds essential for the normal 3
dimensional structure of Ig
ADDED NOTES: 2 types of disulfide bonds:
- Complete antibody - IgM 1. Interchain - disulfide bonds that connect parts
- Incomplete antibody - IgG of different chain (light chain and heavy
- Complete antibody is involved in primary chain)
response 2. Intrachain - connect parts within the same
- Complete antibody is produced upon chain
secondary response or anamnestic response

The Structure of Immunoglobulin

(The Tetrapeptide Structure of Immunoglobulins)

The Structure of Immunoglobulin

Monomer
- basic structural unit of an antibody
- made up of tetrapeptides or 4 protein
molecule
- two light chains, two heavy chains =
tetrapeptide
- basic structure of an immunoglobulin was first ADDED NOTES:
discovered by Gerald Edelman using
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 IMMUNOLOGY AND SEROLOGY
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1st SEM, 2021
- The exact number of the sulfide bonds differ
from antibody classes and subclasses
- FAB - fragment antigen binding
- FAB - has capacity to bind antigen; in portion
of antigenic determinant or epitope
- Once FAB binds to epitope, secondary effector
will initiate, which our whole cell will bind to
fc region
- Each FAB fragment is consist of one light chain
and one half of heavy chain which is held
together by interchain disulfide bonds
- FC - fragment crystallizable; crystalize at 4C
FC - important in effector functions of our
antibody molecules which can include
opsonization and complement fixation
- Host cells - bind in FC; can be our complement
proteins or our phagocytic or mononuclear
phagocytic system that can initiate
opsonization or complement fixation that
result to destruction of our antigen
Hinge region
● Flexible part of the antibody located in the
heavy chains.
● Located between the CH1 and CH2 regions is
known as the hinge region
● It is more exposed to enzymes and chemical
thus papain acts here to produce Fab and Fc
portions
● The light chains are each linked to one-half of
a heavy chain by disulphide bonds at the
proximal end
ADDED NOTES:
- flexibility because of its high proline content
- flexibility allows antibody to bend the 2
antigen binding site and operate
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39
independently and engage in angular motion
relative to the other knob based on its binding
capacity
- hinge connects the heavy chains that forms
“Y”
- Mu / IgM and Epsilon/IgE don’t have hinge
region
Light Chains
● Analysis of several Bence Jones proteins
revealed that there were two main types of L
chains, which were designated as:
○ kappa (κ) chains and lambda (λ) chains
An antibody molecule is composed of two identical Ig
heavy chains (H) and two identical light chains (L),
each with a variable region (V) & constant region (C).
ADDED NOTES:
- Each light chain contained between 200 to
220 amino acids and each type of chain had
essentially the same sequence
- Two regions on each light chain; we have the
carboxyl terminal end, which is the constant
region and the amino terminal end, which is
the variable region
- No functional differences between the two
types of chains. The differences lies in the
amino acid substitution along the chains
- Both kappa and lambda chains are found in all
5 classes of immunoglobulin but only 1 type is
present in a given molecule
- Constant heavy chain 2 and constant heavy
chain 3 makes up the FC region
- Variable heavy chain and light chain comprises
FAB region
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

Three-Dimensional Structure of Antibodies

● The basic four-chain structure of all

Heavy Chains
immunoglobulin molecules does not actually
exist as a straight Y shape.
● The amino-terminal end constitute the
variable domain; the remaining amino acids
● It is in fact, folded into compact globular
can typically be divided up into three or more
subunits based on the formation of
constant regions with very similar sequences,
balloonshaped loops at each of the domains
designated CH1, CH2, and CH3.

● Constant regions of the H chain are unique to

each class and give each immunoglobulin type
its name.

DOMAINS

● Are globular regions on polypeptide chain

stabilized by intrachain disulfide bonds
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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

a. Domains on the heavy chain DOMAINS

○ VH, CH1, CH2, CH3, (CH4)
Cleavage with papain enzymes
b. Domains in the light chain ○ Digestion with papain breaks the
○ VL, CL immunoglobulin molecule in the hinge region
before the H-H inter-chain disulfide bond.

○ This results in the formation of two identical

fragments that contain the light chain and the
VH and CH1 domains of the heavy chain.

○ Antigen binding - These fragments were

called the Fab fragments because they
contained the antigen binding sites of the
antibody.

Cleavage with papain enzymes

Regions on Polypeptide Chain

Cleavage with papain enzymes
Variable Region Constant Region
● Digestion with papain also produces a
Amino acid sequence Amino acid sequence is fragment that contains the remainder of the
subject to change fixed and unchanging two heavy chains each containing a CH2 and
CH3 domain. This fragment was called Fc
Amino terminal end Carboxyl terminal end because it was easily crystallized
(NH2) (COOH)
● Effector functions - The effector functions of
Concerned with binding Concerned with binding
immunoglobulins are mediated by this part of
to antigen to host tissue
the molecule.

Other Definitions Enzyme Digestion

Cleavage with pepsin enzymes
Polymer ○ Treatment of immunoglobulins with pepsin
○ Ig composed of more than a single basic results in cleavage of the heavy chain after the
monomeric unit H-H inter-chain disulfide bonds resulting in a
J chain fragment that contains both antigen binding
○ Polypeptide chain which normally holds sites
polymeric Immunoglobulins ○ This fragment was called F(ab')2 because it
Secretory component is divalent.
○ A substance attached to polymeric IgA ○ The Fc region of the molecule is digested
found on secretions. into small peptides by pepsin.
○ The F(ab')2 binds antigen but it does not
Enzyme Digestion mediate the effector functions of antibodies
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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

Reduction using mercaptoethylamine

○ Break the disulfide bonds

Cleavage with pepsin enzymes

Characteristics of the Different Types of

Immunoglobulins

Enzyme Digestion

Immunoglobulin Classes

IgG
● Structure: Monomer
● Percentage serum antibodies: 75- 75% of the
total Immunoglobulin pool
● Has 4 major subclasses: IgG1, IgG2, IgG3, IgG4
● Equally distributed in the different fluid
compartments with detectable amounts in
CSF and urine
● IgG antibody response appears later than IgM
in primary response but they form the major
antibody of the secondary immune response

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

IgM Response

● Maternal IgG is actively and selectively

transferred across the placenta to the fetus
and imparts passive protection to the newbon
for 6-9 months
Functions of IgG:
○ Provides immunity for ,the newborn
○ Complement fixation ○ Opsonization
○ Neutralization of toxins and viruses
○ Participation in agglutination and precipitation
reactions

IgM
● Structure: Pentamer
● Percentage serum antibodies: 5-10% of the
total immunoglobulin pool
● Star-shaped in the free state; reaction with 10
functional binding sites.
● crab-like in antigen-antibody Location: Blood
and lymph (pentamer), B cell surface
(monomer)
● Half-life in serum: 6 days
● A powerful agglutinator of a particulate
antigen
● Maternal IgM does not cross the placenta
IgA
● Structure: Dimer
● Percentage serum antibodies: 10-15% of
human serum Ig pool
● Found in serum in small amounts but
predominant in sero-mucous secretions of the
respiratory tract, genito-urinary tract and GI
tracts
● Secretions: (tears, sweat, colostrum, saliva
and breastmilk), blood and lymph.
● Half-life in serum: 6 days
● Complement Fixation: No
● Placental Transfer: No

Functions of IgM:
○ First antibodies produced during an infection but it
does not persist for long.
○ Complement fixation ● Forms of IgA:
○ Agglutination Serum IgA
○ Opsonization - Can agglutinate motile infectious agents thus
○ Neutralization of toxins promoting their phagocytosis but they cannot activate
○ Surface receptor for antigens (on B cells) the complement system

IgM Response Secretory IgA

- A polymeric form stabilized a short polypeptide
chain.

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

- It is known as the “antiseptic paint” of mucous
membranes.
- It can activate the bacteriolytic activtiy through the
alternate pathway of the complement system and only
in the presence of lysozyme
● It is homocytropic due to its ability to attach
to the human skin, it is associated with
immediate hypersensitivity reactions but also,
apparently, with immunity to certain
helminthic parasites

Known Functions: Localized protection of mucosal

surfaces. Provides immunity to infant digestive tract.

IgD
● Structure: Monomer
● Percentage serum antibodies: 0.001% of total
Ig pool, but known to be present in large
quantities on the membrane of many
circulating immunocompetent B lymphocytes
and unstimulated B cells
● Heat labile immunoglobulin
● Detected by highly sensitive assay requiring
radio-labeled antisera
● Precise biological action is not known but it
may play a role in antigen-triggered
lymphocyte differentiation
● Location: B-cell surface, blood, and lymph
● Half-life in serum: 3 days
● Complement Fixation: No
● Placental Transfer: No ● Also known as reaginic antibody/ nuissance
● Known Functions: In serum function is antibody
unknown. On B cell surface, initiate immune ● Location: Bound to mast cells and basophils
response throughout body. Blood.
● Half-life in serum: 2 days
● Complement Fixation: No
● Placental Transfer: No
Known Functions:
○ Allergic reactions. Possibly lysis of worms.

IgE
● Structure: Monomer
● Percentage serum antibodies: 0.0005% of
total serum Ig Theories of Antibody Production
● Most heat labile immunoglobulin
● Synthesized locally by plasma cell present in Ehrlich’s Side Chain Theory
the mucous membrane of the GI and ○ Certain cells has had specific surface
respiratory tracts. receptors for antigen that were present before
contact with antigen occurred.

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

○ Once antigens was introduced, it would

select the cell with proper receptors,
combination would take place and then
receptors will break off and enter the
circulation as antibody molecules.

○ New receptors will be formed in place of

those broken off and this process could be
repeated

Clonal Selection Theory

○ Individual lymphocytes are gentically
pre-programmed to produce one type of
immunoglobulin, and that specific antigen
finds or selects those particular cells capable
of responding to it, causing these to
proliferate

○ Repeated contact with the antigen would

continually increase a lymphocyte pool

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021
THE COMPLEMENT SYSTEM
Learning Intended Outcomes
● Explain the mechanism involved in the
classical pathway and the alterative pathway
of complement activation efficiently.
● Describe the roles of the complement system
in mediating immune response.
● Outline the process of complement activation
using the different pathways correctly.
● Correlate the complement factors based on
their role in the process of inflammatory
reactions.
● Correctly describe the principles of reaction
involved in the measurement of complement
components.
● Analyze laboratory findings and identify
disease implications in relation to
complement abnormalities
Overview
● Complement Protein - is a SERIES or complex
series of more than 30 heat labile proteins
that play a major role in INFLAMMATORY
RESPONSE or act a → Pro-inflammatory
mechanism of immune response
● The complement system is a heat labile
SERIES of PLASMA PROTEINS major role in the
immune system is → CYTOLYSIS as a
PRO-INFLAMMATORY RESPONSE or POTENT
MEDIATOR OF INFLAMMATION of
inflammation. many of which are enzymes or
proteinases.
● Complement plays a major role in the immune
system (Cytolysis) as a potent mediator of
inflammation (Pro-inflammatory).
● Activation of the complement system - will
lead to LYSIS of the antigen as a secondary
effector molecule of our antibodies
● A complement proteins can also serve as
Pro-inflammatory response - has the ability
also to act as anaphylatoxin, as a chemotaxin,
or as an opsonin.
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46
● Anaphylatoxin - wherein some of the
complement proteins have the ability to
INCREASE vascular permeability or to BIND to
muscles and basophils causing the granulation
and releasing of the vaso-active mediators
such as histamine, leukotrienes, and other
vaso-active mediators
● Complement proteins → can also serve as
chemotactic factors or could act as a
chemotaxin wherein it initiates a recruitment
of monocytes or phagocytes, neutrophils to
the area of antigen concentration, they
initiate CHEMOTAXIS.
● Complement proteins → can enhance
phagocytosis by acting as an opsonin; they
opsonize and tag invaders for clearance and to
facilitate phagocytosis and also to DIRECT the
adaptive immune system to the site of
infection
● Pro-inflammatory response → of
complement proteins serve as an IMPORTANT
LINK between innate and adaptive immunity
● Majority of the function of your complement
proteins is part of the INNATE while as a
secondary effector function of antibodies, it
also functions in the ADAPTIVE immunity
● Normally, complement components are
present in the circulation in an inactive form.
and when an antibody is combined to an
antigen, it will trigger the activation of the
complement system and the ultimate goal or
the end result of the activation of the
complement system is lysis of the antigen that
is bound to the antibody.
● Ultimate goal which is → CYTOLYSIS of
antigen. During the activation process, there
are also complement proteins that can be
activated and have a pro-inflammatory
response or mediates or a potent mediator of
inflammation.
● Aside on lysis, Complement proteins has
Pro-inflammatory role links the complement
system to our innate immunity.
 IMMUNOLOGY AND SEROLOGY
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● Complement is composed of three
interrelated enzyme cascades:
➔ Classical pathway
➔ alternative pathway
➔ mannose lectin binding pathways.
➔ ●
● Present in NORMAL SERUM but as part of the
innate immunity, complement can also be
considered as an ACUTE PHASE REACTANT
because the levels of our complement
proteins will RISE because of activation during
an infection. COMPLEMENT PROTEIN levels
will rise during an INFECTION.
● Complement protein has a UNIQUE property
of being EASILY inactivates as well; inactive
during circulation but when there is an
ANTIGEN that is BOUND TO ANTIBODY or
meron infection it RISES LEVEL and
complement proteins will ACTIVATE once
activated can have now the PRO
INFLAMMATORY ROLE which is anaphylatoxin,
chemotaxis, opsonin and lysis.
● BUT also Complement protein has UNIQUE
property also of being EASILY INACTIVATED
and Complement protein are VERY HEAT
LABILE and can be INACTIVATED by heating
the serum to 56C for 30 minutes - therefore
ma prevent na ang lysis and pro-inflammatory
role of complement protein.
Definition of terms:
C-activation
● Alteration of a complement (protein) in such a
way that it can proceed to interact with the
next component in the pathway (cascade).
● Complement system is a CASCADE meaning;
→ Activation of one will lead to an activation
of the other protein.
→ Absence of one complement protein will
PREVENT ACTIVATION OF OTHER therefore
(hindi magtuloy-tuloy yung formation, di
magkakaroon ng LYISIS since cascade)
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47
C-fixation
● Utilization of complement components by the
antigen antibody complex.
We only have 2 types of immunoglobulins that
can fix your complement proteins in their FC
portion → IgG & IgM
● BEST immunoglobulin that can FIX the
complement → IgM (because IgM is a
pentamer and it has a lot of binding sites)
● After IgM is IgG3; EXCEPT → IgG4
● IgG 4 → CANNOT FIX the complement as well
as IgA, IgE and IgD
C-inactivation
● Complement proteins are HEAT LABILE and
can easily inactivated by HEATING the SERUM
at 56C degree celsius for 30 minutes
● denaturation (usually by HEAT) of one of the
early components in –activation pathway
resulting in the destruction of C-hemolytic
activity.
Convertase/esterase
● ACTIVATED (altered/cleaved) COMPLEMENT
COMPONENT which act as a proteolytic
enzyme specific for subsequent components.
● C3 convertase → it converts or cleaves C3
complement proteins
● C5 convertase → act as PROTEOLYTIC ENZYME
that specific to cleave or to act upon C5
Proteins of the Complement System
● Most of the PLASMA complement proteins are
synthesized in LIVER or in the HEPATOCYTES
also macrophages and gut epithelial cells.
● Complement system is composed of more
than 25 different proteins produced by
different tissues and cells including
hepatocytes, macrophages and gut epithelial
cells.
● These proteins are ACTIVATED by a variety of
agents and their activation proceeds via
 IMMUNOLOGY AND SEROLOGY
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different pathways and if the activated
products bind to a cellular target, their
deposition leads to CELL LYSIS
● The ultimate goal or ultimate end result of
activation of the complement system is →
CELL LYSIS of the antigen but along the way
there are several complement components
that are ACTIVATED that can also can or have a
PRO-INFLAMMATORY ROLE in our innate
immunity.
● Most plasma complements are synthesized in
LIVER, but with the EXCEPTION C1
complement components are synthesized in
intestinal epithelial cells or gut epithelial cells
● Factor D - involved in the alternative system is
made or synthesized in → ADIPOSE TISSUE
● Other cells also synthesize other complement
components such as -
Macrophage, monocyte → additional sources
of C1, C2, C3 and C4 but MAJORITY is
produced in LIVER
● Complement components - is a CASCADE
that's why ACTIVATED
● Since the complement components are
activated in a CASCADE FASHION, the absence
of one of the components in the pathway CAN
DISRUPT the cascade and TERMINATE the
reaction.
● Remember: Activation of ONE complement
protein will LEAD → to ANOTHER activation of
a complement protein then to another until it
reaches the membrane attack complex which
will cause the LYSIS of the antigen.
● Absence of ONE component that is needed for
activation in the complement pathway can
DISRUPT the CASCADE and terminate
reactions
● Prone to infection - those who have
deficiencies in complement components
(Malaki ang contribution ng atin Complement
system in our Immune response)
COMPLEMENT SYSTEMS:
1. Classical pathway
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48
- which involves NINE proteins (C1 to C9) that
are triggered by →ANTIGEN ANTIBODY
COMBINATION
- activated when an antibody is BOUND to an
antigen (so kung di ANTIBODY meron
ANTIGEN na naka bound sakanya →it will
trigger the activation or our CLASSICAL
PATHWAY that’s why classical pathway is →
Antibody dependent pathway
2. Alternative pathway
- Originally called the PROPERDIN SYSTEM,
because the protein Properdin was thought to
initiate this pathway.
- Hindi antigen antibody combination ang nag
triggered ng activation ne to but → the
Lipopolysaccharides of the bacterial cell wall
ang nag activate ng Alternative pathway
Properdin - it JUST used to STABILIZE A KEY ENZYME
COMPONENT/COMPLEX that is formed along the
pathway during the alternative which is the → C3
convertase
Properdin - is know as ALTERNATIVE PATHWAY,
because it by passes C1 to C4 of classical
3. Lectin pathway
- is another antibody-independent means of
activating complement proteins.
- LIKE alternative because the
lipopolysaccharide of the bacterial cell wall
activates the activation of alternative
pathway while lectin pathway is activated by
→ MANNOSE which are primarily found in
fungi, bacteria, viruses, yeast, and
parasite/protozoa
MBL (Mannose binding lectin) - mediator of
LECTIN pathway becoz the mannose binding
lectin is the one mediate/initiates the LECTIN
PATHWAY
when the MBL it bound to the mannose will
activate the → LECTIN PATHWAY and the
ultimate goal is to LYSIS of the antigen.
➔ Its major constituent, mannose- (or
mannan-) binding lectin (MBL),
adheres to mannose found mainly in
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

the cell walls or outer coating of

bacteria, viruses, yeast, and protozoa.
➔ While each of these pathways will be
considered separately, activation
seldom involves only one pathway
● The first stage involves C1, which is known as
the recognition unit.
● Once C1 is fixed, the next components
activated are C4, C2, and C3, known
collectively as the activation unit.

● C5 through C9 comprise the membrane

attack complex, and it is this last unit that
Pathways of the Complement System completes lysis of the foreign particle
The Classical Pathway, The Alternative Pathway & The Recognition Unit
The Lectin Pathway
Review:

Jules Bordet - was AWARDED a Nobel Prize in 1919 for

his role in elucidating the nature of complement
1890 - complement was originally recognized
→ by Paul Erlich
Paul Erlich - one who COINED the term complement
because: COMPLIMENT the ACTION of the antibody in
DESTROYING microorganism.

Antibody - not the one that neutralizes the antigens

Secondary effector function that binds to FC portion -
neutralizes antigens; could be:
➔ intravascularly through LYSIS of the antigen is
through the action of complement proteins
➔ extravascularly if the monocytes or
macrophages will be the one to bind on the FC
portion (Mononuclear phagocytic system)
FIRST COMPLEMENT COMPONENT to bind → C1
CLASSICAL PATHWAY
● C1q “recognizes” the fragment crystallizable
● Classical pathway - is the MAIN antibody
dependent/ antibody directed. (FC) region of two adjacent antibody
molecules, and at least two of the globular
● NOT all immunoglobulins are able to activate heads of Clq must be bound to initiate the
the classical pathway nor the complement classical pathway.
system ● It must attach to an antigen within 30-40
● The only immunoglobulins that can activate nanometers → before complement can bind,
classical pathway are IgM (BEST), IgG1, IgG2 so it needs 2 ADJACENT ANTIBODY for a
and IgG3 compliment protein to BIND
➔ C1 - FIRST complement to bind;
● IgM is the BEST becos it’s PENTAMER it has known as → RECOGNITION UNIT
MULTIPLE BINDING SITE ➔
IgM → IgG3 →IgG1 →IgG2 ● C1 has three (3) subunits
➢ C1q
EXCEPT → IgG4 ➢ C1r
● IgG4 cannot fixed complement as well IgA, ➢ C1s
IgE, IgD
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● These 3 are stabilized by → CALCIUM
→ as long as CALCIUM is present in the
SERUM → these three (C1q, C1r, C1s) will
REMAIN associated.
● C1s & c1r REMAIN associated with → c1q
● ●
● This structure has been often referred to as a
BOUQUET OF TULIPS → with six blossoms
extending outwards
● INACTIVE in SERUM → but when ANTIBODY
binds to an ANTIGEN → C1q will RECOGNIZE
the FC portion
Remember:
IgM → CH3
IgG → CH2
● C1r and C1s - are both SERINE PROTEASE and
PRO ENZYMES or called →ZYMOGENS
●
● C1q - it is the part that binds to the FC portion
of antibody molecule
● C1r & C1s - subunits also generate enzyme
activity to begin the → CASCADE
● C1q, C1r, and C1s are needed to begin the
cascade
● As binding occurs at the CH2 region for →IgG
and at the CH3 region for →IgM
→ Both C1s and C1r are both converted into active
enzymes
● Mechanical stress transmitted from the stems
as binding occurs open up to activate → C1r
and C1s
→ Once C1r is activated, it will CLEAVE the thioester
band on → C1s
Activated C1r is extremely specific, because its only
known substrate is → C1s.
C1r → has a LIMITED SPECIFICITY being only → C1s
● Likewise, C1s has a limited specificity, with its
only substrates being → C4 and C2.
● → C2
● Once C1s is activated, the recognition stage
ENDS and the activation stage → BEGINS
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50
Activation stage BEGINS when → C1 cleaves yung
kanyan 2 only sunbrates which is → C4 and C2
The Activation Unit
C4 - second most abundant complement protein
The activation unit, BEGINS when
C1s cleaves → C4 into → C4a and C4b ends with the
production of the enzyme C5 convertase
● Once we have the production of C5
convertase, the 3rd stage of activation which
is the membrane attack complex will begin.
● C4 - SECOND most abundant complement
protein
C1s will cleave → C4 and split it into → C4a and C4b
C4b→ in the process, it OPENS the → THIOESTER
active site on the remaining part of the C4b → C4b
protein must bind to → C2 (para di ma degraded)
Once the C4b cleaved the next → component to be
activated is → C2
● C4b must bind within a FEW SECONDS to → C2
for it not to be DEGRADED by →
water/carbohydrate
● when combine with → C4b in the present of
MAGNESIUM IONS → C2 cleaves by also C1s
into → C2a & C2b
→ C2b will enter circulation → C2a will be used up in
the pathway
● C3 - the MAJOR and central constituent of the
ALL complement systems (present sa lahat ng
complement system)
● C3 → It serves as the PIVOTAL POINT for all
three pathways.
● The cleavage of C3 to C3b represents the
most significant step in the entire process of
complement activation
● C3b needed in the formation of enzymes such
as the C3 convertase & C5 convertase
● If C3b is bound to C4b2a, this creates a new
enzyme known as C5 convertase
ACTIVATION STARTS → C1s cleaves → C4a (enter
circulation acts as ANAPHYLATOXIN) & C4b combine
after forming C4b
C1s cleaves → C2 to → C2a & C2b
 IMMUNOLOGY AND SEROLOGY
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C4b will be used up and bind → C2a forming → C4b2A

known as C3 convertase
gagamitin sa Classical pathway → C4b & C2a

C3 convertase cleaves → C3a & C3b

If binding does occur
C3 cleaved into → C3a (will enter the circulation and
acts as an ANAPHYLATOXIN)
& C3b will join → C3 convertase
forming → C4b2A3b know as C5 convertase
C5 convertase - is NEW ENZYMES that
cleaves → C5a and C5b
C5b will be deposited/attach onto the cell membrane
and it will now BEGIN the formation of the →
MEMBRANE ATTACK COMPLEX
In the formation of membrane attack complex:
C3b - also serves as a POWERFUL OPSONIN and a
major contribution to the process of PHAGOCYTOSIS,
→ some C3b that has been activated can also go to
the circulation and act as OPSONIN.
ACTIVATION UNIT ends → C5 convertase (C5b)

The Membrane Attack Complex (MAC)

● Starts when C5 convertase, consisting of

C4b2a3b, splits a piece known as → C5a that
is released into circulation, whereas C5b
attaches to the cell membrane, forming the
beginning of the → MAC.
C5b → is EXTREMELY LABILE and rapidly inactivate
→ unless the C6 binds or bound
● Once C6 is bound to → C5b, subsequent
binding involves → C7, C8, and C9

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C5b6789 → known as MEMBRANE ATTACK COMPLEX

● The complex of C5b-C6-C7-C8 and C9 is known
as C5b-9 or MAC

These complexes attached to the surface → causing

the LYSIS
C6 → C7 bind next forming → TRIMOLECULAR
COMPLEX that has a HIGH AFFINITY for LIPID
CONSTITUENTS of the cell membrane → C8 - binding
exposes the HYDROPHOBIC REGION that interacts
with membrane to form a → SMALL HOLE
binding of C8 will cause → PORE FORMATION of cell
membrane of antigen
C8 will be inserted into → BILAYER OF THE LIPID and
→ LYSIS can be absorb at this stage
Membrane Attack Complex
C9 components - accelerates the process of LYSIS
The LYSIS or Membrane damage is caused by at least
→ So binding of C8 starts LYSIS & will cause → LOSS of two different mechanisms:
potassium, leakage of amino acid and
➔ Channel formation
ribonucleotide.
➔ the binding of phospholipids
but the binding and addition of MULTIPLE MOLECULES
of → C9 components → will accelerate this process
and → potentiate the LYSIS of the cell membrane. ● The latter binding of phospholipids causes a
→ reordering and reorientation of molecules
that results in → leaky patches (creates pore)
So, it need multiple C9 molecules to POLYMERIZE the
TRANSMEMBRANE CHANNEL and cause → LYSIS OF
THE CELL ● When complement proteins are bound,
membrane phospholipids rearrange
C8 - starts the LYSIS
themselves into domains surrounding the
Destruction of TARGET CELL → occurs through the → C5b6789 complex or MAC,, and the integrity
INFLUX or paglabas ng leakage of water and of the membrane is destroyed.
corresponding loss of electrolytes ●
SMALL HOLE that’s why it is only capable of → LYSING When the integrity is altered ions → Ions then are
RBCs but not nucleated cells able to pass freely out of the cell causing → lysis and
MAC begins hen C6 binds to → C5b there by loss of electrolyte and amino acid
stabilizing it followed by binding → C7 → C8 → C9 Destruction of target cells occurs through an →
INFLUX of water and a corresponding loss of
electrolytes and amino acid out of the cell
● MAC begins when the C6 binds to C5B there
stabilize followed by C789

The Classical complement cascade

It has 3 units:
➔ recognition
➔ activation
➔ Formation of MAC

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The similarity of the three pathway is they are all the

C4a, C3a, C5a → acts as ANAPHYLATOXIN same ng end point is the formation of → MAC or
membrane attack complex
C5a → it also acts CHEMOTAXINS
ALTERNATIVE PATHWAY
C2b → KININ like activity
Difference in ALTERNATIVE PATHWAY → it BYPASSES
C3b → used up in pathway, act as OPSONIN and use → C1,C2,C4
to FACILITATE PHAGOCYTOSIS
also called as BYPASS PATHWAY & PROPERDIN
PATHWAY
STARTS in → C3

● This represents the oldest of the C3 activating

pathways
● The pathway was originally named for the
protein → PROPERDIN (The pathway was
originally named for the protein properdin it
was though that properdin is the initiate
factor of the alternative)
●
● The serum proteins that are UNIQUE to this
pathway include → factor B and factor D.
● C1, C2, and C4, found in the classical pathway,
are not used at all in this system.
● C3 → is a key component of both pathways

● TRIGGERING substances for the alternative

pathway include → bacterial cell walls,
especially those containing
lipopolysaccharide; fungal cell walls; yeast;
viruses; virally infected cells; tumor cell lines;
and some parasites, especially
trypanosomes.
●
● All of these can serve as sites for binding the
→ complex C3bBb, one of the end products of
this pathway.

● Alternative pathway begins with the

→ activation of C3 and requires Factor B and
Factor D and Mg++ cation. → present in
NORMAL SERUM
The conversion in C3 → is the FIRST STEP in
alternative pathway
● C3 convertase- C3bbb
● C5 convertase- C3bbb3b

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when BACTERIAL POLYSACCHARIDE binds to the

circulating → C3b

Native C3 is circulating in SERUM normally, but the

initiating factor of alternative pathway is the binding
or PRESENCE of a BACTERIAL POLYSACCHARIDE

Native C3 is circulating in SERUM normally, but the

initiating factor of alternative pathway is the binding
or PRESENCE of a BACTERIAL POLYSACCHARIDE →
when there is presence of LIPOPOLYSACCHARIDE →
the circulating C3b binds to it.

C3 is spontaneously forming → C3b in serum

(because C3 is NOT stable that’s why usually
HYDROLYZE by WATER spontaneously activating a
small number of these molecules.→ C3b

C3b usually circulating in SERUM → C3 usually

hydrolyze by WATER to produce → C3b which then
bind to → FACTOR B (when there is presence of
LIPOPOLYSACCHARIDE)

presence of LIPOPOLYSACCHARIDE will trigger

attachment of C3b on cell membrane or bacterial cell
membrane and also binds to FACTOR B → and
together they attaches to the TARGET CELL SURFACE

Once C3b is bound to → FACTOR B it will SIGNAL

→ FACTOR D to cleave the → FACTOR B into
→ Ba & Bb → Ba will be cleave (it will remove) and →
Bb will remain = → C3bBb known as → C3 convertase

C3 convertase (C3bBb) STABILIZE by → PROPERDIN

PROPERDIN - is NOT the one initiate the but it is used

to STABILIZE the KEY ENZYME which is the
→ C3 convertase (C3bBb)

C3 convertase (C3) it will cleave → C3a & C3b

= producing more C3b

C3 cleaved into → C3a (will enter the circulation and

acts as an ANAPHYLATOXIN) & C3b will join the
Native C3 is NOT stable in → plasma.
complex → C3bBb3b known as → C5 convertase
Water is able to hydrolyze → C3 → spontaneously
activating a small number of these molecules.→ C3b C5 convertase (C5) it will cleave → C5a (will enter the
circulation and acts as an ANAPHYLATOXIN) & C5b
C3b spontaneously formed through the HYDROLYSIS of
WATER → C3b attaches alternative pathway is initiated C5b will be attach onto the LIPOPOLYSACCHARIDE
cell membrane of the antigen → and attract C6789
forming now → MEMBRANE ATTACK COMPLEX which
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is similar to the classical pathway → will punch holes IDENTICAL to classical

on the surface of the cell membrane causing → LYSIS DIFFERENCE → RECOGNITION UNIT
of the antigen (and then identical na after cleavage of
LECTIN RECOGNITION UNIT → MBL attaching to the →
C5) → the alternative pathway is now IDENTICAL to
MANNOSE OF BACTERIA → MBL interact to →
the classical pathway)
(MASP-2) SERINE PROTEASES → will be the one to
● LIPOPOLYSACCHARIDE → initiating factor in cleave → C4 & C2
alternative pathway
the rest is IDENTICAL to classical
●
LECTIN PATHWAY
● The MBL, MASP-1 and MASP-2 complex
analogous to Ab-C1qrs complex and leads to
This pathway is initiated by three proteins:
antibody-independent activation of C4,C2 and
➔ mannan-binding lectin (MBL), also known as C3.
mannan-binding protein (MBP) and
➔ two (2) mannan-bindng-lectin-associated
serine proteases (MASP-1 and MASP-2) → all ● The structure of MBL is similar to that of →
present in normal serum. C1q, and it is associated with three
MBL-serine proteases (MASPs): ○
INITIATING FACTOR when → MBL binds to→ certain
MASP-1,MASP-2, and MASP-3.
mannose residues on many bacteria and subsequently
interacts with MASP-1 and MASP-2.
● Once MBL binds to a cellular surface, MASP-2,
which is homologous to C1s, autoactivates.
ACUTE PHASE REACTANT (MBL) → which are
PROTEINS that binds → MANNOSE on the surface of
MANY BACTERIA/FUNGI → Subsequently binding of ● MASP-2 thus takes the active role in cleaving
MBL to the → MANNOSE OF BACTERIA → interacts C4 and C2, while the functions of MASP-1 and
with MASP-1 and MASP-2. MASP-3 are unclear at this time.

● MBL is → HOMOLOGOUS to → Ab-Ciqrs
complex which leads to the → activation of
C4, C2 and C3 in classical pathway
MBL → serve as Antibody
Mannose → Antigen
once naka bind si → MBL on MANNOSE → MASP-1
and MASP-2 is HOMOLOGOUS to → Ab-Ciqrs complex
which leads to the → activation of C4 & C2
MASP-2 - ang mag activate → C4 & C2 and
subsequently → C3 then IDENTICAL na siya sa classical
pathway
DIFFERENCE → walang C1 components and meron is
→ MBL, MASP-1 and MASP-2
● Once C4 and C2 are cleaved, the rest of the
pathway is identical to the classical pathway
● This the lectin pathway provides a means of
non specific protection against certain
pathogens before any antibody response can
be mounted.
ANTIBODY INDEPENDENT ACTIVATION
● C1qrs can also bind number of agents
including some retroviruses, mycoplasma,
poly-inosinic acid and aggregated IgG, and
initiate the classical pathway
C3a,C4a,C5a – anaphylatoxins C5a
MAC → C5b6789

similar ang LECTIN & CLASSICAL PATHWAY starting

→ C4 & C2
when C4 & C2 are cleave the rest of the pathway is
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C3bBb3bP.
→ In the classical and lectin pathways, C5 convertase
is made up of C4b2a3b. After C5 is cleaved, the
pathway is common to all

Convergence of the Classical, Alternative, and Lectin

pathways

Proteins of complement system

Classical Lectin Alternativ Lytic

pathway pathway e pathway pathway

Activation Mannan C3, Factors C5, C6, C7,

proteins: binding B and B, C8, C9
Convergence of the classical, alternative, and lectin C1qrs, C2, protein Properdin
pathways. The binding of C1qrs to two antibody C3, C (MBP),
molecules activates Mannan
➔ the classical pathway, whereas the alternative associated
pathway is started by hydrolysis of C3. serine
The lectin pathway is triggered by binding of MBP to protease
mannose on bacterial cell walls. (MASP,
MASP2)
→ MASP-1, MASP-2, and MASP-3 bind to form an
activated C1-like complex.
→ MASP-2 cleaves C2 and C4 and proceeds like the
Control proteins: C1-INH, C4-BP Factors I* and H,
classical pathway. Factor B and Factor D operate in the
DAF, CR1, etc. Protein S (vitronectin)
alternative pathway.
→ Although C3 convertase is formed differently in
each pathway, C3 is a key component in each one. The
C5 convertase in the alternative pathway consists of
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Biologically active products of complement

Plasma Complement Regulators activation

● Kinin production – C2b generated during the

classical pathway of C activation is a prokinin
which becomes biologically active following
enzymatic alteration by plasmin and causes
vascular permeability and edema.

● Excess C2b production is prevented by limiting

C2 activation by C1 inhibitor (C1-INH) also
known as serpin that dismantles the activated
C1qrs complex.

Deficiencies of Complement Components ● A genetic deficiency of C1-INH results in an

overproduction of C2b and is the cause of
hereditary angioneurotic edema

Anaphylatoxins

● C3a and C5a are all anaphylatoxins that cause

basophil/mast cells degranulation and smooth
muscle contraction.

● An uncontrolled production of these

anaphylatoxins can lead to pathological
consequences.

● These anaphylatoxins are normally inactivated

by carboxypeptidase N (C3a-INA).

Chemotactic factor

● C5a and MAC (C5b67) are both chemotactic.

● C5a is also a potent activator of neutrophils,

and macrophages and thus amplifies
nonspecific immunity.

● It also causes induction of adhesion molecules

on vascular endothelial cells and hence
promotes diapedesis.

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 IMMUNOLOGY AND SEROLOGY
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1st SEM, 2021

Initiates attraction of MACROPHAGES, NEUTROPHILS,

MONOCYTES, DENDRITIC CELLS to the antigen.
➔ The binding of C5b to monocyte will cause
them to undergo oxidative BURST causing
incomplete. Biological properties of C-activation products and
➔ Production of HYDROLYTIC enzymes as well as their regulatory molecules
CHEMOTACTIC FACTOR
C3a & C5a are rapidly inactivated by enzymes Component Biologic Effect Controls
carboxypeptidase to localize and control regulate their
activity
effects.
C2b (prokinin) Accumulation Edema C1-INH
Opsonins of body fluid

C3a Basophil and Anaphylaxis Carboxypeptid

● C3b and C4b on the surface of microorganisms (anaphylatoxin) mast cell ase-B
attach to C-receptor (CR1) on phagocytic cells degranulation; (C3a-INA)
enhanced
and promotes phagocytosis. vascular
permeability;
smooth
● This binding facilitates phagocytosis and muscle
clearance of foreign substances or cellular contraction
debris, which is one of the key functions of
Induction of Immuno
the complement system. suppressor regulation
T-cells
3 phase for each of the main pathway
C3b and its Opsonization; Phagocytosis Factors H and I
products phagocyte
1. Initiation/ Recognition activation
➔ staring point, recognition, initiates the
C4a Basophil and Anaphylaxis C3a-INA
cascades (anaphylatoxins mast cell
degranulation;
2. Amplification/ Activation enhanced
vascular
➔ activation of other complement components permeability;
smooth
3. MAC muscle
contraction
➔ cell lysi
C4b Opsonization Phagocytosis C4-Bp, Factor I

C5a Basophil and Anaphylaxis C3a-INA

(anaphylatoxin mast cell
; chemotactic degranulation;
factor) enhanced
vascular
permeability;
smooth
muscle
contraction

Chemotaxis; Inflammation immunoregula

neutrophil delayed tion
aggregation; anaphylaxis
oxidative
metabolism
stimulation.
Stimulation of
leukotriene
release;
induction of

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 IMMUNOLOGY AND SEROLOGY
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1st SEM, 2021

helper Tcells

C5b67 Chemotaxis; Inflammatory C3a-INA

attachment to lysis of by
other cell stander cells
membrane

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 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

HYPERSENSITIVITY
Hypersensitivity reaction or Allergic reaction are all
AT THE END OF THIS UNIT, THE STUDENT
cause by IMMUNE MECHANISM (sa kagustuhan ng
WILL BE ABLE TO:
immune system na ma neutralize ang antigen pati
● Differentiate the immunologic mechanism
surrounding tissues, normal antigen, and organs
involved in the different types of
nadadamay nagkakaroon sign and symptoms becos of
hypersensitivity reactions in terms of antibody
your immune responsiveness called →ALLERGIES or
involvement, complement involvement,
ALLERGIC REACTION
antigen triggers, and timing of the response.
● Correctly assess and associate the type of Hypersensitivity reactions can be classified into
hypersensitivity with regard to disease or 4 difference types
condition.
4 common types of hypersensitive:
OVERVIEW
➔ Type 1
● Hypersensitivity refers to UNDESIRABLE
➔ Type 2
REACTIONS produced by the → normal
➔ Type 3
immune system.
➔ Type 4
● Hypersensitivity reactions require a
pre-sensitized (immune) state of the host. Type I Hypersensitivity
● Hypersensitivity reactions can be divided into
four types: type I, type II, type III, type IV,
based on the mechanisms involved and time Type I Hypersensitivity also known as → IMMEDIATE
taken for the reaction. or ANAPHYLACTIC TYPE OF HYPERSENSITIVITY
REACTIONS
● type I, type II, type III, type IV → These
Type I Hypersensitivity commonly thought as →the
hypersensitivity reactions have different
ALLERGIES
mechanisms involved and the time taken for
the reaction to occur. Usually the reaction that is involved in Type I
Hypersensitivity or the reaction that result from these
● Hypersensitivity reaction → reactions that are immune responsiveness from type 1 primarily IgE
DAMAGING, causing discomfort in the body antibody, involves →SKIN that’s why yung symptoms
and sometimes can become FATAL and these
nito causes → URTICARIAL or ECZEMA also may
are IMMUNE MECHANISM of a normal
involve your EYES causing →CONJUNCTIVITIS or
immune system , → but sometimes your
sometimes allergies can be exhibited by pamamaga ng
immune system or the antigens in the body
persistent too long that’s why the immune mata or redness of eye and itchy eyes or conjunctivitis
mechanism also DAMAGES the surrounding as well as NASOPHARYNX that why some allergies can
tissues and organ that causing damaging and trigger RHINITIS or RHINORRHEA
discomfort. The reaction may involve:
HYPERSENSITIVITY SKIN → (urticarial and eczema)
● Heightened state of immune responsiveness
EYES → (conjunctivitis)
Allergic reactions → eto din ang hypersensitivity
reactions. NASOPHARYNX→ (rhinorrhea, rhinitis)

● It is an EXAGGERATED RESPONSE to an BRONCHOPULMONARY TISSUE →(asthma)

innocous antigen that results in → gross
tissue changes that are DELETERIOUS, GASTROINTESTINAL TRACT →(gastroenteritis)
DAMAGES or can be FATAL to the host.

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 IMMUNOLOGY AND SEROLOGY
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● An immediate hypersensitivity, SUDDEN tendency to produce a HUGE amount of IgE and

ALLERGIC response mediated by antibodies,
primarily → IgE
●
● Antigens that trigger formation of IgE are
called → atopic antigens, or allergens.
● ATOPOS which means “OUT OF PLACE” wherein it
● Atopy → refers to an inherited tendency to
respond to naturally occurring inhaled and
ingested allergens with continued production
of IgE (kaya usually yung mga may ASTHMA
sila yung may mga sensitive skin, allergies sa
iba’t ibang mga bagay becos sakit ito and that
is known as → ATOPY usually this a
HEREDITARY
●
Type I Hypersensitivity - very sensitive or kahit
konting antigens lang yung ma-inhale or ingested you
have an inherited tendency to produce an INCREASE
produce of IMMUNOGLOBULIN E (IgE) which causes
→ALLERGIC SIGNS & SYMPTOMS or TYPE 1
HYPERSENSITIVITY SIGNS & SYMPTOMS
Usually the reaction of type 1 hypersensitivity
sometimes pamumula lang ng balat or pamamantal
nagkakaroon ng sipon but can also result in MAJOR
SYMPTOMS such as →→ ASTHMA nagkakaroon ng
blockages sa airflow called →Bronchoconstriction and
sometimes nagkakaroon ng ANAPHYLACTIC which can
lead to → death
Kahit na small amount of concentration of antigen
yung makapasok sa person they have this inherited
tendency to to produce a →LARGE AMOUNT of
IgE.
IgE have receptors in MAST CELLS and BASOPHILS. This
specific IgE will bind to MAST CELL and BASOPHILS
becos IgE have receptors to mast cells and basophils.
Pag nag bind na sila upon secondary exposure the IgE
will CAUSE and bind the antigen causing
DEGRANULATION of mast cells and basophils and the
GRANULES that will released will cause the SIGN and
SYMPTOMS of type 1 which can involve the → SKIN,
PHARYNX, BRONCHUS and GIT
Antigen that trigger formation are called → ALLERGIES
and the ATOPY is the disease or the inherited
response to allergen.
Atopy → term which derived from the greek know as
refers to an inherited tendency to develop a classical
allergic response to → naturally occurring ingested
ALLERGENS
● It involves →SECONDARY EXPOSURE to an
offending allergen that bounds to→ mast
cell-fixed IgE
Key terms of Type I Hypersensitivity
➔ Allergies
➔ Immediate
➔ Anaphylactic type
➔ Primarily mediated by IgE which is fixed to
MAST CELLS & BASOPHILS that’s why type 1
hypersensitivity involves → CELL BOUND
ANTIBODIES
➔ Secondary exposure → to an ALLERGEN to
antigen is an allergen.
Secondary exposure sa pangalawang pangalawa beses
ma exposure dun sa allergen dun palang magkakaroon
ng → type 1 hypersensitivity
→ kasi yung PRIMARY EXPOSURE gagawa palang ng
IgE so after mag form ng plasma cell mag porpoduce
ng SPECIFIC IgE for that ALLERGENS, so after mag
create ng specific IgE for that offending antigen during
primary exposure, those specific IgE will look for mast
cell and basophil then mag mabind sila doon,
→ then upon SECONDARY EXPOSURE they will AGAIN
bind upon secondary exposure to that particular
antigen kung saan na expose primarily it will again
bind kasi SPECIFIC na siya for that ALLERGEN so ibibind
na yung allergen
→then dahil naka fixed yung FC portion (for the host
cell) niya sa MAST CELL yung FAB portion naka bind sa
allergen/antigen on secondary exposure and upon
binding IgE sa antigen - ma trigger si MAST CELL
kailangan labanan niya yung ANTIGEN mag

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DEGRANULATE yung mast cell releasing so when
antigen crosslink two
Criteria of secondary link needs to → cross-links TWO
adjacent IgE molecules
→ The antigen cross-links TWO adjacent IgE
molecules, leading to → DEGRANULATION of the mast
cells, with release of pre-formed VASOACTIVE
MEDIATOR such as:
● histamine
● ECF-A (Eosinophil chemotactic factor)
● neutrophil
● chemotactic factor
● tryptase as well as → newly synthesized na
mamafoform palang becos of the triggers will
cause all the symptoms of type 1
hypersensitivity reactions sa sobrang lakas ng
binigay nilang immune response kaya
nagkakaroon ng symptoms → mediators such
as prostaglandins and leukotrienes.
ALL OF THESE ONE CAUSES SYMPTOMS OF TYPE 1
HYPERSENSITIVITY → manifestation in seconds to
minute after secondary exposure kaya nga IMMEDIATE
HYPERSENSITIVITY REACTION ang type 1 becos within
SECONDS/ MINUTES mag DEGRANULATE maglalabas
ng HISTAMINE and other vasoactive mediator kaya
nagkaroon ng symptoms
VASOACTIVE MEDIATOR - lalaban sila or they will act
or toxic to the particular allergen kaya lang sa sobrang
EXAGGERATED ng immune response →
VASOACTIVE MEDIATORS CAN CAUSE THE
SYMPTOMS OF TYPE 1 HYPERSENSITIVITY:
➔ EDEMA
➔ can cause RETINA URTICARIA
➔ CAPILLARY PERMEABILITY
➔ MUCOUS GLAND SECRETION causing
ALLERGIC RHINITIS
➔ ACID production on stomach causing
GASTROENTERITIS
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62
➔ contraction of SMOOTH MUSCLE causing
BRONCHOCONSTRICTION
HISTAMINE - can increase the contraction of SMOOTH
MUSCLE that’s why masakit yung tummy at
naapektuhan yung GIT
→ also HISTAMINE can increase CAPILLARY
PERMEABILITY and can increase MUCOUS GLAND
SECRETION that’s why excess histamine on the body
can cause → ALLERGIC RHINITIS (sinisipon pag
nagkaroon ng allergic reaction sa isang allergen)
HISTAMINE - also cause RETINA URTICARIA
(namamantal/nangangati)
can also increase the ACID production in stomach
causing GASTROENTERITIS and can cause contraction
ng smooth muscle causing BRONCHOCONSTRICTION
● The antigen cross-links TWO adjacent IgE
molecules, leading to→ degranulation of the
mast cells, with release of pre-formed
vasoactive mediators such as: histamine,
ECF-A, neutrophil, chemotactic factor and
tryptase as well as newly synthesized
mediators such as prostaglandins and
leukotrienes.
IMMUNOLOGIC MECHANISM
IMMUNOLOGIC MECHANISM - It is activity of IgE
● The regulation of IgE production appears to
be a FUNCTION of a subset of T cells called
→ type 2 helper cells (Th2).
T-helper 1 - T cells activation
T-helper 2 - for B Cell activation
● In people with allergies, the immune response
is SHIFTED and Th2 cells → PREDOMINATE.
that’s why usually ang nag rereact during a
patients with allergy naka SHITED or naka
activate ang → Th2 response.
In a normal immune response to antigen there is an
appropriate balance between the activity of T-helper 1
and T-helper2 → which results in a protective
immunity that DO NOT HARM the HOST
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

→ but in people with allergies the immune response is ● IL-4 and IL-13 → activate transcription of the
SHITED and T-helper2 activity or T-helper 2 response EPSILON GENE in B cells when they bind to
predominant immune response that’s why the specific receptors
T-Helper 2 type of response - results in production of
SEVERAL CYTOKINES.
Cytokines → produce Interleukin 4,5,9,13 - these
cytokines are responsible for final differentiation that
occurs in B cells that is why the → T helper2 is
involved in B cells activation to become →
PLASMA CELLS.
These cytokines - initiate the transcription of the gene
that codes for epsilon

● This Th2 type of response results in

production of several cytokines, including
IL-4 and IL-13
● These CYTOKINES are responsible for the final
differentiation that occurs in B cells, initiating
the transcription of the GENE that codes for →
the epsilon-heavy chain of immunoglobulin
molecules belonging to the IgE class. That's
why patients with allergies tend to have
increase amount of IgE that why madami
napoproduce na IgE.
→ SENSITIZATION PHASE
SENSITIZATION PHASE → primary response and
activation is the → SECONDARY EXPOSURE kasi
kailangan muna siyang ma expose and that is known a
→ SENSITIZATION
for the body to create specific or antigen specific IgE
and upon SECONDARY EXPOSURE to that antigen ma
activate na siya again na that is
→ ACTIVATION PHASE

Anti histamine →serves as BLOCKERS for the receptor SENSITIZATION PHASE - stage wherein the IgE
of histamine so that hindi na siya makapag induce ng antibody attaches to high specific or high affinity
symptoms sa katawan. receptors which bind to the FC portion of IgE so mag
bibind/mag form ng IgE or antigen specific IgE that
IL-5 and IL-9 →are involved in the development of attaches to the MAST CELLS & BASOPHILS
EOSINOPHILS, that's why eosinophils are increased or
● Formation of antigen-specific IgE that attaches
have a → EOSINOPHILIA - increase allergic reactions
to mast cells.
IL-4 and IL9 →promote development of MAST CELLS.
● IgE antibody → attaches to high-affinity
● IL-4, IL-9, and IL-13 all → act to stimulate
receptors called FcεR (FC EPSILON RECEPTOR
overproduction of MUCUS that also
GENE), → which bind the fragment
contributes that’s why nagkakaroon ng
crystallizable (Fc) region of the epsilon-heavy
RHINITIS during allergic reaction, a
chain.
characteristic of most allergic reactions.
FC portion - bind complement, monocytes,
● This propensity to secrete cytokines that
macrophages, and that is also the binding site of
promote production of IgE is LINKED → to a
MAST CELLS & BASOPHILS
gene locus on chromosome 5 that encodes
cytokines IL-3, IL-4, IL-5, IL-9, IL-13, and
FC portion of IgE - meron silang receptor to mast cells
granulocyte-monocyte colony stimulating
that known as →FC EPSILON RECEPTOR doon mag
factor (GMCSF). All of these cytokines are KEY
mag bind si IgE yung kanilang FC REGION
to a switch to a → Th2 response.

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● Once bound, IgE serves as → an antigen
receptor on mast cells and basophils to bind
the antigen, so indirectly kaya na ngayon mag
bind ng mast cell and basophils kasi meron sila
IgE though FAB portion of the IgE mast cell
and basophil can now bind an antigen
● Subsequent binding of ALLERGEN to →IgE -
sensitized mast cells and basophils triggers →
degranulation with release of inflammatory
mediators
Fc portion - portion of IgE na mag bind sa FcεR (FC
EPSILON RECEPTOR GENE) at nakikita sa basophil and
mast cell
→ binding of IgE to basophil and mast cell will increase
the half-life of IgE 2 to 3 days lang normally pag
UNBOUND
→ pag naka BOUND siya sa basophil and mast cell, it
will increase the half life of IgE up to at least 10 days
MAST CELL - are the PRINCIPAL effector cells of
immediate hypersensitivity and these cells found →→
throughout the body and most organs but they to
concentrate on BLOOD VESSELS, LYMPHATIC,
GRANULAR TISSUE → they have cytoplasmic granules
which serves as inflammatory mediator which are
PRE-FORMED (pre-formed means gawa na kahit
walang exposure meron ng histamine or other
pre-formed inflammatory mediator inside the mast
cell
BASOPHILS - they differ in terms of their appearance
and functions. Present on PERIPHERAL BLOOD &
BLOOD VESSEL and they only 1% of the total WBC
pool; they have FEWER but have LARGER GRANULES
compared to mast cells.
but BOTH of which the BASOPHILS & MAST CELLS →
release PRE-FORMED inflammatory/vasoactive
mediator which causes the → TYPE 1
HYPERSENSITIVITY OF SIGNS AND SYMPTOMS
ACTIVATION PHASE
→after ng primary exposure so andoon na
ALLERGEN present in → T-helper 2 involve in B cells
activation that’s why B cells will produce or will
become → activated and become PLASMA CELLS →
PLASMA CELLS now dahil nga nagkakaroon ng SHIFT
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64
into 2 T-helper 2 response yung cytokines involved
such as 13 & 44 in production of different symptoms
of hypersensitivity and as well as synthesis of an
EPSILON GENE → nagkakaroon ng overproduction of
our IgE→ SENSITIZATION PHASE
ACTIVATION PHASE →begins when there is a
subsequent or secondary exposure to the SAME
allergen and the ALLERGEN cross link the cell bound
IgE → needs 2 adjacent cell bound IgE mag bibind
yung allergen which are now trigger the release of the
→PHARMACOLOGICALLY ACTIVE SUBSTANCES or also
known as VASOACTIVE SUBSTANCES which are found
on MAST CELLS & BASOPHILS
● Adjacent cell-bound IgE molecules are
cross-linked by a bivalent or multivalent
antigen, causing aggregation of the surface
FcεRI receptors.
mag DEGRANULATE lang siya if the allergen →
CROSS-LINKED the FC IgE receptor →kailangan
CROSS-LINKED para ma trigger ang DEGRANULATION
releasing the→ VASOACTIVE MEDIATORS
once nag CROSS-LINKED IgE receptor na naka bound
sa MAST CELLS it will initiate a series complex
intracellular signaling events involving →
PHOSPHORYLATION in INFLUX OF CALCIUM during the
degranulation process
● Mast cell degranulation is preceded by
increased → Ca++ influx triggers rapid
degranulation of mast cell and basophils
which release the chemical mediator that
have been previously made and stored in the
granules this is known as →PRIMARY
MEDIATOR
HISTAMINE - most abundant preformed/primary
mediator and causes the SYMPTOMS
● Release of primary mediators
PRIMARY MEDIATORS:
➔ Heparin
➔ ECF-A (Eosinophil chemotactic factor)
➔ chemotactic factor
➔ tryptase
 IMMUNOLOGY AND SEROLOGY
Introduction (WEEK 2/ LEC)
1st SEM, 2021

➔ other proteinase that are toxic to these

antigen

within 30 to 60mins after exposure meron na agad

SYMPTOMS and that is becos of the →
PRIMARY MEDIATOR

LATE PHASE ACTIVATION

narerelease lang to pag naexpose na sa ANTIGEN

● Synthesis of NEWLY FORMED, or SECONDARY

MEDIATOR → include:
➔ platelet-activating factor (PAF);
➔ prostaglandin (PG) D2;
➔ leukotrienes (LT) B4, C4, D4, and E4;
➔ cytokines
These mediator are responsible for the LATE PHASE
ALLERGIC REACTION that can be seen on some
individual from → HOURS to 8hrs after exposure to
the antigen.

● Eosinophils → play an important role in the

late-phase reaction so during an allergic
reaction there are CYTOKINES such as → IL-5
- that are released from Thelper stimulation
that’s why increase din ang EOSINOPHILS or →
EOSINOPHILIA which is sign that you have an
ongoing allergic reaction becos of the release
of the → pre-formed/newly synthesized
mediator
●
● The number of FcεRI receptors on eosinophils
INCREASES during the allergic response, and
the eosinophil is stimulated to release a
variety of toxic molecules and inflammatory
mediators from its granules.

Pharmacologic mediators of immediate

hypersensitivity
Clinical Manifestations: Type 1

● SYMPTOMS depend on such variables as route

of antigen exposure, dose of allergen, and
frequency of exposure.
● Rhinitis is the most common form of →
ATOPHY, or allergy
○ Symptoms include paroxysmal
sneezing; rhinorrhea, or runny nose;
nasal congestion; and itching of the
nose and eyes

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 IMMUNOLOGY AND SEROLOGY
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1st SEM, 2021

→Hay fever, Asthma, Food allergies, Dermatitis ➔ Animal dander

(Urticaria or eczema), Angioedema usually AIRBORNE

Hay fever - cause by spring temperate climate Clinical Manifestations: Type 1

Asthma - respiratory disorder wherein ang

mechanisms involve type 1 hypersensitivity, na inhale
mo yung particles and it reaches the→ lower
respiratory tract can lead to→
BRONCHOCONSTRICTION

Asthma - means PANTING/BREATHLESSNESS

Food allergies - symptoms are NOT usually in skin, the

symptoms are stomach ache, vomiting and diarrhea.

Dermatitis - local inflammation of the SKIN usually the

reaction manifest either →URTICARIA or ECZEMA

Urticaria or Hives - (pangangati, pamumula, at

pamamantal) it appears within a MINUTES after
exposure to an allergen and characterized by SEVERE
ITCHING, REDNESS cause by VASODILATION and
LEAKAGE OF FLUID surrounding the area/REDNESS
around the center of the lesion.
→ usually that is known as WHEAL & FLARE REACTION
kaya pag nag SKINTEST tayo if may allergies ka doon sa
ininject magkakaroon ng WHEAL & FLARE REACTION -
redness around the center

Angioedema - reaction occurred DEEPER on the

dermal tissue

Rhinitis - cause by VASOACTIVE MEDIATORS Clinical Manifestations: Type 1

Can cause a SEVERE SYSTEMIC RESPONSE also known The most SERIOUS discomfort or type of Clinical
as →ANAPHYLACTIC/ANAPHYLAXIS SHOCK which can Manifestations of type 1 is → ANAPHYLAXIS may
lead ASPHYXIA (blockage of airway) leading to FATAL if not treated properly
FATALITY & DEATH
● Anaphylaxis → is the most severe type of
SYSTEMIC → means very fatal & deadly allergic response because it is an acute
SKIN TEST - tinitignan dito if may allergies ka on a reaction that simultaneously involves multiple
certain drug organs →usually pag nagkaroon na ng
BREAKDOWN ORGANS called as →
Allergens that causes Rhinitis: ANAPHYLACTIC SHOCK or MULTIPLE ORGAN
➔ Pollen FAILURE wherein hindi na nag function yung
➔ Mold spores mga organs and it is FATAL.
➔ Dust
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ANAPHYLAXIS - is coined and studied about the
immediate hypersensitivity by → Charles Richet &
Paul Portier
● Typical agents that induce anaphylaxis include
venom from bees, wasps, and hornets; drugs
such as penicillin; and foods such as shellfish,
peanuts, and dairy products
Clinical signs of anaphylaxis begin within →MINUTES
after antigenic challenge
ANAPHYLACTIC SHOCK - VERY SERIOUS & FATAL
ANAPHYLACTIC may result as → ASPHYXIATION
(Asphyxiation occurs when the body does not get
enough oxygen) parang nalulunod becos of UPPER
AIRWAY EDEMA kasi HISTAMINE and other vasoactive
mediator causing →BLOCKAGE/AIRFLOW
OBSTRUCTION becos of smooth muscle contraction
cause by HISTAMINE
● Signs may include bronchospasm and
laryngeal edema, vascular congestion, skin
manifestations such as urticaria (hives) and
angioedema, diarrhea or vomiting, and
intractable shock because of the effect on
blood vessels and smooth muscle of the
circulatory system.
Type I Hypersensitivity → can also cause a LATEX
SENSITIVITY as well as Type 4
Type 4 - contact DERMATITIS
Type 1 & type 4 - both cause reactions of antigen in
natural rubber latex which are found in GLOVES
Type 1 & type 4 - latex sensitivity
TREATMENT
● AVOIDANCE of known allergens is the → first
line of defense
● Localized allergic reactions, such as hay fever,
hives, or rhinitis, can be treated with →
ANTIHISTAMINES which block histamine
receptors and decongestants
● Systemic anaphylaxis is a medical emergency
that requires timely → injection of
EPINEPHRINE
DRUGS FOR ASTHMA:
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67
➔ BRONCHODILATOR
➔ SALBUTAMOL
➔ THEOPHYLLINE
➔ ANTI HISTAMINES
SEVERE ASTHMA DRUG:
➔ LEUKOTRIENE
➔ RECEPTOR ANTAGONIST
➔ BLOCKERS OF LEUKOTRIENE (para di mag
cause ng symptoms)
➔ MUSCLES STABILIZER
➔ STEROID - to block the inflammatory cells
➔ EPINEPHRINE - powerful vasoconstrictor
● Chromolyn sodium inhibits mast cells
degranulation, probably by inhibiting Ca++
influx.
Hyposensitivity or Allergy immunotherapy (AIT)
● Another treatment modality which is
successful in an number of allergens,
particularly to insect, venoms and to some
extent pollens.
The goal of Allergy immunotherapy → to induce
immune tolerance (administer the specific allergen for
which the patient is allergic) administer of gradually
INCREASING OF DOSES of the allergen overtime
meaning sinasanay mo siya and called →
HYPOSENSITIVITY (small amount of allergen in
increasing dosage so that unti-unti mablock yung IgE)
● This therapy is believed to SHIFT the patient’s
immune response to the allergen to a →
Th1-type of response and to induce the
development of T regulatory cells (Tregs) that
release → IL-10
IL-10 - is a CYTOKINES that REDIRECT immune system
to produce allergens specific IgG - for blocking
antibodies
IgG4 - para dI magkaroon ng DEGRANULATION and
tends to BLOCK the attachment of the antigen to the→
IgE
● The mechanism is not clear but there is a
correlation between appearance of IgG
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(blocking) antibodies and relief from
symptoms.
● Suppressor T cells that specifically inhibits IgE
antibodies may play a role.
The standard practice of Allergy immunotherapy is to
administer the allergens subcutaneously under the →
SKIN over 3 to 5 years
DIAGNOSIS TEST
● For immediate hypersensitivity include skin
(prick/ cutaneous testing and intradermal)
tests, measurement of total IgE and specific
IgE antibodies against suspected allergens.
In vivo test → cutaneous testing and intradermal - can
detect hypersensitivity to WIDE VARIETY of the
inhaled food allergens
Prick/ cutaneous testing → the doctor uses a needle
and picking devices to introduce a small drop of the
allergen on the upper layer of the skin (usually ito
yung SKIN TEST - to know if we have allergies to the
specific allergens)
POSITIVE RESULT →it’s a WILL AND FLARE REACTION
wherein pag nagkaroon ng redness on the circle dun
sa site where you have a inoculated allergens means
it’s a POSITIVE sign, nagkaroon ka ng →
URTICARIA/redness (degranulate happen)
Intradermal test - uses a GREATER amount of antigen
and it’s MORE sensitive than prick or cutaneous
testing. Usually performed if the prick test are
NEGATIVE to confirm. WILL AND FLARE REACTION →
positive result to determine the score of the both test.
In vitro test →RIST & RUST - which is the
measurement of total IgE and specific IgE antibodies
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● Total IgE and specific antibodies are
measured by a modification of enzymes →
immunoassay (ELISA).
Measurement of the TOTAL/AMOUNT of IgE and
specific IgE antibodies the test we used is known as →
RIST OR THE RADIO IMMUNO SORBENT
To detect the SPECIFIC IgE which is antibody IgE that’s
specific to particular antigen antibody →RAST
(RADIOALLERGOSORBENT TEST)
● Increased IgE levels are indicative of →ATOPIC
CONDITION.
RIST OR THE RADIO IMMUNO SORBENT - defect the
TOTAL of IgE uses an specific IgE/ uses a SOLID PHASE
ANTI IgE (nakadikit, siya yung pang catch)
Immunoassay (ELISA) - we use SOLID PHASE ANTIGEN
Type II Hypersensitivity
→ also known as antibody mediated CYTOTOXIC
hypersensitivity reaction
SYMPTOMS →CYTOLYSIS - sinisira yung cells
nagkakaroon ng CELL LYSIS
● Reactions that produce cell damage which is
mediated by complement fixing antibodies
(IgG & IgM) → directed against cell surface
antigens.
Type 1: ALLERGEN
Type 2: CELL SURFACE ANTIGEN - antigens that found
on cell surface, example is →antigen on the RED CELL
(ANTIGEN A & ANTIGEN B as well as D antigens)
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● The underlying mechanism involves IgG and

IgM antibodies directed against antigens
found on cell surfaces.

● Clinical examples that involve destruction of

cells by type II hypersensitivity include blood
transfusion reactions, → hemolytic disease of
the newborn, autoimmune hemolytic anemia,
Drug-induced hemolytic anemia,
granulocytopenia and thrombocytopenia

SYMPTOMS →Hemolytic transfusion reactions and

other blood hemolytic reactions

The reaction time is minutes to hours. Reaction is →

MINUTES to HOURS transfusion reaction
Cell lysis/Cell damage can occur by several different
mechanisms, some of which involve
complement/activation of complement promote
opsonization & phagocytosis as well as antibodies:
CLINICAL MANIFESTATIONS: Type II
● A second possible effect of type II
hypersensitivity is that the cell surface
antibody can inhibit the function of a cell

1. Activation of the classical pathway of ○ An example of this effect occurs in the

complement can lead to the formation of the autoimmune disease → myasthenia
membrane attack complex and cell lysis. gravis - example of type 2
nagkakaroon ng inhibition of
2. Coating of the cell surface by antibodies can NEUROMUSCULAR function during →
promote opsonization and subsequent myasthenia gravis
phagocytosis of the cells

3. Cell damage can result from the → mechanism ● Transfusion Reactions

of antibody dependent cellular cytotoxicity
(ADCC) → wherein ang mag bibind sa FC
portion are MACROPHAGE & NATURAL KILLER
CELLS releasing CYTOTOXIC enzymes that will
destroy cells.
○ The ABO blood group is of primary
importance in considering
transfusions
○ If a patient is given blood for which
antibodies are already present (wrong
match), a transfusion reaction occurs.

○ Delayed hemolytic reactions (IgG)

involved → RH, Kell, Duffy, Kidd occur
within the first 2 weeks following a
transfusion and are caused by an
anamnestic response to the antigen
to which the patient has previously
been exposed

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 IMMUNOLOGY AND SEROLOGY
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ABO, RH, Kell, Duffy, Kidd system → causes/ involve in
HEMOLYTIC TRANSFUSION REACTION
ABO HEMOLYTIC TRANSFUSION REACTION - IgM & IgG
but majority antibodies form during ABO blood group
hemolytic transfusion reaction is → IgM
can cause also→ disseminated intravascular
coagulation
SYMPTOMS:
● Fever
● Chills
● Back Pain
● Tachycardia - refers to heart rate that too fast
● Hemoglobinuria
Hemolytic Disease of the Newborn
antibody involved →IgG or what is known as Anti-D
which are antibodies to D antigen/RH
Hemolytic Disease of the Newborn involved
RH negative mother and RH positive child
● This appears in infants whose mothers have
been exposed to blood group antigens on the
baby’s cells that differ from their own.
● Severe HDN is called → erythroblastosis
fetalis
● kilala ang D antigen dahil siya ay RH negative →
● D antibody is usually involved
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● Other antibodies associated with HDN include
anti-c, anti-C, anti-E, anti-e, and less
commonly those associated with the Kell,
Duffy, and Kidd blood groups
ABO → cause is MILDER
MECHANISM OF HEMOLYTIC DISEASE OF NEWBORN:
usually the FIRST CHILD is UNAFFECTED - dun palang
gumawa ng ANTI-D
SECOND CHILD (RH positive) - dahil gawa na ang
ANTI-D mag cross sa placenta at sisirain yung red cells
nung baby
→ During the first pregnancy if the MOTHER is RH
negative and the BABY is RH positive
RH positive - meron D antigen sa red blood cells
RH negative - walang D antigen
so during FIRST PREGNANCY meron tinatawag na
→ fetal maternal hemorrhage
wherein yung rbcs from the baby ENTER the
circulation of the mother →pwede during GESTATION
→ or during the birth/delivery → the later part of
gestation meron nangyayari → fetal maternal
hemorrhage wherein the blood cells the baby reaches
the SYSTEMIC or the circulation of the mother
→dahil ang red cell ng baby ay D antigen sa surfaces
then si mother ay wala so FOREIGN sa kanya yung D
antigen →ang gagawin ng immune
mechanism/immune system ni mother → dahil di niya
ipresent yun sa mga Naive B cells will become Plasma
cells and secrete ANTI-D or antibody against the D
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antigen kung saan na expose during the first

pregnancy
→ so ngayon si mother meron na siyang ANTI D na
produce kasi na EXPOSE siya during fetal maternal
hemorrhage → mainly the ANTI D antibody are of
IgM type → kasi primary exposure first born naive B
cell ang na produce that’s why yung IgM → will not
cross the placenta kaya NO HARM dun sa ating first
baby
→ After that some of the ANTI D that was produce will
become memory cells or some of the plasma cell will
become memory cells that will now produce the →
IgG type of this ANTI D
Now sa → SECOND baby positive padin that’s why the
red blood cells nung baby ay may D antigen ngayon
nag form → ng memory B cells that can produce ANTI
D that are IgG now → IgG can cross the placenta,
during pregnancy may fetal maternal hemorrhage
AGAIN → so mapupunta ngayon ulit yung rbcs upon
subsequent exposure may activate si B cells at
memory cell na ag mag activate kasi active immune
system or meron ka ng NATURAL ACTIVE ADAPTIVE
IMMUNITY wherein na expose kana previously kaya
nakaproduce kana ng memory B cells that why
increase ang magnitude of response at mas mabilis
mo ng ma attach ang mga antigens
REMEDY: Usually kapag RH NEGATIVE ang mother and
nag deliver siya ng RH POSITIVE baby binibigyan na
siya ng → RhoGAM or ANTI D antibodies during
delivery usually binibigay na ang RhoGAM 72 hours →
after delivery para yung mga red blood cells during
fetal maternal hemorrhage ay ma bind na ni RhoGAM
at hindi ma activate ang → B cells at para di na
gumawa ng ANTI D so that during SECOND
PREGNANCY wala ng memory B cells that can cross
the placenta and cause → hemolytic disease of the
newborn
OTHER CLINICAL MANIFESTATIONS: Type II
● Autoimmune Hemolytic Anemia
Symptoms include
➔ malaise
➔ lightheadedness
➔ weakness
➔ unexplained fever
➔ pallor
➔ possibly mild jaundice
● Drug induced → Hemolytic Anemia
● Idiopathic autoimmune hemolytic anemia

→dahil fetal maternal hemorrhage na activate ulit

ang ating memory B cells secrete na ngayon yung
memory cells na ANTI D at sisirain niya lahat and then
IgG can cross the placenta →that's why sisirain niya
lahat ng D ANTIGEN kasi nga siya ay ANTI D specific for
the D ANTIGEN → so sisirain niya ang D ANTIGEN Testing for Type II Hypersensitivity
which i found with red cell of the baby that’s why
lahat ng red cells niya masisira icocoat and that's TYPE A. Direct antiglobulin
2 HYPERSENSITIVITY REACTION → the baby will have
ANEMIA it depends sa severity the baby cannot ● Testing (DAT) is performed to detect
CONJUGATE BILIRUBIN yung baby wala pa siya mga transfusion reactions, hemolytic disease of
enzymes sa liver niya or poorly developed pa that’s the newborn, and autoimmune hemolytic
why hindi niya ma conjugate yung bilirubin anemia.
In vivo: we use DAT - detect transfusion reactions;
→that’s why BILIRUBIN nagkakaroon ng excess INVESTIGATE if may nangyaring sensitization inside the
causing JAUNDICE OF THE FETUS and bilirubin body
deposits on the EYES, BRAIN causing →KERNICTERUS -
symptoms of hemolytic disease of the newborn and if
severe na can lead to →death of the baby
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In vitro: IAT - prior to blood transfusion and is used to
detect red cell antibodies in patient serum; cross
matching in VITRO
● Polyspecific antihuman globulin is a mixture of
antibodies to IgG and complement
components such as C3b and C3d, and it is
used for initial testing.
● If the test is positive, then it should be
repeated using monospecific antiIgG,
anti-C3b, and anti-C3d to determine which of
these is present.
● If an autoimmune hemolytic anemia is caused
by IgM antibody, only the test for complement
components would be positive.
DIAGNOSIS
B. The indirect Coombs’ test
● used in the CROSSMATCHING of blood to
prevent a transfusion reaction.
● it is used either to determine the presence of
a particular antibody in a patient or to type
patient red blood cells for specific blood group
antigens.
● In vitro → binding of antibody to red cells,
rather than in vivo binding, is detected by this
method.
● This is a two-step process, in which red blood
cells and antibody are allowed to combine at
37°C, and then the cells are carefully washed
to remove any unbound antibody.
● Antihuman globulin is added to cause a visible
reaction if antibody has been specifically
bound.
● Any negative tests → are confirmed by
quality-control cells, which are coated with
antibody
Type III Hypersensitivity
● Also called → immune complex mediated
hypersensitivity
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The mechanism involved → ANTIGEN ANTIBODY
COMPLEX - deposits on the tissues; pati yung
surrounding tissues kung saan naka-deposits yung
antigen antibody ay NASISIRA
● They are due to the deposition of Ag-Ab
complexes in tissues and bloodvessels
● These complexes → can destroy the
surrounding tissue directly or indirectly by
attracting neutrophils to the site of complex
deposition that release hydrolytic enzymes,
causing → local damage
● Sites in which this typically occurs include the
glomerular basement membrane, vascular
endothelium, joint linings, and pulmonary
alveolar membranes
→It involves immune complexes which are deposited
on different tissues and blood vessels
→ reaction SIMILAR to type 2 kasi LYSIS OF THE
ANTIGENS padin; involved in type 3 is IgG & IgM
TYPE 3 HYPERSENSITIVITY SYMPTOMS: ARTHRITIS -
pag na deposits sa joined kaya nagkakaroon ng
exaggerated immune response toward the antigen
which are deposited on JOINTS which also destroy the
surrounding tissues
ANTIGENS:
TYPE 1 - ALLERGEN
TYPE 2 - CELL SURFACE ANTIGEN
TYPE 3 - SOLUBLE ANTIGEN
CLINICAL MANIFESTATIONS: Type III
● The reaction may be general (serum sickness)
or may involve individual organs including skin
(systemic lupus erythematosus, arthus
reaction), kidney (lupus nephritis), lung
(aspergillosis), blood vessels (polyarteritis),
joints (rheumatoid arthritis) or other organs.
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● Vaccines and bee stings may also trigger this

● This reaction may be the pathogenic
mechanism of diseases caused → by many
microorganisms.
TYPE III: ARTHUS REACTION
type of reaction.
● Generalized symptoms appear in → 7 to 21
days after injection of the animal serum and
include headache, fever, nausea, vomiting,
joint pain, rashes, and lymphadenopathy.

TYPE III: Serum sickness

Systemic lupus erythematosus (SLE) and rheumatoid

arthritis are two such examples.

● In SLE, antibodies are directed against

constituents such as DNA and nucleohistones,
which are found in most cells of the body.
○ Immune complex deposition involves
multiple organs, but the main damage
occurs to the glomerular basement
membrane in the →kidney.

● In rheumatoid arthritis, an antibody called →

rheumatoid factor is directed against IgG.
○ Immune complex deposition occurs in
the membranes of → inflamed joints.
○ Complement enhances tissue
destruction in both diseases
ARTHUS REACTION - example of localized type 3
hypersensitivity
Testing for Type III Hypersensitivity
Type III Hypersensitivity
● In specific diseases such as SLE and
rheumatoid arthritis, the presence of antibody
can be detected by:

○ agglutination reactions using

antigen-coated carrier particles,
including red blood cells or latex
particles, or enzyme immunoassays.

● Fluorescent staining of tissue sections has also

been used to determine deposition of
immune complexes in the tissues.
TYPE III: Serum sickness
● A more general method of determining
● Serum sickness results from → passive immune complex diseases is by measuring
immunization with animal serum, usually complement levels.
horse or bovine, used to treat such infections
as diphtheria, tetanus, and gangrene.

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● Decreased levels of individual components or ➔ CONTACT ANTIGENS - environmental antigen

decreased functioning of the pathway may be that come into direct contact with skin causing
indicative of antigen–antibody combination. now →CONTACT DERMATITIS such as POISON
IVY, POISON OAK, METALS, NICKELS, LATEX,
Type IV Hypersensitivity HAIR DYES, & COSMETIC

● T cells → is the mediator; and the activity is → Pathogen that are commonly induce delayed
DELAYED usually more on→ 24hrs-72hrs hypersensitivity:
● A hypersensitivity reaction mediated by
sensitized T cells releasing → lymphokines, ● MYCOBACTERIUM TUBERCULOSIS
attracting → macrophage to the site and ● M LEPRAE
activating them. ● PNEUMOCYSTIS CARDINI
● Once the macrophage arrives, they begin to ● LEISHMANIA
cause tissue damage that may develop into a ● HERPES VIRUSES
→ chronic granulomatous reaction if antigen ● Allergic skin reactions to bacteria, viruses,
persists. fungi, and environmental antigens such as
● It is also called DELAYED type hypersensitivity → poison ivy typify this type of
since following secondary exposure to the hypersensitivity
offending antigen, the manifestations of the
interaction do not appear for → more than 24
hours.
● It is also known as cell mediated or delayed
type hypersensitivity.

ROBERT KOCH - first observed that individuals infected

with MYCOBACTERIUM TUBERCULOSIS

● The classical example of this hypersensitivity is

tuberculin (montoux) reaction which peaks
48hours after the injection of antigen (PPD or
old tuberculin). The lesion is characterized by
induration and erythema.
● Another delayed hypersensitivity is contact
dermatitis (poison ivy, chemicals, heavy TYPE IV: Contact Dermatitis
metals) in which the lesions are more popular
● The MOST common causes include poison ivy,
● Mechanism of damage in delayed poison oak, and poison sumac, all of which
hypersensitivity include T lymphocytes and release the chemical urushiol in the plant sap
monocytes and/or macrophages. and on the leaves.
● Cytokine → production by the Th1 cells causes
● Cytotoxic T cells (Tc) cause direct damage → macrophages to accumulate and release
whereas helper T (TH1) cells are the one that cytokines and other substances that produce
activate CD8 → secrete cytokines which → a local inflammatory response
activate cytotoxic T cells and recruit and
activate monocytes and macrophages, which ● It produces a skin eruption characterized by
cause the bulk of the damage erythema, swelling, and the formation of
papules that appears from 6 hours to several
The antigens that can trigger type 4 hypersensitivity: days after the exposure
➔ INTRACELLULAR PATHOGEN - bacteria, viruses
and fungi
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 IMMUNOLOGY AND SEROLOGY
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Skin Testing for Delayed Hypersensitivity

SUMMARY
● Major lymphokines → involved in delayed
hypersensitivity reaction include → monocyte
chemotactic factor, interleukin-2, interferon,
TNF.

● Diagnostic test in vivo include delayed

cutaneous reaction (montoux test) and patch
test (for contact dermatitis).

PATCH TEST FINAL EVALUATION: 120 HOURS

● In vitro tests for delayed hypersensitivity

include mitogenic response,
lympho-cytotoxicity and IL-2 production.

● Corticosteroids and other immunosuppressive

agents are used in treatment. HUMORAL MEDIATED CELL-MEDIATED
Drugs used in managing a type 4 hypersensitivity are HYPERSENSITIVITY HYPERSENSITIVITY
→CORTICOSTEROIDS REACTIONS REACTIONS

Includes Types I, II, III Include type IV reactions

and V reactions Reactions are delayed in
time

Immune reactions are Reactions are delayed in

observed minutes after time
antigen exposure

Inflammatory reactions Reactions are

are characterized by characterized by
more fluid and significant cell infiltraton
erythema (wheal and with resultant
flare reaction) induration
Hypersensitivity Reactions

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 IMMUNOLOGY AND SEROLOGY
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IMMUNODEFICIENCY DISORDER
Unit Intended Outcomes:
Identify and differentiate the different
immune-deficiency disorders in relation to their
general immunologic defects, defects in the B-cell,
T-cell, myeloid, or complement systems.
OVERVIEW
Immunodeficiencies is a collective name for a LARGE
GROUP OF DISORDERS → when the function of the
immune system is NOT strong enough to protect the
organism against foreign invaders, especially
microbes and their toxic products, or against the
organism’s own malignantly transformed cells or
what we know as → CANCER CELLS
IMMUNE DISORDERS → are a collective disorders in
which there is a part of immune system that is missing
or dysfunctional → that’s why nagkakaroon ng mga
dysfunction or deficient ang ating immune system
People with conditions they have tendency to have a
decreased ability to defend themselves against
difference infections or against different
microorganisms → that’s why the decreased ability to
protect themselves they are MORE SUSCEPTIBLE to
developing certain types of diseases or acquiring
certain types of infection as well as developing certain
types of CANCER
These is a RISK FACTORS that could make those
patients more SUSCEPTIBLE to develop → more
MALIGNANCIES/CANCER CELLS
The clinical symptoms associated
with immunodeficiencies range from → very MILD or
subclinical to → SEVERE MANIFESTATIONS, and the
MOST clinical symptom of patients having
immunodeficiency is having → RECURRENT
INFECTIONS or FAILURE TO THRIVE or FAILURE of
eliciting an immune response or immune mechanism
against different invasion of microorganisms
→ so usually ang pinaka SYMPTOM na meron
immune deficiency disorder pag paulit ulit yung
INFECTION because sa mga taong may
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IMMUNODEFICIENCY DISORDER or
IMMUNOCOMPROMISED PEOPLE yung mga simpleng
sakit na NORMALLY na nalalabanan ng katawan.
But for immunodeficiency patients hindi nila kayang
labanan nagiging susceptible sila that’s why the MOST
CLINICAL SYMPTOMS OF IMMUNODEFICIENCY
DISORDERS are → RECURRENT INFECTIONS because
of → DYSFUNCTION OF IMMUNE SYSTEM
Immunodeficiencies can be INHERITED or ACQUIRED
INHERITED - known as PID (Primary immune
deficiencies)
→ PRIMARY IMMUNE DEFICIENCY DISORDER
ACQUIRED - yung mga nakukuha such as HIV which
causes AIDS which is an immunodeficiency disorder is
ACQUIRED like nakukuha sa infection or
microorganism,
MALIGNANCIES, IMMUNOSUPPRESSIVE THERAPIES
and that known as
→ SECONDARY IMMUNE DEFICIENCY DISORDERS
Categories of Primary Immunodeficiencies
Category 1: Combined Immunodeficiencies
Category2: Combined Immunodeficiencies with
associated or Syndromic Features
Category 3: Predominantly Antibody Deficiencies
(pinakamarami) - the PREDOMINANTLY antibody
deficiencies have the MOST common
immunodeficiency representing about 50% of all the
PIDs
Category 4: Diseases of Immune Dysregulation
Category 5: Congenital Defects of Phagocyte Number,
Function, or Both
Category 6: Defects in Innate Immunity
Category 7: Auto-inflammatory Disorders
Category 8: Complement Deficiencies
Category9: Phenocopies of Primary
Immunodeficiencies
RECURRENT INFECTION - most common hallmark of
immunodeficiency
the most common hallmark na meron talagang or
immunodeficiency disorder yung isang patient is the
→ RECURRENCE OF REPEATED INFECTION and even
by organism considered as → LOW VIRULENT
 IMMUNOLOGY AND SEROLOGY
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IMMUNOCOMPETENT/NORMAL IMMUNE SYSTEM -
meron mga organism that DO NOT cause severe
infections to those people who have normal immune
response
LOW VIRULENT ORGANISM - these organism DO NOT
normally cause severe infections in human
BUT in patient with IMMUNODEFICIENCY DISORDER
nagkaroon sila ng → REPEATED RECURRENT with
that LOW VIRULENT ORGANISMS and nagiging severe
pa
In the PAST IMMUNODEFICIENCY DISORDER have
been classified as defects in:
➔ T cells
➔ B cells
➔ Phagocytes
➔ Complement and other component of INNATE
BUT in 2014 the international union of immunological
societies have updated their classification of the
PRIMARY IMMUNODEFICIENCIES by grouping them
into 9 different categories based on their
characteristics, clinical features, immunologic defects
and genetic abnormalities kung ano ba yung na
inherent mutation or AUTOSOMAL/RECESSIVE GENES
→ that would cause the immune deficiency disorder
Category 1: Combined Immunodeficiencies
CATEGORY 1 - known as the → COMBINED
IMMUNODEFICIENCY because deficiency or disorder
under category 1 involves defect in both HUMORAL
and those MEDIATED BY B CELLS as well as defects in T
CELLS functions.
● Defects in both humoral (B cell) and
cell-mediated (T cell) immunity
● Result from → MUTATION that affect
development of both types of →
lymphocytes (which are B CELLS and T CELLS
may defect pareho and T Cells and B Cells
function that’s why DECREASED and
DYSFUNCTIONAL) or cause defective
interaction between the two antigen-specific
limbs of the adaptive immune system.
→ a SEVERE DEFECT on T Cells function will also have
effects on the IMMUNOGLOBULIN LEVELS as well
These categories involves defects on
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77
→ B Cells and T Cells (kasi interrelated sila pareho)
Diseases include:
○ Severe Combined Immunodeficiency (SCID)
○ Purine-Nucleoside Phosphorylase (PNP) Deficiency
Severe Combined Immunodeficiency Disease (SCID)
● MOST SERIOUS of the among congenital
immunodeficiencies meaning there is →
ASSOCIATION or there is an INVOLVEMENT OF
A MUTATION in the GENETIC COMPOSITION/
INHERITED GENES
➔ POTENTIALLY FATAL/MOST SERIOUS
➔ PRIMARY IMMUNODEFICIENCY
➔ Combined absence of T Cells and B cells
● A group of related diseases that all affect → T-
and B-cell function but with differing causes
causing a SEVERE COMBINED IMMUNODEFICIENCY
DISEASE wherein parehong T Cells and B cells
DEPLETED FUNCTION or DECREASE FUNCTION
Interleukin-2 receptor gamma (IL2RG)
Usually the MUTATION occurs with the frequency of
about 1 in 50,000 births - NOT common, VERY RARE
immunodeficiency and usually cause in → mutation
INTERLEUKIN-2 RECEPTOR GAMMA GENES (IL2RG)
located on the X-chromosome is the most common
form of the disease
INTERLEUKIN-2 RECEPTOR GAMMA GENES - is a gene
that CODES for a PROTEIN CHAIN called the COMMON
GAMMA CHAIN that is common to receptor for
INTERLEUKIN 2,7,9,15,21 or those INTERLEUKINS that
are secreted by T CELLS
NORMAL SIGNALING CANNOT occur in cells with the
effective receptors and thus halting the maturation of
T cells and B cells.
If meron MUTATION INTERLEUKIN-2 RECEPTOR
GAMMA GENES (kung wala or if meron mutation sa
INTERLEUKIN-2 RECEPTOR GAMMA GENES -
nagkakaroon ng DEFECTIVE SIGNALING and HALTING
now the natural maturation of T cells and B cells
 IMMUNOLOGY AND SEROLOGY
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that's why nagkakaroon ng DEPLETED or DECREASED T
Cells and B Cells function.
JAK3 gene
○ An additional defect in the JAK3 gene
JACK3 GENE - is required for processing an
INTERLEUKIN BINDING SIGNAL from the cells
membrane will enter the nucleus and ACTIVATE
NORMAL SIGNALING and DIFFERENTIATION of T Cells
and B Cells
Adenosine deaminase (ADA) deficiency
○ About 15% to 20% of the patients with SCID
have an adenosine deaminase (ADA)
deficiency, leading to a → NEGATIVE T-B-NK-
phenotype - meaning nagkakaroon ng SEVERE
DEPLETION or PROGRESSIVE decrease in
lymphocyte numbers.
ADA DEFICIENCY - affects an enzyme that is
involved in the METABOLISM of PURINES
(toxic metabolites of purines accumulate in
LYMPHOID CELLS that’s why because of these
toxic metabolites of purines due to the
deficiency of ADA - nagkakaroon ngayon ng
IMPARED PROLIFERATION of T Cells and B
Cells) that’s why can also result in SCID →
leading to the progressive DECREASE
LYMPHOCYTE number also leading to a
PHENOTYPE which causes REDUCE or
progressive DECREASE LYMPHOCYTE number
- this is SIMILAR to another form APAID
deficiency which is the PNP
In patients with only MILDLY REDUCED ADA ACTIVITY
so meron lang SLIGHT IMPAIRMENT of immune
function → so it DEPENDS upon the SEVERITY of the
ADA activity
RAG-1 or RAG-2 GENES
○ A MUTATION in a recombinase activating
gene (RAG-1 or RAG-2) - important because
they are activating genes that are very
important in DIFFERENTIATION of PRO-B Cells
or IMMATURE B cells
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Patient with SCID were found to have a MUTATION in
RAG-1 or RAG-2 → they LACK both T Cells and B Cells
BUT they have functioning NK cells
NK cells are NOT affected
RAG-1 or RAG-2 GENES
- are important in differentiation of the EARLY
CELLS or EARLY PRO-B Cells and PRE-B Cells
stages
- very important in the REARRANGEMENT of
the light chains and heavy chains genes
- aslo for the REARRANGEMENT of the
DEOXYRIBONUCLEIC ACID (DNA) inside of B
Cells
- necessary to PRODUCE to FUNCTIONAL
IMMUNOGLOBULINS in the differentiation of
B cells during the coding
- functional T Cells receptor or TCR
- important in the normal maturation and
differentiation of T Cells and B Cells
→ that’s why a DEFECT or MUTATION of these genes
could also result in SEVERE COMBINED DEFICIENCY of
T Cells and B Cells function
→ usually with patients with RAG-1 or RAG-2
DEFICIENCY have DECREASED CLASS 2 MHC(antigen
presentation) MOLECULE adding up to the deficiency
or to the disorder of the immune system.
CD45
○ A molecular defect in the gene encoding a
common leukocyte protein called CD45
CD45 - transmembrane phosphatase which regulate
the SIGNAL TRANSDUCTION of T Cells and B Cells
receptors.
→ that’s why a DEFECT of these PROTEIN can also
IMPAIR of immune response
→ all of these CONFORMATIONAL or GENETIC
ABNORMALITIES can induce an immune deficiency
disorder, such as this SCID
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● Patients with SCID generally present early in
infancy with infection by nearly any type of
organism.
● Oral candidal yeast infections, pneumonia,
and diarrhea are the most common
MANIFESTATION/SYMPTOMS.
● The administration of live vaccines can cause
severe illness.
● Unless immune reconstitution can be
achieved by bone marrow transplantation or
by specifically replacing a deficient enzyme,
patients with SCID die before they are → also have neurological disorders (MENTAL
2 years old
successful reconstitution by using → STEM CELLS so
that mag increase yung number ng T Cells and B Cells
VERY FATAL & VERY SERIOUS PID AMONG CHILDREN →
the severe combine kasi parehong T Cells and B Cells
are severely DECREASED that’s why they are very
prone and usually ang kinamamatay nila is
SECONDARY INFECTIONS kasi hindi malabanan ang
kanilang katawan since bata pa sila and common is
PNEUMONIA
Purine-Nucleoside Phosphorylase (PNP) Deficiency
also a CATEGORY 1 in which there is combined
immunodeficiency of T Cells and B Cells and the
condition present in INFANCY as well with RECURRENT
● CATEGORY 1 in which there is combined
immunodeficiency of T Cells and B Cells The
condition presents in INFANCY with
RECURRENT or chronic pulmonary infections,
oral or cutaneous candidiasis, diarrhea, skin
infections, urinary tract infections, and
FAILURE to thrive or failure to produce
immune response.
● PNP deficiency is also a RARE AUTOSOMAL
RECESSIVE TRAIT and affects an enzyme
involved in the → metabolism of purines
(same as ADA)
● Produces a MODERATE to SEVERE defect in
cell mediated immunity that’s why
→ The number of T cells progressively
decreases
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79
● The levels of immunoglobulins are generally
normal or increased BUT the T cells
progressively decreases → because of the
accumulation of deoxyguanosine
triphosphate, a toxic purine metabolite.
PNP deficiency - lacking an enzyme that is involved in
purine metabolism lead to TOXIC METABOLITE OF
PURINE known as → deoxyguanosine triphosphate
● About two thirds of PNP-deficient patients
DISABILITIES), but NO characteristic physical
abnormalities
PNP deficiency can be confused with
neonatal HIV infection. The two conditions can
usually be distinguished by specific tests for HIV and
by assays for PNP activity.
Category 2: Combined Immunodeficiencies With
Associated or Syndromic Features
Category 2 are characterized by NON IMMUNOLOGIC
features in addition to the combined
immunodeficiency.
→ known as combined immunodeficiencies with
associated or SYNDROMIC FEATURES
● Diseases in this category are typically caused
by → DEFECTS in CELL-MEDIATED IMMUNITY
● Often these diseases can result from
abnormalities at different stages of T-cell
development → LEADS to other problems
with other branches of immune response such
as HUMORAL IMMUNITY
Examples of specific immunodeficiencies in this
category are:
○ Wiskott-Aldrich syndrome (WAS)
○ DiGeorge anomaly
○ Ataxia-Telangiectasia (AT)
Wiskott-Aldrich Syndrome (WAS)
AUTOSOMAL DISORDER & NON IMMUNOLOGIC
● A rare X-linked recessive syndrome that is
defined by the triad of immunodeficiency,
eczema, and thrombocytopenia
WAS is usually lethal in childhood because of
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→ RECURRENT infection, hemorrhage because of
thrombocytopenia, or malignancy/formations of
cancer.
The laboratory features of WAS include a decrease in
platelet number and size with a prolonged bleeding
time because of the decrease in platelet function.
The bone marrow contains a normal or somewhat
increased number of megakaryocytes.
● Milder variants have also been described such
as an → X-linked form of thrombocytopenia
● There are abnormalities in both the
cellular (t cells) and humoral (b cells)
branches of the immune system related to a
general defect in antigen processing
As a result, patients display a severe deficiency of the
naturally occurring IgM antibodies to ABO blood
group antigens known as → ISOHEMAGGLUTININS
● Absence of isohemagglutinins is the → most
consistent laboratory finding in WAS and is
often used diagnostically
● Patients also have persistently increased
levels of →serum alpha fetoprotein
Patients with WAS can have a variety of different
patterns of immunoglobulin levels, but they usually
have low levels of IgM, normal levels of IgA and IgG,
and increased levels of IgE.
● The primary molecular defect in the syndrome
appears to be an abnormality of the integral
membrane protein CD43
CD43 - which is involved in the regulation of protein
glycosylation
Abnormalities cause defective actin polymerization
and affect its signal transduction in lymphocytes and
platelets
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80
● Platelets have a shortened half-life and T
lymphocytes are also affected, although B
lymphocytes appear to function normally
Current treatment: Transplantation of bone marrow
or cord blood stem cells from an HLA identical sibling.
Splenectomy can be very valuable in controlling the
thrombocytopenia.
DiGeorge Anomaly
known as CONGENITAL THYMIC HYPOPLASIA
● A developmental abnormality of the third and
fourth pharyngeal pouches that → affects
thymus development in the embryo
→ All organs derived from these embryonic structures
can be affected
- Primary Immunodeficiency disease caused by
ABNORMAL MIGRATION and development of
certain cells and tissues during the fetal
development
Defects of the third and fourth pharyngeal pouches
during the embryogenesis and therefore affects the
development of the THYMUS
These children tend to have severe, recurrent viral
and fungal infections.
.
● A disorder associated by a defect in
chromosome 22
● A severe and persistent DECREASE in T-cell
numbers and other branches of immune
response will also be INDIRECTLY AFFECTED
such as HUMORAL IMMUNE RESPONSE
● Patients tend to have severe, recurrent viral
and fungal infections.
● Severely affected children usually present in
the neonatal period with TETANY (caused by
hypocalcemia resulting from
hypoparathyroidism) or manifestations of
cardiac defects
Non-Immunologic features are
→ MENTAL RETARDATION (mental disorder)
➔ mental retardation
➔ absence of ossification of the hyoid bone
➔ cardiac anomalies
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➔ abnormal facial development (FISH SHAPE
MOUNTH & tendency to have a LOW EARS)
➔ thymic hypoplasia - kasi yung problem is may
defect sa THYMUS DEVELOPMENT which
IMPAIRS the T cells number)
Many patients with a partial DiGeorge anomaly have
only a minimal thymic defect and, thus, near normal
immune function. However, about 20% of children
with a defect of the third and fourth pharyngeal
pouches have a severe and persistent decrease in
T-cell numbers.
The immunodeficiency associated with the DiGeorge
anomaly is a QUANTITATIVE DEFECT in thymocytes.
→ number yung cause, there is SEVERE or PERSISTENT
DECREASE in T cells number.
NOT enough to become activated.
● The only effective therapy for AT is allogeneic
bone marrow transplantation
Usually the cause of Ataxia-Telangiectasia (AT) is a
AUTOSOMAL RECESSIVE DEFECT IN THE AT GENE
→ results in a defective kinase involved in DNA repair
and in cell cycle control
Abnormal rearrangement of TCR and immunoglobulin
genes does not occur normally. Patients’ lymphocytes
often exhibit chromosomal breaks and other
abnormalities involving the TCR genes in T cells and
immunoglobulin genes in B cells.

Treatment: Fetal thymus transplantation, Bone The levels of IgG2, IgA, and IgE are often low or
marrow transplantation, administration of thymic absent, although the pattern can be quite variable.
hormones In addition, the number of circulating T cells is often
decreased. Death usually occurs in early adult life
Ataxia-Telangiectasia (AT) from either pulmonary disease or malignancy
● A RARE autosomal recessive syndrome
characterized by cerebellar ataxia and
telangiectasias
Cerebellar ataxia - involuntary muscle movements
Telangiectasias - capillary swelling resulting in red
blotches on the skin especially on the earlobes and
conjunctiva.

→ Blood vessels in the sclera of the eyes may be

dilated
and there may also be a reddish butterfly area on the
face and ears.
● 95% of patients exhibit increased levels of
serum alpha-fetoprotein
→ The incidence of this disease is between 1:10,000
to 1:100,000, although as much as 1% of the
population is heterozygous for the gene

→ Antibody response to antigens, especially

Category 3: Predominantly Antibody Deficiencies
polysaccharides, are usually slow/blunted.
● Include genetic defects in B-cell maturation or
mutations leading to defective interactions
● Abnormal genes produce a combined
between B and T cells
→ defect of both humoral and cellular
This category encompasses conditions in which the
immunity
MAIN characteristic is → low levels of serum
● There is a defect in the AT gene is located on
immunoglobulins.
chromosome 11, region q22

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deficiency in immunoglobulin is
→ AGAMMAGLOBULINEMIA or (meron deficiency in
the concentration of antibodies)
The mechanisms of the AGAMMAGLOBULINEMIA
include genetic defects in B-cell maturation or
mutations leading to defective interactions between B
and T cells.
Wide range of immunoglobulin deficiency states
have been reported and involve virtually all
combinations of immunoglobulins and all degrees of
severity. In some cases, only a single isotype of one
immunoglobulin class is deficient, whereas all of the
other isotypes are NORMAL.
In evaluating immunoglobulin deficiency states, it is
important to remember that blood levels of
immunoglobulins change with age.
The blood level of IgG at birth is about the SAME as
the adult level, reflecting transfer of maternal IgG
across the placenta.
→ The IgG level declines over the first 6 months of life
as maternal antibody is catabolized.
→ Levels of IgA and IgM are very low at birth. The
concentrations of all immunoglobulins gradually RISE
when the infant begins to produce antibodies at a few
months of age in response to environmental stimuli.
IgM reaches normal adult levels first, around 1 year of
age, followed by IgG at about 5 to 6 years of age.
Predominantly Antibody Deficiencies include:
○ Transient hypogammaglobulinemia of infancy
○ Selective IgA deficiency
○ Btk deficiency
○ Common variable immunodeficiency
○ Isolated IgG subclass deficiency
○ CD154 deficiency
Transient Hypogammaglobulinemia of Infancy with
Normal Numbers of B Cells
All infants experience low levels of immunoglobulins
at approximately 5 to 6 months of age. However, in
some babies the low levels persist for a longer time.
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82
● A relatively common primary
immunodeficiency disease that affects
infants and young children.
● Infants experience low levels of
immunoglobulins persisting for a longer
period of time.
→ Because these children do NOT begin synthesizing
immunoglobulins promptly, they can experience
severe pyogenic sinopulmonary and skin infections as
protective maternal IgG is cleared.
● Cell-mediated immunity is NORMAL and there
may be normal levels of IgA and IgM
● IgG appears to be the → MOST AFFECTED
dropping to at least 2 standard deviations
(SDs) below the age-adjusted mean with or
without a depression of IgM and IgA
● Immunoglobulin levels in infants with this
condition usually NORMALIZE spontaneously,
often by 9 to 15 months of age.
The cause may be related to a delayed maturation
of one or more components of the immune system,
possibly Th cells.
X-Linked Bruton’s Tyrosine Kinase (Btk) Deficiency
→ first described in 1952, is X chromosome linked, so
this syndrome affects MALES almost exclusively
● A hereditary immunodeficiency due to the
failure of Blymphocytes to mature and
rearrangement failure of Ig Heavy Chain
● Patients with X-linked agammaglobulinemia
lack of circulating mature CD19+ B cells and
exhibit a deficiency or lack of
immunoglobulins of all classes
→ They have NO plasma cells in their lymphoid
tissues.
The patients do, however, have pre-B cells in their
bone marrow. Because of the lack of B cells, the
tonsils and adenoids are →small or entirely ABSENT
and lymph nodes lack normal germinal centers.
They develop recurrent bacterial infections beginning
in INFANCY as maternal antibody is cleared. The
patients most commonly develop
→ sinopulmonary infections caused by encapsulated
organisms such as streptococci, meningococci, and
Haemophilus influenzae.
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→ Other infections seen include bacterial otitis
media, bronchitis, pneumonia, meningitis, and
dermatitis.
X-linked hypogammaglobulinemia results from
arrested differentiation at the pre–B-cell stage,
leading to a → complete absence of B cells and
plasma cells.
The underlying genetic mechanism is a deficiency of
an enzyme called the Btk in B-cell progenitor cells.
Lack of the enzyme apparently causes a failure of
immunoglobulin VH gene rearrangement.
The syndrome can be differentiated from transient
hypogammaglobulinemia of infancy by the absence of
CD19+ B cells in the peripheral blood, the abnormal
histology of lymphoid tissues, and its persistence
beyond 2 years of age.
● T cells are normal in number and function.
TREATMENT: administration of intramuscular or
intravenous immunoglobulin preparations and
vigorous antimicrobial treatment of infections
Selective IgA Deficiency
● It is hypothesized that lack of IgA is caused by
impaired differentiation of lymphocytes to
become → IgA-producing plasma cells
Selective IgA deficiency is the most common
congenital immunodeficiency, occurring in about 1 in
500 persons. Most patients with a deficiency of IgA
are ASYMPTOMATIC. Those with symptoms usually
have infections of the respiratory and gastrointestinal
tract and an increased tendency to develop
autoimmune diseases such as:
➔ systemic lupus erythematosus (SLE),
➔ rheumatoid arthritis (RA)
➔ celiac disease
➔ thyroiditis
About 20% of the IgA-deficient patients who develop
infections also have an IgG2 subclass deficiency.
● IgE antibodies specifically directed against IgA
are produced by 30% to 40% of patients with
severe IgA deficiency, causing anaphylactic
reactions when blood products containing IgA
are transfused
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83
Patients with severe IgA deficiency have no other
symptoms, the IgA deficiency may not be detected
until the patient experiences a transfusion reaction,
resulting in the production of anti-IgA antibodies.
Therefore, products for transfusion to known
IgA-deficient patients should be collected from
IgAdeficient donors or cellular products should be
washed to remove as much donor plasma as possible.
DEPLETED is IgA type of immunoglobulins.
→ Usually the reaction is ANAPHYLACTIC during
BLOOD TRANSFUSION
Common Variable Immunodeficiency (CVI)
● A low incidence, but the most common PID
with a severe clinical syndrome
● The disorder can be congenital or acquired, or
familial or sporadic, and it occurs with equal
frequency in men and women.
CVI - heterogeneous group of disorders with a
prevalence of about 1 in 25,000.
→ Patients usually begin to have symptoms in their
20s and 30s, but age at onset ranges from 7 to 71
years of age.
CVI is characterized by hypogammaglobulinemia that
leads to recurrent bacterial infections, particularly
sinusitis and pneumonia.
● There is usually a deficiency of both IgA and
IgG, but selective IgG deficiency may occur
● It is also associated with an increased risk of
lymphoproliferative disorders, gastric
carcinomas, and autoimmune disorders
● Diagnosed by demonstrating a low serum IgG
level in patients with recurrent bacterial
infections
up to 20% of CVI patients develop HERPES ZOSTER
(shingles), a much higher incidence than in
immunologically normal young adults.
CVI is often associated with a spruelike syndrome
characterized by malabsorption and diarrhea.
The most common autoimmune manifestations of CVI
are immune thrombocytopenia and autoimmune
hemolytic anemia.
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Other symptoms may include:
➔ lymphadenopathy
➔ splenomegaly
➔ intestinal hyperplasia
Blood group isohemagglutinins, or the so-called
naturally occurring antibodies, are typically absent or
low. In contrast to X-linked agammaglobulinemia,
most patients with CVI have normal numbers of
mature B cells. However, these B cells DO NOT
differentiate normally into immunoglobulin-producing
plasma cells.
COMPENSATED by the HIGH AMOUNTS kasi normal
naman si IgG2 and IgG4
Most IgG antibodies directed against protein antigens
are of the IgG1 and IgG3 sub-classes, whereas most
- IgG antibodies against carbohydrate antigens
are IgG2 or IgG4.
- Thus, deficiencies involving IgG1 or IgG3 lead
to a reduced capability of responding to
protein antigens such as toxins, whereas
selective deficiencies of IgG2 can result in
impaired responses to polysaccharide
antigens, which cause recurrent infections
with polysaccharide-encapsulated bacteria
such as:
➔ Streptococcus pneumoniae
➔ H influenzae.

Three major types of cellular defects have been

found in CVI patients: IgG4 - The most common subclass deficiency, although
➔ T cells or their products appear to suppress IgG4 subclass deficiency may have the least clinical
differentiation of B cells into plasma cells. significance (NOT SEVERE )
➔ T cells may FAIL to provide adequate help to
support terminal differentiation of B cells. IgG1 & IgG3 - TOXIN, bacteria produce ENTEROTOXIN
➔ Primary defect in the B-cell line in some IgG2- can result in impaired
patients. polysaccharide-encapsulated bacteria such as
Hindi siya AGAMMAGLOBULINEMIA meron plasma Streptococcus pneumoniae & H influenzae.
cells and B cells BUT hindi siya mag differentiate
normally into PLASMA CELLS → that’s why IMPAIRED IgG1 deficiency - being the least common
ang ADAPTIVE IMMUNITY with patients with CVI
● IgG1 and IgG3 sub-classes: protein antigens
● CVI can usually be effectively treated with ● IgG2 or IgG4 sub-classes: carbohydrate
intramuscular or intravenous immunoglobulin antigens
preparations ● The most common subclass deficiency is IgG4,
with IgG1 deficiency being the least common
Isolated IgG Subclass Deficiency
This may result in a LOW LEVEL of IgG
● These are conditions where the level(s) of one
or more of the four IgG subclasses is (are)
more than two SDs below the mean
age-appropriate level
Normally, about 70% of the total IgG is IgG1, 20%
IgG2, 6% IgG3, and 4% IgG4.

In patients with recurrent infections, levels of the

different subclasses should be measured if the total
IgG level is normal kasi pwede ang baba ni IgG1 and
IgG3 pero NORMAL padin yung total IgG → because
the LOW NUMBER of IgG1 AND IgG3 is being

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Category 4: Diseases of Immune Dysregulation
MAJORITY AUTOIMMUNE DISEASE
● Many of the diseases may involve normal
numbers of T or B cells but with reduced
control over their functions
The autoimmune lymphoproliferative syndrome
(ALPS)
→ for example, may involve mutations in genes coding
for caspase enzymes involved in APOPTOSIS of B Cells
● May also have features of autoimmunity
● Autoimmune lymphoproliferative syndrome
(ALPS)
● CD25 deficiency
CD25 deficiency - is manifested by a lack of T
regulatory (Treg) cells, which leads to
lymphoproliferation and autoimmunity
● Mutations in the FoxP3 gene
Mutations in the FoxP3 gene - which is required for
Treg differentiation, may show a similar clinical
presentation
● Chediak-Higashi syndrome: caused by a
mutation in the LYST gene
Chediak-Higashi syndrome - an immunodeficiency
with hypopigmentation (loss of skin color) caused by
a mutation in the LYST gene, is characterized by a
reduced number of natural killer (NK) cells and
neutrophils, as well as an increased production of
inflammatory proteins.
Patients with Chediak-Higashi syndrome show
granulocytic inclusions attributed to enlarged
lysosomes
Category 5: Congenital Defects of Phagocyte Number,
Function, or Both
DEPLETED neutrophils and phagocytic cells or
NORMAL naman yung count pero DEFECTIVE naman
sila → hindi kaya yung MICROBIAL KILLING that’s why
impaired din ang immune response
➔ Defective ang number of phagocytes
➔ or NORMAL ang NUMBER but DEFECTIVE yung
function or combination of BOTH
➔ ABNORMAL FUNCTION and LOW NUMBER of
PHAGOCYTES
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85
Characterized by abnormalities in phagocytic cells
○ Chronic Granulomatous Disease (CGD)
○ Other Microbiocidal Defects
○ Leukocyte Adhesion Deficiency (LAD)
Neutrophils play a crucial role in the immediate and
nonspecific response to invading organisms by
responding before specific antibody and cell-mediated
immune responses can be mounted.
- Neutrophils are even more effective at
ingesting and killing organisms coated with
specific antibody and thus continue to play an
important role in host defense even after an
adaptive immune response is established.
- To destroy invading organisms, neutrophils
must adhere to vascular endothelial lining
cells, migrate through the capillary wall to a
site of infection, and ingest and kill the
microbes
Defects affecting each of these steps can lead to an
increased susceptibility to pyogenic infections.
Chronic Granulomatous Disease (CGD)
● A group of disorders involving inheritance of
either an X-linked or autosomal recessive gene
that affects neutrophil microbiocidal function
CGD - is the one of the most common and best
characterized of neutrophil abnormalities
● Result in the inability of the patient’s
neutrophils to produce the reactive forms of
oxygen necessary for normal bacterial killing
➔ Normally, neutrophil stimulation leads to the
production of reactive oxygen molecules, such
as hydrogen peroxide (H2O2), by NADPH
oxidase reactivity on the plasma membrane.
➔ The plasma membrane enfolds an organism as
it is phagocytized and hydrogen peroxide is
generated in close proximity to the target
microbe.
➔ Neutrophil granules fuse with and release the
enzyme myeloperoxidase into the forming
phagosome. The myeloperoxidase uses the
hydrogen peroxide to generate the potent
microbicidal agent, hypochlorous acid
Hexose monophosphate shunt - the process of
generating partially reduced forms of oxygen by
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stimulated neutrophils was first detected as an
increase in oxygen consumption, known as
→ RESPIRATORY BURST / OXIDATIVE BURST
NADPH oxidase system can result in the CGD
phenotype by making the neutrophil incapable of
generating an oxidative burst
● Symptoms include: recurrent suppurative
infections, pneumonia, osteomyelitis,
draining adenopathy, liver abscesses,
dermatitis, and hypergammaglobulinemia
● Staphylococcus aureus, Burk olderia cepacia,
and Chromobacterium violaceum; In patient
with CGD in additional to FUNGI such as
Aspergillus and Nocardia and INFECTION
usually begins before 1 year of age and the
syndrome is usually often FATAL in
CHILDHOOD for CGD
UNABLE to produce NORMAL BACTERIAL KILLING
because they are INCAPABLE of producing ROS or
activate form reactive oxygen species which are
necessary for normal bacterial killing kasi nga
→ ABNORMAL or INCAPABLE of generating an
OXIDATIVE BURST there is GENETIC DEFECT in the
several NADPH OXIDASE SYSTEM
➔ CGD was historically diagnosed by measuring
the ability of a patient’s neutrophils to reduce
the dye nitroblue tetrazolium (NBT).
➔ NBT reduction is caused by the production of
hydrogen peroxide and other reactive forms
of oxygen.
➔ Reduction converts the nearly colorless NBT
into a blue precipitate that can be assessed
visually on a microscope slide.
➔ DHR will fluoresce when it is reduced.
➔ The neutrophils are then activated using
phorbol myristate acetate (PMA), which is
mitogenic for neutrophils.
➔ The resultant oxidative burst will reduce the
DHR, resulting in fluorescence that may be
quantitated on a flow cytometer.
➔ Neutrophils from CGD patients will be unable
to undergo the oxidative burst and show less
fluorescence than normal neutrophils.
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86
➔ Although therapy with granulocyte
transfusions may allow resolution of an acute
infectious episode, it is impossible to provide
enough granulocytes to treat the chronic
condition.
➔ Administration of cytokines, such as
interferon, may increase the oxidative burst
activity in some patients. Continuous use of
antibiotics can greatly reduce the occurrence
of severe infections.
➔ Bone marrow transplantation or use of
peripheral blood stem cells may result in a
permanent cure
Diagnosis of Phagocytic Cells
● The nitroblue tetrazolium test (NBT) usually is
used to assay the phagocytic function of
neutrophils.
● This assay for metabolism and generation of
toxic molecules determines whether
phagocytic cells are using hexose
monophosphate shunt (HMP) and generating
toxic materials to kill microorganism.
● It is used in the diagnosis of chronic
granulomatous disease (CGD).
● The neutrophils of CGD patients fail to reduce
the NBT dye
● More recently, a flow cytometric assay has
been used. In this assay, neutrophils are
labeled with dihydrorhodamine (DHR).
● DHR will fluoresce when it is reduced.
● Uses Phorbol myristate acetate (PMA), which
is mitogenic for neutrophils.
● Neutrophils from CGD patients will be unable
to undergo the oxidative burst and will show
less fluorescence than normal neutrophils.
● This technique is more objective and
quantitative than the traditional NBT
technique
OTHER MICROBICIDAL DEFECTS
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Neutrophil glucose-6-phosphate dehydrogenase
deficiency
○ Aerobic system of neutrophils are impaired.
Neutrophil glucose-6-phosphate dehydrogenase
deficiency
- leads to an inability to generate enough
NADPH to supply reducing equivalents to the
NADPH oxidase system
- There is a HEMOLYTIC ANEMIA that is present
.because G6PD also causes POIKILOCYTOSIS
or ANOMALY in RBCs wherein nagkakaroon ng
HEINZ BODIES or PITTED GOLF BALL
appearance ang rbcs that's why they are being
destroyed in SPLEEN and nagkakaroon ng
HEMOLYTIC ANEMIA in G6PD
Myeloperoxidase deficiency
- These are SPECIFIC GRANULES in
NEUTROPHILS that are needed for bacterial
killing.
Myeloperoxidase deficiency is relatively common,
occurring in about 1 in 3,000 persons in the United
States.
○ Deficient patients may have recurrent
candidal infections
○ Defects of neutrophil secondary granules
- Defects of neutrophil secondary granules have
been described also.
However, the molecular nature of the defects is
unknown
Leukocyte Adhesion Deficiency (LAD)
● There is a deficiency in a protein called CD18,
which is a component of adhesion receptors
on neutrophils and monocytes (CD11b or
CD11c) and on T cells (CD11a)
● Abnormal adhesion, motility, aggregation,
chemotaxis, and endocytosis by the affected
leukocytes.
Even if microbiocidal activity is normal, neutrophils
cannot perform their functions properly if they fail to
leave the vasculature and migrate to a site of incipient
infection.
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87
Adhesion receptors on leukocytes and their counter
receptors on endothelial cells and the extracellular
matrix play important roles in these activities.
● Manifestations: delayed wound healing,
chronic skin infections, intestinal and
respiratory tract infections, and periodontitis
Category 6: Defects in Innate Immunity
Category 6 DEFECTS in the innate immune system. At
least one disease, chronic mucocutaneous
candidiasis, which was previously classified as a T-cell
defect, is now included in this classification
Other rare entities classified under this heading
include mutations in Toll-like receptors (TLRs)
● Chronic mucocutaneous candidiasis
● Other rare entities classified under this
heading include mutations in Toll-like
receptors (TLRs)
● Few clinical laboratory assays are currently
available for assessing innate immune system
functional capabilities
- defects can lead to bacterial infections
Category 7: Autoinflammatory Disorders
Autoinflammatory disorders are subdivided into two
classifications:
➔ Those involving the inflammasome
➔ non inflammasome conditions
Inflammasome - is a protein oligomer that contains
caspase enzymes and other proteins associated with
apoptosis.
- The inflammasome is located primarily in
myeloid cells and may be activated by various
microbial substances. Once activated, the
inflammasome stimulates the production of
the proinflammatory cytokines IL-1 and IL-18
● Genetic defects involving the inflammasome
include the Hyper IgD syndrome, also referred
to as periodic fever syndrome (mevalonate
kinase), and Muckle-Wells syndrome
(cryopyrin)
Muckle-Wells syndrome is caused by a mutation in
the CIAS1 gene coding for cryopyrin, a component of
the inflammasome.
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● Defects not involving the inflammasome
include tumor necrosis factor (TNF)
receptor-associated periodic syndrome
(TRAPS) (TNFRSF1A gene) -
TRAPS is caused by a mutation in the TNFRSF1A gene,
which codes for a TNF receptor and may result in
recurrent fevers, as well as ocular and joint
inflammation.
early-onset inflammatory bowel disease
(IBD) (IL-10 or its receptor)
Early-onset IBD - is caused by mutations in genes
coding for IL-10 or its receptor
Category 8: Complement Deficiencies
● Deficiencies in the early complement
components, C1q, C4, and C2, are usually
associated with a lupuslike syndrome
Complement - consists of a SERIES of proteins that
work in a cascade to assist in antibody destruction of
cells
The complement system is also part of the innate
immune system and can work as part of the
inflammatory system to directly eliminate a potential
pathogen.
- Deficiencies in each of the major complement
components have been described, leading to
various clinical symptoms
Deficiency of C2 - is believed to be the most common
complement component deficiency.
C3 deficiency - may also have a lupuslike clinical
presentation, but is more likely to involve recurrent
infections with encapsulated organisms.
A deficiency of C1 esterase inhibitor - has been found
in patients with hereditary neuroangioedema.
● Deficiencies of the later components of
complement (C5–C9) - are often associated
with recurrent Neisseria meningitidis
infections.
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Category 9: Phenocopies of Primary
Immunodeficiencies
This category comprises a new classification of PIDs.
Disorders that fall into this category have an inherited
genetic component but also include an acquired
component, such as:
➔ somatic mutations
➔ autoantibody production
Chronic mucocutaneous candidiasis, a disease that
was once classified as a cell-mediated deficiency, is
now included in Category 9.
- This disease is induced by a genetic mutation
in the AIRE gene, but also involves an
antibody to either (or both) IL-17 and IL-22.
● Disorders that fall into this category have an
inherited genetic component but also include
an acquired component, such as somatic
mutations or autoantibody production
● Chronic mucocutaneous candidiasis
●
Acquired Immunodeficiency syndrome (AIDS)
● Etiologic agent: Human immunodeficiency
virus (HIV) - The virus attacks the immune
system and leaves the body vulnerable to a
variety of life-threatening illnesses and prone
cancers
HIV - causative agent
AIDS - disease known as <-
BUT hindi porket meron HIV yung isang patients
meron na kaagad AIDS → nagkaroon lang ng AIDS if
CD4 T Cells count falls BELOW 200 (NORMAL → 1500
up to 3000) meaning hindi na kaya ng katawan,
immunocompromised na and have now AIDS, and the
patient are now VULNERABLE to different
LIFE-THREATENING ILLNESS
Human immunodeficiency virus (HIV) - is the etiologic
agent of acquired immunodeficiency syndrome, or
AIDS, a disease that has posed one of the greatest
medical challenges worldwide - because wala pang
cure and treatment is usually management
(pinapalakas ang immune system)
Transmission
➔ intimate sexual contact
➔ contact with blood or other body fluids, or
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➔ perinatally (from infected mother to infant)
- The majority of cases of HIV infection have
occurred through sexual transmission
involving either vaginal or anal intercourse
- transmission is perinatal, from infected
mother to her fetus or infant. Transmission
through this route can occur during
pregnancy, by transfer of blood at the time of
delivery, or through breastfeeding
The gag gene codes for p55, a precursor protein with
a molecular weight of 55 kd, from which four core
structural proteins are formed:
➔ p6
➔ p9
➔ p17
➔ p24
All four are located in the nucleocapsid of the virus.
The env gene codes - for the glycoproteins gp160,
gp120, and gp41, which are found in the viral
envelope.→ important for the ATTACHMENT of HIV to
CD4 cells

The third structural gene, pol, codes for enzymes

necessary for HIV replication

These include reverse transcriptase (p51);

ribonuclease
(RNase H; p66) - an enzyme involved in the
degradation of the original HIV RNA

integrase (p31) - an enzyme which mediates the

integration of viral DNA into the genome of infected
host cells

HIV belongs to the genus Lentivirinae of the virus protease (p10) - that cleaves precursor proteins into
family Retroviridae. smaller active units used to make the mature virions.
- It is classified as a retrovirus because it
contains ribonucleic acid (RNA) as its nucleic - These proteins are located in the core of the
acid and a unique enzyme, called reverse virus in association with HIV RNA.
transcriptase, which transcribes the viral RNA
into DNA, a necessary step in the virus’s life
cycle.
- HIV is a spherical particle, 100 to 120 nm in
diameter, which contains an inner core with
two copies of single-stranded RNA surrounded
by a protein coat or CAPSID and an outer
envelope of glycoproteins embedded in a lipid
bilayer
The genome of HIV includes three main structural
genes:
➔ gag,
➔ env
➔ pol
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T helper (Th) cells - are the MAIN TARGET for HIV

infection because they express high numbers of CD4
molecules on their surface and bind the virus with
high affinity

Other cells such as macrophages, monocytes,

dendritic cells, Langerhans cells, and microglial brain
cells can also be infected with HIV because they have
some surface CD4.

HIV viruses that preferentially infect T cells are known

as T-tropic or X4 strains, whereas those strains that
can infect both macrophages and T cells are called
M-tropic or R5 strains.

TROPIC/ TROPISM - means they only affect a SINGLE

TYPE of cell

Viral Replication → Entry of HIV into the host cells to which it has
attached requires an additional binding step, involving
co-receptors that promote fusion of the HIV envelope
with the plasma cell membrane.
- These co-receptors belong to a family of
proteins known as chemokine receptors,
whose main function is to direct white blood
cells (WBCs) to sites of inflammation.

CXCR4 - required for HIV to enter T lymphocytes

CCR5 - is required for entry into macrophages.

- In fact, individuals who have a genetic

The first step in the reproductive cycle of HIV occurs
when the virus attaches to a susceptible host cell.
This interaction is mediated through the host-cell CD4
antigen, which serves as a receptor for the virus by
binding the gp120 glycoprotein on the outer envelope
of HIV.
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mutationin the CCR5 gene have been found to
be resistant to HIV infection.
- Binding of the co-receptors allows for HIV
entry by inducing a conformational change in
the gp41 glycoprotein, which mediates fusion
of the virus to the cell membrane.
- After fusion occurs, the viral particle is taken
into the cell and uncoating of the particle
exposes the viral genome
- Action of the enzyme reverse transcriptase
produces complementary DNA from the viral
RNA.
- Double-stranded DNA is synthesized and, with
the help of the HIV integrase enzyme,
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becomes integrated into the host cell’s
genome as a provirus
- The provirus can remain in a latent state for a
long time, during which viral replication does
not occur. Eventually, expression of the viral
genes is induced when the infected host cell is
activated by binding to antigen or by exposure
to cytokines.
- Viral DNA within the cell nucleus is then
transcribed into genomic RNA and messenger
RNA (mRNA), which are transported to the
cytoplasm.
- Translation of mRNA occurs, with production
of viral precursor proteins and assembly of
viral particles.
- The intact virions bud out from the host cell
membrane and acquire their envelope during
the process.
- The precursor proteins are cleaved by the viral
protease enzyme in the mature virus particles.
- These viruses can proceed to infect additional
host cells.
- Viral replication occurs to the greatest extent
in antigen-activated Th cells. Because viral
replication occurs very rapidly and the reverse
transcriptase enzyme lacks proofreading
activity, genetic mutations occur at a high
rate, producing distinct isolates that exhibit an
extraordinary level of antigenic variation.
- In fact, the level of HIV diversity in a single
individual is greater than the diversity of all
the influenza virus isolates throughout the
world in a given year!
- This tremendous genetic diversity of HIV
hinders the ability of the host to mount an
effective immune response.
Immunologic Manifestations
● Initial viral replication: presence of increased
levels of p24 antigen
● As the virus replicates, some of the viral
proteins produced within host cells form
complexes with class I major
histocompatibility complex (MHC) antigens
and are transported to the cell surface, where
they stimulate lymphocyte responses.
● HIV-specific CD4+ Th cells are generated and
assist both humoral and cell-mediated
immune responses against the virus.
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B lymphocytes are stimulated to produce antibodies
to HIV, which can usually be detected in the host’s
serum by 6 weeks after primary infection.
● The first antibodies to be detected are
directed against the gp41 transmembrane
glycoprotein followed by production of
antibodies to the gag proteins such as p24,
and finally production of antibodies to the
env, pol, and regulatory proteins.
● T-cell–mediated immunity is thought to play
an important role in the immune response to
HIV
● CTL and antibody responses to HIV are
hindered by the virus’s ability to undergo
rapid genetic mutations, generating ESCAPE
mutants with altered antigens toward which
the host’s initial immune responses are
ineffective
● HIV Pro-viral state: HIV is protected from
attack by the immune system until cell
activation stimulates the virus to multiply and
display its viral antigens
- SILENCE PRO VIRUS for LONG PERIOD OF TIME
● The ability of HIV to evade the immune
response results in a persistent infection that
can destroy the immune system
SUMMARY
KEY TO IMMUNODEFICIENCIES
ADA: adenosine deaminase deficiency
AT: ataxia-telangiectasia
BTK: Bruton’s tyrosine kinase deficiency
CGD: chronic granulomatous disease
C-H: Chediak-Higashi syndrome
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CVI: common variable immunodeficiency

DiGeorge: DiGeorge anomaly
IgA def: selective IgA deficiency
IgG def: IgG subclass deficiency
JAK3: Janus kinase-3 deficiency
LAD: leukocyte adhesion deficiency
PNP: purine nucleoside phosphorylase deficiency
WAS: Wiskott-Aldrich syndrome
xSCID: X-linked severe combined
immunodeficiency disease
KEY TO CELLS
PS: pluripotent stem cell
CMP: common myeloid precursor
CLP: common lymphoid precursor
NK: natural killer cell
IB: immature B cell
IT: immature T cell
Th: helper T cell
Tc: cytotoxic T cell
PC: plasma cell

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AUTOIMMUNITY
OVERVIEW
● In the early 1900s - PAUL EHRLICH have noted
that our immune system could actually attack
the very own host or out very own self
→ which our immune system SHOULD
intended to protect BUT they are times that
our immune system becomes overwhelmed
due to different phenomena and mechanism
that’s why it could attack also the host or the
host tissue and during that time PAUL
EHRLICH referred that as fear or self poisoning
or → HORROR AUTOTOXICUS
● PAUL EHRLICH - the one who first described
the mechanism involved in autoimmunity or
in the different autoimmune diseases and
later on this conditions that causes damage or
attacks the very host occurred later and
became knowns as → AUTOIMMUNE DISEASE
● Self tolerance is term used to describe the
non-responsiveness of the immune system
when it encounters one’s own antigens.
(dapat hindi niya iaattack)
● AUTOIMMUNE DISEASE is a BREAKDOWN in
self tolerance can result in autoimmunity.
Autoimmune diseases
- DISORDERS in which immune responses are
targeted toward SELF- ANTIGENS and result in
damage to organs and tissues in the body.
SELF- ANTIGENS - host tissues or host cells, meaning
sarili mong tissues hindi nakikilala ang immune
systems, and immune systems treats that as FOREIGN
that's why it attacks self antigen causing damage to
organs and tissues wherein hindi naman dapat sinisira.
IMMUNE SYSTEMS should protect SELF ANTIGENS but
due to abnormalities in immune systems they were
not able to recognize the self antigen.
AUTO IMMUNITY - represents a breakdown in self
tolerance. represents a breakdown in immune
systems ability to discriminate self from nonself
antigen that’s why even self antigen were attacked
because immune system cannot recognize self from
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nonself and usually these harmful effects because of
that mechanism and conditions can be caused by
→ T cell mediated immune responses as well as →
humoral mediated immune responses in the form of
AUTOANTIBODIES that are directed against HOST
ANTIGENS or SELF ANTIGENS so in T CELL MEDIATED,
t cells → attack the cells they secrete CYTOKINES
attacking SELF ANTIGEN as well as the production of
autoantibodies directed against host or self antigens.
During the differentiation of → T CELLS and B CELLS
there could actually be ABNORMALITIES during the
positive and negative selection → during these
mechanism pinapa-kilalala sa T CELLS yung ating self
antigens so that pag labas na ng THYMUS there will
able recognize their own antigen → BUT if may
abnormalities during positive and negative it could be
a REASON to acquired or to cause an autoimmune
disease whether t cell mediated or antibody mediated
response against our host antigens so basically that is
→ AUTOIMMUNE DISEASES or AUTOIMMUNITY.
Autoimmunity
●
● Conditions in which DAMAGE to → organs
and or tissues results from the presence of \
→ autoantibody or autoreactive cells.
AUTOANTIBODY - these are antibodies that are
directed AGAINST our self antigen or host antigens.
(meaning dapat hindi siya dapat andoon, wala dapat
tayong antibody against self antigen but there are
time wherein nag produce ng katawan
AUTOANTIBODIES which are antibodies directed to
our self antigen and that causes damage to the host
tissue causing → AUTOIMMUNE DISORDER
● Most autoimmune diseases exhibit MARKED
familial incidence suggesting → genetic
predisposition
●
● Autoimmune responses are often related to
the presence of specific → major
histocompatibility complex (MHC)
eg: if you inherit a certain GENE which is link to an
autoimmune disorder you have INCREASED risk of
developing that autoimmune disorder since there are
studies that autoimmune diseases have → FAMILIAL
INCIDENTS of these specific MAJOR
HISTOCOMPATIBILITY COMPLEXES (MHC)
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accepts self antigen and do NOT initiate a

response against them)

Etiology - DEFECT in mechanisms underlying self

recognition
→ cause of AUTOIMMUNE DISEASE, hindi nakikilala ng
sarili mong katawan/immune system yung SELF
ANTIGEN and it damage and causing LOCAL DAMAGE
or causing diseases in the body and usually → 5-7%
ADULTS are AFFECTED with autoimmune diseases and
usually WOMEN are more affected than MEN.
In producing or acquisition of autoimmune disorder
and we have a lot or → more than 40 human diseases
of autoimmune origin
SLE and CRA → rheumatoid arthritis are autoimmune
diseases which are also a part of HYPERSENSITIVITY
REACTION but the origin, is that yung autoantibody at
yung self antigen nag cocombine ng mag deposit sa
tissues that’s why it also a TYPE 3 HYPERSENSITIVITY
REACTION but the origin of the disease is an
→ AUTOIMMUNE origin
ETIOLOGY OF AUTOIMMUNE DISEASE
●
● The main etiology of autoimmune disorder is
the BREAKDOWN of → SELF-RECOGNITION
and SELF-TOLERANCE
●
Self-Tolerance
Generalize reason why we produce or why we
develop autoimmune disorders
SELF TOLERANCE - ability of the immune system to
NOT respond or a NON-RESPONSIVE mechanism of
the immune system to self antigen.
● Autoimmunity can be defined as BREAKDOWN
of mechanisms responsible for SELF
TOLERANCE and induction of an immune
response against components of the self.
●
● Self-tolerance is a type of → immunologic
tolerance, or a state of immune
UNRESPONSIVENESS that is directed against a
specific antigen or a self-antigen (na dapat
hindi mag rereact ang immune system to
self-antigen) (Immune system SHOULD
LYMPHOCYTES - must be educated so they can
distinguish between self and non self antigen.
These programming of LYMPHOCYTES to distinguish or
differentiate between SELF ANTIGEN and FOREIGN
ANTIGEN can take place under TWO LEVELS ↓
Two levels:
○ Central tolerance - T Cells become programmed to
differentiate between SELF from NONSELF in the →
PRIMARY LYMPHOID ORGAN
○ Peripheral tolerance - this happen on →
SECONDARY LYMPHOID ORGANS and that happen in
SPLEEN and LYMPH NODES
There are also autoimmune disorder that are
→ IDIOTYPIC - meaning they may NOT always be
harmful but numerous diseases are well recognized to
produce damage, so hindi naman lahat ng
autoimmune disorder causes damage but MAJORITY
of the autoimmune diseases causes systemic damages
to our body.
Central Tolerance
● It occurs in the CENTRAL or PRIMARY
LYMPHOID ORGAN : the → THYMUS (T Cells),
and the BONE MARROW (B Cells)
→ NOT all T cells are fully functional and are fully able
to eliminate self reactive cells during the negative
selection that’s why doon nagkakaroon ESCAPE of
auto reactive clones which can result now to →
autoimmune disorder
NOT all self antigens may be represented in →
THYMUS or there are certain antigens that may NOT
be properly process and presented during negative
selection → that’s why nagkakaroon ng problema
during the T cells mediated activity pag labas sa
katawan because of the problem encountered during
the → CENTRAL TOLERANCE

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All T cells that damages/reactive to self are DELETED
by → APOPTOSIS
but NOT all T-cells are deleted so some of the
self-reactive T cells instead of being deleted through
apoptosis they instead DIFFERENTIATE into → T
regulatory cells can specifically inhibit an immune
response to self antigen
happen in the → BONE MARROW in B cells as B cells
mature those B cells with the receptors having a
STRONG AFFINITY for self antigens are also eliminated
by APOPTOSIS during differentiation from
→ immature to mature b cells BUT as well NOT all B
cells are not cell reactive B Cells are deleted but
instead they also stimulate to REARRANGE their
receptor genes or their IMMUNOGLOBULIN GENES so
their B cells receptor are NO longer antigen specific
and that is known → RECEPTOR EDITING so that B
cells receptors NO longer act to our self antigens
Anergy
- DOWN REGULATION of B cell expression and
UNRESPONSIVENESS to the antigens
- so yung mga B cells na nag possess ng
receptors that only weakly recognize self
antigen and down regulate their expression on
receptor and develop a specific
UNRESPONSIVENESS to antigens are known
as → ANERGY
Central Tolerance - this process are NOT totally
functional at all times and NOT totally effective at all
times
- there are some self-reactive lymphocytes that
are able to manage to ESCAPE of auto reactive
clones from primary lymphoid organs to
secondary lymphoid organs such as spleen
and lymph nodes
Peripheral Tolerance
SECOND LEVEL of protection, happens in →
SECONDARY LYMPHOID ORGANS
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Peripheral Tolerance - SECOND LEVEL of protection
wherein LYMPHOCYTES that recognize self antigens in
the SECONDARY LYMPHOID ORGANS are rendered
incapable of reacting with self antigens.
PERIPHERAL TOLERANCE of T cells → can result from
ALLERGIES or nado-down regulate yung kanilang
expression and they produce a state of
UNRESPONSIVENESS or ANERGY cause → by the
ABSENCE CO-STIMULATORY SIGNAL from an antigen
presenting cells
→ since antigen presenting cells they are able to
recognize our self antigen so hindi sila mag bibigay ng
signal or inhibit nila yung mga receptors nila on the T
cells so that is mechanism involved in
→ Peripheral Tolerance
Peripheral T cells tolerance → can also result from
inhibition by T REGULATORY CELLS or they can be
programmed to → CELL DEATH
It could be ABSENCE CO-STIMULATORY SIGNAL on
Peripheral from antigen presenting cells as well as
INHIBITION through the → T regulatory cells or
DEATH BY APOPTOSIS
● Second level of tolerance occurring in the
secondary lymphoid organs
SELF TOLERANCE
SELF TOLERANCE → these are the state of
UNRESPONSIVENESS to self antigen.
→ so self-tolerance can be achieved CENTRALLY or
PERIPHERAL TOLERANCE
when we say CENTRALLY → it is happens on the
THYMUS or in the primary lymphoid organs for
T cell → thymus
B cells→ bone marrow
→ and then central tolerance may NOT be fully
functional so meron pa tayong → second level of
tolerance which happens on the → secondary
lymphoid organ the SPLEEN and LYMPH NODES and
that is known as a → PERIPHERAL TOLERANCE
→ BUT in some individuals BOTH SELF TOLERANCE on
the FIRST LEVEL and the SECOND LEVEL → so both
self-tolerance can FAIL even after the → second layer
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of protection or the peripheral tolerance so pag nag
fail parin or nag escape and the second layer of
tolerance dito na nagkaroon ng → AUTOIMMUNE
DISEASE or talaga magkakaroon na ng SELF MEDIATE
RESPONSE and HUMORAL MEDIATED RESPONSE ang
ating katawan against → SELF ANTIGEN
POSITIVE SELECTION - they should be able to
recognize in relation or association to MHC and TCR
should be functional
→ And after a positive selection meron pa ulit SECOND
PURGING OF CLONES which is the
→ NEGATIVE SELECTION - wherein all T cell that react
to self peptide in a damaging way will be DELETED
During the → NEGATIVE SELECTION all T cells that are
autoreactive T cells will be DELETED through
APOPTOSIS but NOT all T cells were DELETED some of
those autoreactive t cells becomes or differentiate
into t-regulatory cells wherein they also produced
→ inhibition of reacting to self antigen
● In some individuals, self-tolerance can fail
even after this second layer of protection;
● If this happens, AUTOIMMUNITY can arise
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Genetic Predisposition for Autoimmunity:
● Association between certain HLA types and
autoimmune diseases has been noted.
●
● Major histocompatibility complex (MHC)
products also seem to → influence antigen
recognition or nonrecognition by determining
the type of peptides that can be presented to
the T cells
AUTOIMMUNE DISEASES - are often more prevalent
in a FAMILY MEMBERS than among unrelated
individuals and are of course → more prevalent
among monozygotic TWINS than in non identical twins
or siblings
→ Very strong ang relation that HLA or the Type of
HLA antigens that inherit makes you → at risk or
increases your RISK of developing a particular
autoimmune disorder
eg: HLA b27 and ANKYLOSING SPONDYLITIS which is
an auto inflammatory disease that affects SPINE
HLA DR2 → which predisposes to MULTIPLE
SCLEROSIS (MS) → potentially disabling disease of
the brain and spinal cord → and SLE
There are also studies linking HLA DR4 and RA with
type 1 DIABETES MELLITUS and RHEUMATOID
ARTHRITIS → so there are different HLA types or
inherited HLA GENES that predisposes to certain →
autoimmune disorders
HLA DR4 → which predisposes to HASHIMOTO
THYROIDITIS
inheritance of these SPECIFIC GENES may make you
VULNERABLE or may make you more SUSCEPTIBLE
increases ONLY A RISK for → acquiring particular
autoimmune disease.
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● Expression of → class II molecules on cells stimulate → a both HUMORAL and CELL MEDIATED
where they are NOT normally found may IMMUNE RESPONSE
result in the presentation of self-antigens for
● The stimulatory effects of FEMALE
which NO tolerance has been established
HORMONES may place women at a GREATER
●
RISK for developing autoimmune disease
● Therefore, inheritance of a gene coding for a
→ specific MHC molecule may make an
individual MORE susceptible to a particular
autoimmune disease

Other Endogenous and Environmental Factors for

Autoimmunity
➔ Hormonal influence
➔ Tissue Trauma and Release of Cryptic
Antigens
➔ Microbial Infections
➔ Epigenetics
➔ Modification of Self-Antigens
But generally the cause etiology of autoimmunity is
→ BREAKDOWN of SELF TOLERANCE but there are also
factors that contributes → to INCREASE RISK of
developing an autoimmune disorder
Hormonal Influence
● HORMONES can influence also development
of autoimmune diseases → Women are 2.7
times more likely to acquire an autoimmune
disease than men
● About 78% of patients with autoimmune
diseases are of → FEMALE GENDER
WOMEN also tend to develop autoimmunity at an
EARLIER AGE and have a HIGHER RISK → for acquiring
more than one autoimmune disease as compared
with men based on previous or past studies
There is an EFFECT OF HORMONES like the ESTROGEN
in FEMALE that tends to → direct our immune system
in favor of a T CELLS the response resulting in a more
stimulation of our B-CELL ACTIVATION and more
antibody production
whereas ANDROGENS favor a type 1 helper responds
with activation of T cells
PROLACTIN → which is secreted by FEMALE is a
hormone that stimulate a production of BREASTMILK
in pregnant and nursing women and also can
●
Sequestered antigen or Release of Cryptic Antigens:
CRYPTIC - hidden within the tissue of the host
→ So during the development of LYMPHOCYTES in
the THYMUS and BONE MARROW when immunologic
tolerance or central tolerance occurs these
lymphocytes → Lymphoid cells may NOT be exposed
to some self antigen during their differentiation,
because these SELF ANTIGENS are HIDDEN WITHIN
the TISSUES and they may be late-developing antigens
or may be confined to specialized organs.
eg: yung mga self antigens on the EYES which are
sequestered or which are HIDDEN at napupunta lang
sa circulation if there is or following an ocular INJURY
or those that are from the the MYELIN BASIC PROTEIN
which are can only be found on CENTRAL NERVOUS
SYSTEM and normally sequestered by the
BLOOD-BRAIN BARRIER
→ but in cases of TRAUMA to these tissues by
→ accident or infection they may be introduced to the
GENERAL CIRCULATION and because these
self-antigen are NOT exposed to some of our
lymphocytes during their differentiation in the central
tolerance they are not able to recognize these HIDDEN
or CRYPTIC antigen so that as foreign antigen and they
initiate now an autoimmune disease so that could be
a factor
SPERM → during VASECTOMY can also produce auto
antibodies to SPERM
DNA → following damage to SKIN cells since our skin
are NOT exposed to the general circulation or
overexposure to UV RAYS from the sun can also

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produce auto antibodies to DNA following damage to STREP. PYOGENES one of it's complication or SEVERE
our SKIN complication are RHEUMATIC FEVER

● A release of antigens from these organs RHEUMATIC FEVER - is an autoimmune disorder as

resulting from accidental traumatic injury or well that primary affects the JOINTS and HEART
surgery can result in the stimulation of an
SCARLET FEVER or PHARYNGITIS → as a result of
immune response and initiation of an
STREPTOCOCCUS PYOGENES will proceed to develop
autoimmune disease.
→ RHEUMATIC FEVER if they are NOT treated
●
adequately by ANTIBIOTICS
● This concept has also been referred to as
→ immunologic ignorance and may be SYMPTOMS RHEUMATIC FEVER are thought to be
responsible for the production of cause by the → production of antibodies to
autoantibodies or production of ANTIBODIES
against SELF ANTIGENS
●

Microbial Infections
Another factor that may contribute the development
of an autoimmune disorder is
→ INFECTION/MICROBIAL INFECTION
Molecular Mimicry
● Refers to the fact that → MANY BACTERIAL or
VIRAL AGENTS contain antigens that closely
resemble the structure or amino acid
sequence of self-antigen. (KAHAWIG- that’s
NALILITO ngayon ang immune system
napagkamalan nila na yung self antigen natin
na SIMILAR ang structure to the particular
bacteria na naexpose sila previously so may
memory, so dahil same ng structure they will
regard self antigen na BACTERIAL ANTIGEN
that’s why they will attack it
→ MOLECULAR MIMICRY
●
● Exposure to such → FOREIGN ANTIGENS may
TRIGGER immune responses that
CROSS-REACT with similar self antigens.
Mechanism involved → CROSS-REACTION
MECHANISM wherein our antibody to certain bacteria
and viral agents can also to our self antigen for which
they have RESEMBLANCE or they have → SIMILAR
amino acid sequence and structure
eg: association of our gram-positive STREPTOCOCCUS
PYOGENES and RHEUMATIC FEVER
→ M PROTEIN and N-ACETYL GLUCOSAMINE
components of the bacteria which is cross-reacting to
→ the CARDIAC MYOSIN that's why akala ng ating
immune system that our CARDIAC MYOSIN is the same
as the M PROTEIN and N-ACETYL GLUCOSAMINE of
strep. pyogenes inaattack niya din ang → CARDIAC
MYOSIN causing damage to the → HEART
so the ORIGIN of rheumatic fever is because S.
PYOGENES regards the CARDIAC MYOSIN of the
HEART as the M PROTEIN and N-ACETYL
GLUCOSAMINE of the bacteria
CROSS REACTION MECHANISM of our SELF ANTIGEN
with other microbial agents
eg: POLIOVIRUS which also resembles the
ACETYLCHOLINE RECEPTORS as well as → MEASLES
VIRUS which can CROSS REACT to the → MYELIN
BASIC PROTEIN that's our self antigen that's why akala
ng katawan myelin basic protein are MEASLES VIRUS
that's why it ATTACKS
PAPILLOMAVIRUS - which predisposes or INCREASES
RISK to produce → DIABETES MELLITUS because
INSULIN RECEPTOR can cross-react the
PAPILLOMAVIRUS AGENTS
BYSTANDER EFFECT

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→ a SECOND WAY that MICROBES might TRIGGER an
autoimmunity is through → BYSTANDER EFFECT
● The microbial organism do NOT share
structurally similar antigens with the host.
●
● Instead, the microorganism can induce a
LOCAL INFLAMMATORY RESPONSE that
recruits → leukocytes and stimulates APCs
(ANTIGEN PRESENTING CELLS) to release
cytokines that nonspecifically → activate T
cells
T CELLS ACTIVATED → May have specificity for self
antigens and these now will react the self antigens
causing → damage to TISSUES and these activation of
immune response to UNRELATED antigens has also
been termed as EPITOPE SPREADING or BYSTANDER
EFFECT
● The agent do NOT necessarily need to share a
→ similar antigen with the host or hindi
naman cross-reacting but the inflammatory
response is through the INDUCTION or
recruitment of leukocytes and stimulation of
antigen-presenting cell to release →
CYTOKINES
● Non-specifically activate T cells → hindi dahil
may antigen na pinepresent BUT they
nonspecifically activate the t cells and these T
cells that are activated may have specificity for
self antigens causing → damage to that host
tissue or host cells so that is → BYSTANDER
EFFECT or EPITOPE SPREADING
(Non-specifically activate T cells by these
MICROBIAL ORGANISM na hindi naman dapat)
SUPERANTIGENS
a THIRD WAY that MICROORGANISMS might INDUCE
an autoimmunity is through formation of
→ SUPERANTIGENS
● These are PROTEINS that are produced by
various microbes that have the ability to bind
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99
to → BOTH class II MHC molecules and TCRs,
regardless of their antigen specificity.
eg: STAPHYLOCOCCAL ENTEROTOXIN → that causes
FOOD POISONING and TOXIC SHOCK SYNDROME
These superantigen can act as a POTENT T-cells
MITOGENS → by activating a LARGE NUMBER of T
cells with different antigens specificities
So, if some of these t cells POSSESS specificity for a
→ self-antigen at an autoimmune response might
result
There are able to BIND to both CLASS ll MHC
MOLECULES and → activate T cells and activate a
large number of t cells with different antigen
specificity → if these T cells possess specificity for a
certain self-antigen an autoimmune response may be
INDUCED
POLYCLONAL B CELL ACTIVATION
→ such as EPSTEIN BARR VIRUS and
CYTOMEGALOVIRUS → they can cause POLYCLONAL
ACTIVATION OF B-CELLS and INDUCE proliferation of
numerous clones of b cells that express IgM in the
→ absence of the T helper
And in this process the B-CELLS may be reactive to
self antigens so B-cells can be activated when will be
reactive to self antigens
● B-cell defects include the ABNORMAL
EXPRESSION or function of KEY signaling
molecules, dysregulation of cytokines, and
changes in B-cell developmental subsets.
●
● FC receptor polymorphisms, the receptor for
antibody that normally down-regulates
antibody production, may cause → continual
B-cell stimulation.
●
● These defects may be ENCHANCED by
organisms such as gram-negative bacteria and
several viruses, including cytomegalovirus
 IMMUNOLOGY AND SEROLOGY
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and EpsteinBarr virus (EBV), which are ability to be processed by → APCs and
→ polyclonal activators presented to T cells

Epigenetics and Modification of Self-Antigens AUTOIMMUNITY

The etiology of autoimmunity is UNKNOWN

● Epigenetics → refers to modifications in GENE → There are several mechanism that are involved
EXPRESSION that are NOT caused by changes production of an autoimmune disorder
in the original DNA sequence so these
→ it's basically a disease entity which is caused by a
alteration are STABLE and can be inherited
COMPLEX INTERACTION between GENETIC and
●
ENVIRONMENTAL INTERACTION as well as immune
● Over or under expression of certain genes in
regulation so there are certain genes make an
the immune system may result in →
individual more susceptible to immune response
homeostatic imbalances and a breakdown of
against → self-antigen but are NOT sufficient by
self-tolerance, leading to → autoimmunity, so
themselves that's why environment GENDER of
basically inherited GENE EXPRESSION can also
individual tissue injury exposure to infectious agents
TRIGGERED formation of an autoimmune
other environmental agents have also been significant
response
and can TRIGGER an immune response in a
susceptible individual

BREAKDOWN in → immunologic tolerance escape of

other reactive cells and also abnormal recognition and
proliferation in responses to self antigens of our b cells
as well as our t cells can also perpetuate the
autoimmune response nagkakaroon ng dysfunction
●
● Post-translational modifications → refers to → if magkaroon ng DYSFUNCTION in regulation of
changes at the PROTEIN LEVEL which can be cells or it will also result to an → autoimmune
sometimes due to exposure to environmental response
factors.
The development of autoimmune response or disease
The changes that are involved in Post-translational is thought to be a → CAUSE of COMPLEX INTERACTION
modifications may involve → biochemical processes between GENETICS EXPOSURE to → different
such as: ENVIRONMENTAL FACTOR and dysfunction and effects
in our immune regulation
➔ ACETYLATION
➔ LIPIDATION COMPLEX INTERACTION:
➔ GLYCOSYLATION → that’s why nagkakaroon
➔ ENVIRONMENTAL FACTOR
ng modification which alters its ➔ GENETICS EXPOSURE
IMMUNOGENICITY of our self antigen ➔ DEFECTS OF IMMUNE REGULATION
affecting its ability to be processed by
→ APCs and that is why our body REGARDS
them as NON-SELF ANTIGENS → leading
autoimmune response
➔
● This can lead to alteration of the
immunogenicity of an antigen, affecting its

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• Clinical and serological • OVERLAP of SLE, RA and

overlap (e.g. Thyroid, other connective tissue
stomach, adrenal glands, disorders
kidney) → ORGAN
SPECIFIC • Antigens are accessible
at HIGHER
• Antigens are ONLY CONCENTRATION
available to lymphoid
system in low • No antibodies produced
concentration in animals with
comparable stimulation
General classification:
• Antigens evoke
Autoimmune diseases are generally classified on the • Familial tendency to
organ-specific antibodies
basis of the organ or tissue involved. develop connective tissue
in normal animals with
These disease may fall in: complete Freund’s
adjuvant • Questionable
ORGAN-SPECIFIC → category in which the immune abnormalities in Ig
response is DIRECTED against antigen(s) associated • Familial tendency to synthesis
→ with the one TARGET ORGANS being damaged develop organ-specific
autoimmunity • Lesions caused by →
NON ORGAN-SPECIFIC/ SYSTEMIC → category in antigen-antibody
which the antibody is directed against an antigen NOT • Lymphoid invasion, complexes
associated with the target organ → ALL OVER THE parenchymal destruction
by questionable • Tendency to develop
BODY or MULTIPLE ORGAN
cell-mediated lymphoreticular
hypersensitivity or neoplasia
antibodies

• Tendency to develop
→ cancer in organ

Organ Specific Autoimmune Diseases

Organ specific
These AUTOIMMUNE DISEASES have a → PARTICULAR
TARGET ORGAN wherein the autoantibodies are
DIRECTED

AUTOIMMUNE DISEASES

Organ-Specific Systemic / Non-Organ

Specific

• Antibodies and lesions • Antibodies and lesions

are directed towards a → are NOT CONFINED to
SINGLE TARGET ORGAN any organ

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Addison’s disease
Addison’s disease - low secretion of SODIUM →
magkakaroon pag may HYPONATREMIA →
DEFICIENCY in ALDOSTERONE and SODIUM
Addison’s disease TARGETS → Adrenal glands
(secrete ALDOSTERONE) → so nag produce tayo ng
→ ANTIBODY against the ADRENAL CELLS
Celiac disease
Celiac disease - autoimmune disorder where
→ Small intestine and other organs are AFFECTED
Celiac disease - is UNIQUE and associated with an
ENVIRONMENTAL TRIGGER → TRIGGER of Celiac
disease is GLUTEN or DIETARY GLUTEN
GLUTEN - is a protein complex found in WHEAT that is
POORLY DIGESTED by → UPPER GASTROINTESTINAL
TRACT → and GLUTEN contains SOLUBLE COMPONENT
that is → GLIADIN that is RICH in AMINO ACID
GLUTAMINE and PROLINE
GLIADIN - is RESISTANT to digestive enzymes in
STOMACH and SMALL INTESTINE after ingestion
If there is an increase in permeability of the intestinal
wall → nagkakaroon ng result an INFECTION because
of the → UNDIGESTED GLIADIN
that's why parang sa nagiging sequestered antigen the
immunogenicity of gliadin is ENHANCED when it is
acted by the tissue → TRANSGLUTAMINASE because
of this nagkakaroon ng generation specifically HLA
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102
DQ2 and HLA DQ8 on antigen-presenting cell
necessary for developing → CELIAC DISEASE
CELIAC DISEASE produce:
➔ Anti Transglutaminase (tTG)
➔ Antibodies to deamidated gliadin peptides
(DGPs)
➔ Endomysial antibodies
GOODPASTURE’s SYNDROME
Goodpasture’s syndrome - autoimmune disease as
well that have the presence of an autoantibody to
→ GLOMERULAR RENAL TUBULAR and ALVEOLAR
BASEMENT MEMBRANE → resulting to PRIMARY
INJURY to GLOMERULUS that can rapidly progress to
RENAL FAILURE and also can → infect or affect the
ALVEOLAR BASEMENT MEMBRANE of the LUNGS
➔ GLOMERULAR RENAL TUBULAR
➔ ALVEOLAR BASEMENT MEMBRANE of the
LUNGS and KIDNEYS
GRAVES DISEASE
Graves disease - is also an autoantibody or it's an
autoimmune disorder wherein meron
HYPERTHYROIDISM → state of excessive THYROID
HORMONES because there is an excessive thyroid
function
THYROTOXICOSIS → there is an excess amount of
thyroid hormones with a diffusive ENLARGE GOITER
that is firm instead of RUBBERY
35% of patients with GRAVES DISEASE is
EXOPHTHALMOS in which there is HYPERTROPHY of
the EYES or the EYE MUSCLES and increased
connective tissue in the orbit causing the → EYEBALL
to BULGE OUT so that the patient has LARGE EYE
staring expression
MAJOR AUTOANTIBODY INVOLVED ARE:
➔ Thyroid-stimulating hormone receptor
antibodies (TRAbs)
➔ Antithyroglobulin
➔ Antithyroid peroxidase (TPO)
 IMMUNOLOGY AND SEROLOGY
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THYROID HORMONES → it is stimulated by the

THYROGLOBULIN & THYROID PEROXIDASE → are
needed in production of our thyroid hormones
HASHIMOTO'S THYROIDITIS
Hashimoto’s thyroiditis - immune destruction of the
thyroid gland occurs also known as → LYMPHOCYTIC
THYROIDITIS which results in a state of DECREASE
thyroid function and called as HYPOTHYROIDISM
They also have a goiter or enlargement of the thyroid
gland but the difference between the two is the
AMOUNT of thyroid hormones that they are
producing
SYMPTOMS OF HASHIMOTO'S THYROIDITIS:
➔ DRY SKIN
➔ PUFFY FACE
➔ EDEMATOUS OF EYELIDS
➔ POLAR
➔ WEIGHT GAIN
➔ FATIGUE
➔ DRY AND BRITTLE HAIR
HYPOTHALAMUS in the BRAIN or in the CENTRAL
NERVOUS SYSTEM and then the hormones that are
produced by the HYPOTHALAMUS will stimulate the
PITUITARY GLAND to → secrete the thyroid
stimulating hormone.
and the THYROID STIMULATING HORMONE → have
receptors for the thyroid, so sa THYROID GLAND
meron receptor ang TSH once mag bind si TSH your
thyroid gland → will now stimulate to produce
→ thyroid hormones in the form of
→ triiodothyronine and tetraiodothyronine
(T3 and T4) and to be able to → produce thyroid
hormones it needs an enzymes that what we call
→ THYROID PEROXIDASE and THYROGLOBULIN so
that IODINE and THYROGLOBULIN will → FUSED
GRAVES DISEASE → Thyroid stimulating hormone
receptor antibodies MIMICS the action of TSH
→ That's why they will stimulate, UNCONTROLLED
STIMULATION of thyroid hormones or release
excessive thyroid hormones → na OVERSTIMULATE
kaya nasisira siya

Organ specific

GRAVES DISEASE - Mechanism involves the

production of an autoantibody that what we called
→ THYROID STIMULATING HORMONE RECEPTOR
ANTIBODIES

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MULTIPLE SCLEROSIS
Multiple sclerosis - Is a autoimmune disorder
involving inflammation and destruction of the
→ CENTRAL NERVOUS SYSTEM
→ it is characterized by formation of LESIONS that
what we call → PLAQUES in the white matter of the
BRAINS and SPINAL CORD resulting in damage and
destruction of MYELIN SHEATH of the NERVES because
of the production of antibodies to the → myelin basic
protein
→ So the antibody binds to the → myelin membrane
and initiate an → immune response stimulating
phagocytes and other macrophages what we call
MICROGLIAL CELLS to produce immunologic events
which will result now to ACUTE INFLAMMATION
within the → AXONS and INJURY to the GANGLIA, so
nagkakaroon ng DYSFUNCTION and
POST-INFLAMMATORY NEURODEGENERATION so
affected ang CNS with MULTIPLE SCLEROSIS
T helper 1 cytokines - produced by the T helper 1 such
as:
➔ INTERLEUKIN 1
➔ TUMOR NECROTIC FACTOR ALPHA
➔ INTERLEUKIN GAMMA OR BELIEF to be central
pathogenesis of your multiple
→ also ENVIRONMENTAL FACTOR have been noted
such as → reduce exposure to sunlight → VITAMIN D
DEFICIENCY → can also predispose to a development
of MULTIPLE SCLEROSIS (MS)
➔ VISUAL DISTURBANCES
➔ DIMINISH LOCOMOTOR COORDINATION
LIMBS
➔ DIZZINESS
➔ FACIAL PALSY
➔ TINGLING - unusual sensation most commonly
felt in your hands, feet, arms and legs
➔ EXTREMITIES/PINS and NEEDLES that run
down to the SPINE
➔ FLASHES OF LIGHTS seen on EYE MOVEMENT
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MYASTHENIA GRAVIS
Myasthenia gravis - Muscles involved
➔ MUSCLE WEAKNESS IN THE UPPER LIMBS
➔ DIFFICULTY IN SPEAKING, CHEWING,
SWALLOWING
➔ UNABLE TO MAINTAIN SUPPORT OF TRUNK,
NECK & HEAD
➔ RESPIRATORY MUSCLES WEAKNESS (life
threatening progression)
➔ POLIO VIRUS
PERNICIOUS ANEMIA
Pernicious anemia - there is a destruction of the
parietal cells of the stomach leading to → intrinsic
factor deficiency
POSTSTREPTOCOCCAL GLOMERULONEPHRITIS
Poststreptococcal glomerulonephritis - Streptococcal
antibodies that cross-react with kidney tissue
→ MOLECULAR MIMIC of strep antigens that
cross-react with a tissue of the kidney that's why the
KIDNEY are damage
Type 1 diabetes mellitus
Type 1 diabetes mellitus - AUTOIMMUNE, affecting
pancreas , kaya mataas ang GLUCOSE is because hindi
nag produce ang PANCREAS ng INSULIN
INSULIN - needed for the glucose to be taken up by
the cell and to be used up by ENERGY
→ but if there is NO INSULIN in the body → your
glucose remain in the blood that's why constantly your
glucose ELEVATED in the blood causing →
HYPERGLYCEMIA
Type 2 diabetes mellitus - INSULIN RESISTANCE
because of the sedentary LIFESTYLE nasisira yung
PANCREAS (MAJORITY) - nag poproduce pero konti
lang
 IMMUNOLOGY AND SEROLOGY
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Systemic Autoimmune Diseases
Systemic Lupus Erythematosus (SLE) - there is
antibodies to → DNA and other nuclear components
such as phospholipids
Osteoarthritis → because of the GENERATION or
nasisira yung CARTILAGE → GENERATIVE DISORDER
Rheumatoid Arthritis → kahit young age pwede
magkaroon INFLAMMATION sa JOINTS → cause
DEPOSITION OF IMMUNE COMPLEX → FORMATION
OF AUTOIMMUNE DISEASE
Systemic or Non-organ Specific
Rheumatoid Arthritis
● Chronic, inflammatory joint disease with
systemic involvement.
RA → is another example of a systemic autoimmune
disorder. It affects about 0.5% to 1.0% of the adult
population, but prevalence varies with ethnicity and
geographic location.
→ It strikes individuals between the ages of 25 and 55.
→ WOMEN are 3x (three times) as likely to be affected
as men;
In addition, the prevalence of the disease is highest in
WOMEN who are more than 65 years of age.
● destruction of bone and cartilage.
● A symmetric, and erosive arthritis of the
PERIPHERAL JOINTS that can also affect
→ multiple organs such as the HEART and the
LUNGS
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● It's hallmark → feature is the presence of
→ rheumatoid factors, are antibody against
IgG immunoglobulin that is produced by
→ B cells and plasma cells in the synovial
membrane.
○ Mixture of organ specific with systemic
symptoms
○ Inflamed, swollen and painful joints
○ RF – IgM to the fc region of IgG
→ It is not known what role autoantibodies play in the
initiation of the inflammatory response.
→ Two key antibodies found in the disease are RF and
anti-CCP.
→ RF is an antibody that is most often of the IgM class
and is directed AGAINST the FC portion of IgG.
→ It has been postulated that RFs may play a role in
the pathogenesis of RA by → increasing macrophage
activity and enhancing antigen presentation to T cells
by APCs.
ETIOLOGY OF RA
● The STRONGEST GENETIC associations have
been between a subset of patients with RA
and specific HLA-DRB1 alleles or PTPN22 gene
polymorphisms.
●
● The strongest environmental risk factor for RA
is believed to be CIGARETTE SMOKING, which
doubles the risk of developing the disease.
Clinical Significance:
● The pathology of RA is caused by an
inflammatory process that results in the
●
● Two key antibodies found in the disease are
RF and anti-CCP.
 IMMUNOLOGY AND SEROLOGY
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○ RF → is an antibody that is most often of the
IgM class and is directed against the Fc
portion of IgG
→ This immune complex becomes deposited → to
the joints and now a sheet of inflammatory granulated
tissues grows → into the joint space and invades the
cartilage causing → the immunologic activity within
the host and increased inflammation due → to the
continual activation of certain immune cells secreting
→ pro-inflammatory cytokines secreting different
inflammatory processes stimulating production of
cytokines facilitating recruitment and transport →
wbc to the affected areas that's why nagkakaroon ng
→ local damage and inflammation within the joints
→ and that’s HYPERSENSITIVITY TYPE 3 but the origin
nagkakaroon ng DEPOSITION in the JOINTS is an
AUTOIMMUNE ORIGIN production of →
RHEUMATOID FACTOR
○ Antibodies to cyclic citrullinated proteins
(anticyclic citrullinated peptide antibody
[anti-CCP or ACPA]) → These antibodies can
react with citrulline-containing components of
the matrix, including filaggrin, keratin,
fibrinogen, and vimentin, and are thought to
correlate with the pathogenesis of RA
Treatment
● The discovery that joint destruction occurs
early in the disease has prompted more
aggressive treatment and the development of
new drugs to prevent disease progression.
→ In the past, therapy for RA was primarily based on
nonsteroidal anti-inflammatory drugs (NSAIDs) such
as salicylates (aspirin) and ibuprofen. Although these
agents can be used to → reduce local swelling and
pain, they are → NO longer the dominant treatment
for RA.
● Disease-modifying anti-rheumatic drugs
(DMARDs), most notably methotrexate, are
now prescribed at the time of diagnosis.
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→ If methotrexate alone does NOT work, it can be
combined with other → DMARDs or with biological
agents.
DMARDs and biological agents have → become the
mainstay of RA treatment because → They act on
specific components of the inflammatory response
(MACROPHAGE, T-CELLS, B-CELL) and are effective in
slowing the progression of joint erosion → because of
IMMUNE RESPONSIVENESS or
HYPERSENSITIVITY/EXAGGERATED IMMUNE
RESPONSE
NO CURE FOR RA, → just manage progression and
reduce the inflammation
● Methotrexate is thought to act by inhibiting
adenosine metabolism and T-cell activation
Systemic or Non-organ Specific
B. Systemic Lupus Erythematosus
HYPERSENSITIVITY TYPE 3
● Chronic, inflammatory multi-organ disorder
that predominantly affects young women of
childbearing age.
The peak age of onset is usually between 20 and 40
years. Women are much more likely to be affected
than men, by a ratio of about → 9 to 1.1
● Immune complexes are formed → in SERUM
●
● Immune complexes are TRAPPED in →
basement membrane of glomeruli,
skin/endothelium, synovial joints, kidney
●
● Anti-phospholipid antibodies are present
SLE - genetically predisposed in oatients that have
HLADR2 & HLADR3
ETIOLOGY OF SLE
● Environmental factors thought to play a role
in SLE include → UV light, certain medications,
and possibly infectious agents.
 IMMUNOLOGY AND SEROLOGY
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SLE appears to → originate from complex interactions
between environmental factors, genetic susceptibility,
and abnormalities within the immune system.
→ Exposure to sunlight is well trigger of the
photosensitive skin rashes seen in many lupus and
also hormones as indicated by the significantly higher
incidence of lupus in women than in men.
● After joint involvement, the next most
common signs of SLE are → skin
manifestations
●
● An erythematous rash may appear on any
area of the body exposed to UV light.

GENETIC MAKE UP could also play an important role ● Less common but perhaps more dramatic is
as people with certain → HLA TYPES esp. → HLA DR3 the classic butterfly rash across the nose and
and HLA DR2 cheeks that appears in some SLE patients

HLA A1 and B8 → have INCREASE CHANCE of After joint involvement, the next most common signs
developing LUPUS are → skin manifestations. These can present in
various forms and are experienced by about 80% of
● Hormones are also important as indicated by
patients with lupus.
the significantly higher incidence of lupus in
females and an increased risk of developing This rash is responsible for the name lupus, derived
lupus in women that have used from the Latin term meaning “wolf-like.”
estrogen-containing contraceptives or
→ Evidence of renal involvement is present in about
hormone replacement therapy.
half of all patients with lupus; nephritis is a → major
●
cause of illness and death.
● Genetic makeup is believed to play an
important role in susceptibility to SLE There are several types of lesions, but the most
dangerous is → diffuse proliferative
Clinical Significance
glomerulonephritis, characterized by cell proliferation
● Autoantibodies associated with SLE include in the glomeruli that can lead to end-stage renal
antibodies to double stranded DNA (dsDNA), disease.
histones, and other nuclear components, as
● Evidence of renal involvement is present -
well as autoantibodies to lymphocytes,
nephritis is a major cause of illness and death
erythrocytes, platelets, phospholipids,
●
ribosomal components, and endothelium.
● Other systemic effects may include cardiac
●
involvement with pericarditis, tachycardia, or
● Anti-dsDNA and complement proteins have
ventricular enlargement; pleuritic with chest
been found in immune complexes that are
pain; and neuropsychiatric manifestations
deposited in organs such as the kidneys and
skin and are thought to play a major role in
the → pathogenesis of SLE.
●
● Accumulation of IgG to dsDNA forms
complexes of an intermediate size that
become deposited in the glomerular
basement membrane (KIDNEYS).

70% of patients that have lupus have antibodies to

→ DOUBLE STRANDED DNA

dsDNA → is regarded as HIGHLY SPECIFIC for SLE

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Positive Patterns observed under fluorescence

microscope

Immunofluoresce Antibody involved Autoimmune

nt pattern disease

Homogenous Anti-nucleoprotein RA Chronic active

Fluoresence hepatitis Sjogren’s
(staining of the Syndrome
entire nucleus) Drug-induced SLE
Myasthenia gravis

Membranous/ Anti-DNA/native SLE

Shaggy/ Peripheral DNA (double
Staining (staining stranded)
Methods of ANA Detection around the nuclues) Anti-nuclear antigen
→ Indirect Immunofluorescence (IIF) Fluorescent
Speckled Non-DNA nuclear SLE Scleroderma
antinuclear antibody (FANA) testing fluoresence constituents/ Mixed connective
(numerous minute anti-ENA tissue disease
Indirect Immunofluorescence (IIF) → Fluorescent fluorescent points (extractable nuclear Sjogren’s syndrome
antinuclear antibody (FANA) testing has been the throughout the antigen) a. Anti-Sm
nucleus) b. Anti-RNP
most widely used and accepted test because it is
highly sensitive, detects a wide range of antibodies, Nucleolar Some Anti-RNA SLE Scleroderma
and is inexpensive and easy to perform. fluoresence only Anti-fibrillarin
(fluoresence on the
Fluorescent antinuclear antibody (FANA) testing nucleus only)

Fine Speckled Anti-centromere CREST syndrome (a

pattern of antibody variant of SLE)
fluoresence

Interpretation:

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● Discrete and speckled pattern

● Antibodies to centromeric chromatin of
Homogenous (solid, diffuse)
metaphase and interphase cells
● Whole nucleus fluoresces evenly → gold ● Highly selective of CREST variant of
● It is associated with antibodies to dsDNA (also progressive systemic sclerosis
known as native or nDNA), histones, and ● Infrequently found in → SLE and other MCTD
deoxyribonuclear protein (DNP).
● The homogenous pattern is found in patients
with SLE, drug-induced lupus, and many other
autoimmune diseases

Peripheral (rim, outline, membranous, shaggy,

thready)

● Sharp, ring-gold fluorescence of outer edge on

nucleus with a gradually darkening inner
border blending with a dark nuclear center
● Antibodies to nDNA, dsDNA
● Seen in active stage of → SLE
● This pattern is primarily caused by antibodies
to dsDNA and is highly specific for SLE.

Speckled (mottled)
● Numerous round speckles of green-gold
nucleoli of various sizes against a dark
background; “pepper dots”
● Antibodies to ENA (Sm and RNP)
● Anti-RNP is indicative of other rheumatic
diseases
● The speckled pattern is associated with
antibodies to ENAs and can be found in
patients with SLE, Sjögren’s syndrome,
systemic sclerosis, and other systemic
autoimmune rheumatic diseases
Nucleolar
● Multiple, round, smooth, green-gold
fluorescing nucleoli of various sizes
● Antibodies to nRNA
● The nucleolar pattern is primarily caused by
antibodies to RNA and RNP and is seen
mainly in patients with scleroderma, but can
also be present in patients with other
connective tissue diseases
Anti-centromere antibody (ACA)
Systemic or Non-organ Specific
C. Scleroderma
● Primarily in skin/ectodermal tissues, but can
cause multi-organ pathology
● Skin fibroblast reproduce → faster and
produce more collagen
General signs of autoimmune diseases that may have
diagnostic value
1. Elevated → serum gamma globulins
2. Presence of diverse antibodies
3. Depressed levels of serum complement
4. Immune complexes in serum
5. Depressed levels of T cells
6. Lesions detected on biopsy resulting from
deposition of immune complexes
Other Facts about Autoimmune Diseases
1. Patient may have one autoantibody and in
fact, may suffer form multiple autoimmune
diseases
2. Although SLE is associated primarily with
anti-nuclear antibodies and rheumatoid

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arthritis primarily with Rf; both types of

antibodies amy be found in BOTH diseases
3. Autoantibodies are NOT unique to
autoimmune diseases
4. Autoimmune diseases are also grouped into
two:

a. Humoral or antibody-mediated

b. Cell-mediated or T-cell mediated

Laboratory Diagnosis of Autoimmune Diseases

1. ELISA
2. Indirect Immunofluorescence (IF)
3. RIA
4. Immunoblotting
5. Immunodiffusion, Binding Assays

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PRINCIPLES OF IMMUNOLOGIC AND ● Either you are detecting the patient antigen or
SEROLOGIC PROCEDURES antibody and → the reagent will consist either
the antigen or antibody.
Learning Intended Outcomes
● Explain the principles and apply the Antigen–Antibody Binding
procedures of the different serologic tests in
the detection of antigens and antibodies. The primary union of binding sites on an antibody
● Distinguish between precipitation and with specific epitopes on an antigen depends on two
agglutination reaction correctly. characteristics of antibody known as AFFINITY and
● Compare and describe the different AVIDITY.
agglutination reactions and give example of
For such reactions to occur or for an antigen antibody
each appropriately.
to occur both antigen and antibody must have
● Compare and contrast single and double
MULTIPLE BINDING SITE for them to from a LATTICE
immunodiffusion discuss the role of each in
FORMATION of CROSSLINK → and the binding
measurement of precipitation reactions. characteristics now of antibodies are termed as
● Compare flocculation and agglutination AFFINITY and AVIDITY.
● Explain the principles of complement fixation
and neutralization test accurately. AFFINITY and AVIDITY - plays MAJOR role in antigen
and antibody binding or reaction,
Antigen–Antibody Binding
AFFINITY
In serology the focus is to → determine the absence
● INITIAL FORCE OF ATTRACTION that exists
or presence of an antibody or an antigen to help in the
between a single Fab site on an antibody
diagnosis of certain infections and diseases
molecule and a single epitope or determinant
site on the corresponding antigen
The combination of an antigen with a specific
● The strength of attraction DEPENDS on the
antibody plays an important role in the laboratory in
specificity of antibody for a particular antigen
diagnosing many different diseases and there are a lot
An antigen has different antigenic determinants.
of SEROLOGICAL TEST or IMMUNOASSAYS.
Antigenic determinants or epitope - is the binding site
of the antibody molecule.
IMMUNOASSAYS - we're using ANTIBODIES in the
Cross reaction mechanism - wherein an antigen can
diagnosis of different diseases.
have several antigenic determinants and antibody can
react to this.
there are several immunoassays that have been
developed to detect (sa isang immunoassays we’re
eg: antibody of smallpox pwedeng mag react sa
detecting either the ANTIGEN or ANTIBODY of a
antigenic determinant ng cowpox and that is a cross
particular INFECTION, and kung ano yung denedetect
reaction mechanism but the affinity increases if the
ang reagent is yung OPPOSITE niya)
particular antibody is for the particular antigen (so
mas malakas ang affinity ni cowpox antibody to the
If there is an ANTIGEN ANTIBODY BINDING there
cowpox antigen kesa cowpox antibody to the smallpox
would be a particular reaction to help diagnose a
antigen)
particular infection.
The more cross-reacting antigen resembles the
If you are detecting patients antigen the reagent is
original antigen the STRONGER the bond will be
consist of the KNOWN ANTIBODY for it (para mag
between the antigen and the binding site of an
positive at ma detect yung certain infection)
antibody

So if the epitope or the antigenic determinants and

the binding site of an antibody exhibits a perfect lock
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and key that is → the case of of an original antigen and
the affinity will be at its maximum or maximum
capacity
One antibody may initially attract numerous different
antigen but the epitopes shape and the way it fits
together with binding site of an antibody determines
whether the bonding will be stable or not.
Antibodies are capable of reacting with antigens
resembling the original antigen that induced antibody
production, which is known as → cross-reactivity.
The more the cross-reacting antigen resembles the
original antigen, the stronger the bond will be
between the antigen and the binding site. However, if
the epitope and the binding site have a perfect
lock-and-key fit, as is the case with the original
antigen, the affinity will be maximal.
When the affinity is higher, the assay reaction is more
sensitive because more antigen–antibody complexes
will be formed and visualized more easily.
● When the affinity is higher, the assay reaction
is more sensitive because more
antigen–antibody complexes will be formed
and visualized more easily.
FIGURE 10–1 Affinity is determined by the
three-dimensional fit and molecular attractions between
one antigenic determinant and one antibody-binding site.
The antigenic determinant on the left has a better fit and
charge distribution than the epitope on the right and hence
will have a higher affinity.
AVIDITY
● SUM of all attractive forces between an
antigen and an antibody
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● This involves the strength with which a
multivalent antibody binds a multivalent
antigen,
Avidity → represents the overall strength of
antigen–antibody binding and is the sum of the
affinities of all the individual antibody–antigen
combining sites.
Avidity → refers to the strength with which a
multivalent antibody binds a multivalent antigen and
is a measure of the overall stability of an
antigen–antibody complex.
In other words, once binding has occurred, it is the
force that keeps the molecules together
● A measure of the overall stability of an
antigen– antibody complex.
● A high avidity can actually compensate for a
low affinity.
The more bonds that form between antigen and
antibody, the higher the avidity is. IgM → for instance,
has a higher avidity than IgG because
IgM - has the potential to bind 10 different antigens
Both affinity and avidity → contribute to the stability
of the antigen–antibody complexes,which is essential
to detecting the presence of an unknown, whether it
is antigen or antibody.
● Stability of the antigen–antibody complex is
essential to detecting the presence of an
unknown, whether it is antigen or antibody
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Avidity is the sum of the forces binding multivalent
antigens to multivalent antibodies. In a comparison between
IgG and IgM, IgM has the most potential binding sites for
antigen and thus the higher avidity. Note that the monomers in
IgM can swing up or down in order to bind more effectively.
Law of Mass Action
All antigen–antibody binding is REVERSIBLE and is
governed by the → LAW OF MASS ACTION
● This law states that free reactants are in
equilibrium with bound reactants
● The equilibrium constant K represents the
difference in the rates of the forward and
reverse reactions according to the following
equation:
▹ K = [AgAb]/[Ab][Ag]
(Equilibrium constant is equals to antigen-antibody
association / antibody and antigen dissociation
complex)
■ where [AgAb] = concentration of
antigen-antibody complex (mol/L)
■ [Ab] = concentration of free antibody (mol/L)
■ [Ag] = concentration of free antigen (mol/L)
As the strength of binding, or avidity, increases,
the tendency of the antigen–antibody complexes to
dissociate decreases, which increases the value of K.
When the value of K is higher, the amount of
antigen–antibody complex is larger and the assay
reaction is more visible or easily detectable.
HIGHER AVIDITY = HIGHER ASSOCIATION or BINDING
HIGHER AVIDITY = LOW DISSOCIATION
The ideal conditions in the clinical laboratory would
be to have an antibody with a high affinity, or initial
force of attraction, and a high avidity, or strength of
binding.
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The higher the values are for both of these and the
more antigen–antibody complexes that are formed,
the more sensitive the test.
Precipitation and Agglutination
Precipitation and Agglutination are the visible
expression of the aggregation of antigens and
antibodies through the formation of a framework in
which antigen particles or molecules alternate with
antibody molecules.
PRECIPITATION - this involves combination of a
soluble antigen with a soluble antibody (so pareho
dapat soluble antigen when it comes to precipitation
reactions)
PRECIPITATION - this involves a combination of a
soluble antigen and then the soluble antibody to
→ produce an insoluble complex which are visible
AGGLUTINATION - this is the process by which the
antigen is PARTICULATE such as particles → such as for
example cells which aggregate to form a large complex
when a specific antibody is present or if the antibody
the
is bound to the particulate antigen.
Precipitation and agglutination - these are the visible
expression of the aggregation of antigen and
antibodies through the formation of a framework
So for a visible reaction or expression of an antibody
complex occur or the antigen and antibodies should
produce a LATTICE FORMATION or a FRAMEWORK in
which the particles or the molecules alternate with
the antigen rather alternate with the antibody
molecules.
PRECIPITATION REACTIONS
Precipitation Curve
PRECIPITATION - this involves a combination of a
soluble antigen and then the soluble antibody to
→ produce an insoluble complex which are visible
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Precipitation - depends on the relative proportions of
antigen and antibody present in the reaction.
Optimum precipitation should be in the zone of
equivalence.
ZONE OF EQUIVALENCE
● In this zone, precipitation is the result of
random, reversible reactions whereby each
antibody binds to more than one antigen and
vice versa, forming a stable network or lattice
In the zone of equivalence, the number of multivalent
sites of antigen and antibody are approximately equal
(para magkaroon ng stable network or LATTICE
FORMATION)
The lattice hypothesis, as formulated by Marrack, is
based on the assumptions that each antibody
molecule must have at least two binding sites and the
antigen must be multivalent.
As they combine, this arrangement results in a
multimolecular lattice that increases in size until it
precipitates out of solution
Prozone phenomenon
The prozone phenomenon occurs, in which antigen
combines with only one or two antibody molecules
and no cross-linkages are formed.
In the prozone, usually only one site on an antibody
molecule is used and many free antibody molecules
remain in solution.
● Antigen combines with only one or two
antibody molecules and no cross-linkages are
formed.
● This is because usually only one site on an
antibody molecule is used, and many free
antibody molecules remain in solution
Postzone phenomenon
● At the other side of the zone, where there is
antigen excess, the postzone phenomenon
occurs in which small aggregates are
surrounded by excess antigen, and again no
lattice network is formed.
● In this case, every available antibody site is
bound to a single antigen, and no cross-links
are formed.
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● Thus, for precipitation reactions to be
detectable, they must be run in the zone of
equivalence
FIGURE 10–3 Precipitin curve. The precipitin curve shows
how the amount of precipitation varies with varying antigen
concentrations when the amount of antibody is kept
constant. Excess antibody is called the prozone and excess
antigen concentration is called the postzone.
The prozone and postzone phenomena must be
considered in the clinical setting because negative
reactions occur in both.
A false-negative reaction may take place in the
prozone because of high antibody concentration.
If it is suspected that the reaction is a false negative,
diluting out antibody and performing the test again
may produce a positive result. In the postzone,excess
antigen may obscure the presence of a small amount
of antibody.
Typically, such a test is repeated with an additional
patient specimen taken about a week later.
The extra time would allow for the further production
of antibody . If the repeated test is negative, it is
unlikely that the patient has that particular antibody.
3 test for precipitation
● Precipitation by Light Scattering
● Precipitation by Immunodiffusion/ diffusion
into the gel
● Precipitation by using electric current /
electrophoresis
A. Precipitation by Light Scattering
1. Turbidimetry
- directly through the solution
● a MEASURE of the turbidity or cloudiness of a
solution
because most antigens are multivalent and thus
capable of → forming aggregates in the presence of
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the corresponding antibody. When antigen and
antibody solutions are mixed, the antigen cross-links
with numerous antibody molecules and the lattice
networks become so large that they precipitate out of
solution.
Precipitates in fluids can be measured by means of
→ turbidimetry or nephelometry.
● A detection device is placed in direct line with
the incident light, collecting light after it has
passed through the solution.
● Scattering of light occurs in proportion to the
size, shape, and concentration of molecules
present in solution.
● Measurements are made using
spectrophotometer or an automated clinical
chemistry analyzer
2. Nephelometry
- gumagamit ng certain angle
- Nephelometers typically measure light scatter
at angles ranging from 10 degrees to about 90
degrees.
● measures the light that is scattered at a
particular angle from the incident beam as it
passes through a suspension
● If a solution has excess antibody, adding
increasing amounts of antigen results in an
increase in antigen–antibody complexes and
thus an increase in light scattering.
● Nephelometry can be used to detect either
antigen or antibody, but typically it is run with
antibody as the reagent and patient antigen as
the unknown
● Although the sensitivity of turbidity has
increased, nephelometry is more sensitive,
with a of detection of 1 to 10 mg/L for serum
proteins
● The amount of light scattered is an index of
the solution’s concentration
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FIGURE 10–4 Principles of nephelometry. The light
detection device is at an angle to the incident light, in
contrast to turbidity, which measures light rays passing
directly through the solution.
a
B. Precipitation by Passive Immunodiffusion
PRECIPITATION - always dealing with SOLUBLE
ANTIGEN and SOLUBLE ANTIBODY that precipitates
out of the serum forming an → INSOLUBLE COMPLEX
When the number of antigen and antibody is EQUAL
the ZONE OF EQUIVALENCE will occur resulting to a
VISIBLE REACTION
● Antigen and antibody are added to wells in
the gel (AGAROSE) an antigen– antibody
combination occurs by means of diffusion
➔ Agarose, a purified high-molecular-weight
complex polysaccharide derived from
seaweed, is used for this purpose.
➔ When antigen and antibody diffuse toward
one another in a gel matrix, visible lines of
precipitation will form.
➔ Agarose - helps stabilize the diffusion process
and allow visualization of the precipitin bands
(that’s why yun yung ginagamit na support
medium in immunodiffusion)
● No electrical current is used to speed up the
process
● The rate of diffusion is affected by the size of
the particles, the temperature, the gel
viscosity, and the amount of hydration.
The agar concentration that is used as a support
medium in this reaction ranges from → 0.3% - 1.5% of
agar and concentration allows for diffusion of most
reactants
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TECHNIQUES: and this can be compared to a

○ Radial Immunodiffusion standard curve obtained by using
- single immunodiffusion antigens of known concentration
○ Ouchterlony Double Diffusion
- double immunodiffusion Radial Immunodiffusion (RID)
same gumagamit ng agarose gel and then hahayaan
mag diffuse ang antigen at antibody towards one
another in a gel matrix and if there is a reaction or
combination visible lines of precipitation will form in
the gel → PASSIVE DIFFUSION

- When no electrical current is used to speed up

this process, it is known as passive
immunodiffusion. The rate of diffusion is
affected by the size of the particles, the
temperature, the gel viscosity, and the
amount of hydration.

1. Radial Immunodiffusion (RID)
● James Oudin
● A single-diffusion technique (meaning
isang antigen isang antibody) detect is
the ANTIGEN OF THE PATIENT
In this technique, antibody is uniformly distributed in
the support gel and antigen is applied to a well cut
into the gel. As the antigen diffuses out from the well,
antigen – antibody combination occurs in changing
proportions until the zone of equivalence is reached
and a stable lattice network is formed in the gel. The
area of the ring obtained is a measure of antigen
concentration that can be compared with a standard
curve obtained by using antigens of known
concentration, depicts some typical results.
● Antibody is uniformly distributed in
support gel, and antigen is applied to
a well cut into the gel
● The area of the ring obtained is a
measure of antigen concentration,
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Antibody is incorporated into the agar and then layer
on top the antigen as the antigen moves down
towards the gel if present siya into particular antibody
a precipitation reaction will occur at the zone of
equivalence forming lattice formation and because of
this magkakaroon precipitation bond that resembles a
RING that’s why this is called → Radial
Immunodiffusion and the diameter of the ring will be
compared to a standard concentration and the
proportions of the ring is DIRECTLY PROPORTIONAL to
the antigen concentration
(so, compare yung diameter nung RING to the
STANDARD of KNOWN antigen concentration for us to
known the ANTIGEN CONCENTRATION OF UNKNOWN)
RID have 2 types of measurements
● A. Mancini/ Endpoint Method
● Fahey and McKelvey/ Kinetic Method
same lang na ginagamit na RID pero ang difference is
the TIME that is required to measure the ring or the
precipitating ring
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● Both antigen and antibody diffuse

MEAUSRING RADIAL IMMUNODIFFUSION
A. Mancini/ Endpoint Method
- PASSIVE IMMUNODIFFUSION TECHNIQUE
- measure at the END
One technique for the measurement of radial
immunodiffusion was developed by Mancini and is
known as the endpoint method.
Equivalence occurs between 24 and 72 hours.
● IgM - 50-72 hours
● IgG - 24 hours for the zone of equivalence to
occur
● Antigen is allowed to diffuse to completion
● Once equivalence is reached, no further
change in ring diameter takes place
● In this technique, antigen is allowed to diffuse
to completion, and when equivalence is
reached, there is no further change in the ring
diameter.
● The square of the diameter is then directly
proportional to the concentration of the
antigen.
● The major drawback to this method is the
time it takes to obtain results
independently through a semisolid medium in
two dimensions
In this technique, both antigen and antibody diffuse
independently through a semisolid medium in two
dimensions, horizontally and vertically.
● Precipitin lines form where the moving front
of antigen meets that of antibody.
● The density of the lines reflects the amount of
immune complex formed
Wells are cut in a gel and reactants are added to the
wells.
- Antibody that is multispecific is placed in the
central well and different antigens are placed
in the surrounding wells to determine if the
antigens share identical epitopes.
- Diffusion takes place radially from the wells.
After an incubation period of between 12 and
48 hours in a moist chamber, precipitin lines
form where the moving front of antigen meets
that of antibody and the point of equivalence
is reached.

B. Fahey and McKelvey/ Kinetic Method

- measure while the test is being conducted
● Uses measurements taken before the point of Ouchterlony Double Diffusion
equivalence is reached
the kinetic or Fahey method, uses ring diameter
readings taken at about 19 hours before equivalence
is reached
● The diameter is proportional to the log of the
concentration
RID - Radial immunodiffusion has been used to
measure IgG and IgA subclasses as well as
complement components.

B. Precipitation by Passive Immunodiffusion

2. Ouchterlony Double Diffusion

- Ouchterlony method
One of the older, classic immunochemical techniques
is Ouchterlony double diffusion

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● Diffusion can be combined with electrical

current to speed up reaction and sharpen
results
● Electrophoresis separates molecules according
to differences in their electric charge when
they are placed in an electric field.
● A direct current is forced through the gel,
causing antigen, antibody, or both to migrate.
As diffusion takes place, distinct precipitin
bands are formed.

TECHNIQUES
○ Rocket Immunoelectrophoresis
○ Immunoelectrophoresis
○ Immunofixation Electrophoresis
1. Rocket immunoelectrophoresis
is a one dimension electroimmunodiffusion, this is a
adaptation of the regional immunodiffusion or RID
FIGURE 10–6
which is developed by Laurel, same principle but this
Ouchterlony diffusion patterns. An antibody mixture is
placed in the central well. Unknown antigens are placed
is combined with electrical current to speed up
in the outside wells. The antibodies and antigens all reaction and sharpen results
diffuse radially out of the wells. - it just RID + electrophoresis
● Antibody is distributed in the gel, and antigen
(A) Serological identity. If the antigens are identical, they is placed in wells cut in the gel, just as in RID.
will react with the same antibody and the precipitate ● Electrophoresis is used to facilitate migration
line forms a continuous arc. of the antigen into the agar
(B) Nonidentity. If the antigens share no identical
● When the antigen diffuses out of the well,
determinants, they will react with different antibodies
and two crossed lines are formed.
precipitation begins.
(C) If antigen 3 has a determinant in common with ● The end result is a precipitin line that is
antigen 1, one of the antibodies reacts with both conical in shape, resembling a rocket, hence
antigens. the name rocket immunoelectrophoresis.
Another antibody that reacts with different determinants
on antigen 1 (absent on antigen 3) passes through one
precipitation line and forms the spur on the other line.
The spur formed always points to the simpler antigen
with fewer antigenic determinants.
The position of the precipitin bands between wells
allows for the antigens to be compared with one
another.
Several patterns are possible:
(1) Fusion of the lines at their junction to
form an arc represents serological identity or the
presence of a common epitope,
(2) a pattern of crossed lines demonstrates two
separate reactions and indicates that the compared
antigens share no common epitopes, and
(3) fusion of two lines with a spur indicates partial
identity.

C. Precipitation by Electrophoretic Techniques

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2. Immunoelectrophoresis
introduced by GRAY BAR and WILLIAMS GRAY
● A double-diffusion technique that
incorporates electrophoresis current to
enhance results
● A trough is cut in the gel parallel to the line of
separation
● Antiserum is placed in the trough
● Incubated time: 18 to 24 hours.
double-diffusion occurs at right angles and
electrophoresis will be used to separate the reactions
if there is presence of antigens towards the antibodies FIGURE 10–7 Immunofixation electrophoresis. A
in the trough precipitin lines or bond will formed, complex antigen mixture such as serum proteins is
where the specific antigen antibody combination took separated by electrophoresis. An antiserum
place. template is aligned over the gel. Then protein
- However, interpretation is difficult for fixative and monospecific antisera, IgG, IgA, IgM, κ,
immunoelectrophoresis therefore it has and λ are applied to the gel. After incubating for 30
largely been replaced with immunofixation minutes, the gel is stained and examined for the
electrophoresis give faster and more reliable presence of paraproteins. Precipitates form where
results and is easier to interpret specific antigen–antibody combination has taken
3. Countercurrent Immunoelectrophoresis place. In this case, the patient has an IgG
same as immunoelectrophoresis but instead of a monoclonal antibody with λ chains. (Courtesy of
trough Helena Laboratories, Beaumont, TX.)
● Ag and ab placed on a well directly opposite
each other
use to facilitate migration towards the center SUMMARY
(magkikita sa gitna)
Precipitin line is formed it will indicate that there has
been a reaction, so precipitin line closer to the antigen
well it means that there is a higher concentration
antibody
4. Immunofixation Electrophoresis
immunoelectrophoresis this was describe by ALPER
and JOHNSON
● Similar to immunoelectrophoresis except that
after electrophoresis takes place, antiserum is
applied directly to the gel’s surface rather
than placed in a trough
● Immunodiffusion takes place in a shorter time
(less than 1 hour) and results in a higher
resolution AGGLUTINATION REACTIONS (PART 2)
● Agarose or cellulose acetate is used.
● An antibody of known specificity is used to Agglutination
determine whether patient antigen is present. Agglutination - it needs that the antigen is a
PARTICULATE MATTER kasi ang SOLUBLE ANTIGEN
kahit na may combination sila with an antibody hindi
sila mag formed ng visible agglutination
● Agglutination (clumping) is the term used to
describe the aggregation of particulate test
antigens.

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● Process by which particulate antigens such as

cells aggregate to form large complexes when
a specific antibody is present.
● Agglutination occurs only if the antigen is in
the form of particles such as bacteria, red
blood cells, latex particles, white blood cells
or any substance that appears cloudy when
suspended in saline. Lifted from: Stevens
In 1896, Gruber and Durham published the first report
about the ability of antibody to clump cells, based on
observations of agglutination of bacterial cells by
serum.
Sensitization
● involves antigen–antibody combination
through single antigenic determinants on the
particle surface
Lattice formation
● Formation of cross-links that form the visible
aggregates.
● This represents the stabilization of
antigen–antibody complexes with the binding
together of multiple antigenic determinants

This finding gave rise to the use of serology as a tool in Phases of Agglutination
the diagnosis of disease and also led to the discovery
of the ABO blood groups.

Widal and Sicard developed one of the earliest

diagnostic tests in 1896 for the detection of antibodies
occurring in typhoid fever, brucellosis, and tularemia.

➔ Direct agglutination
➔ Passive Agglutination
➔ Reverse Passive Agglutination
➔ Agglutination Inhibition
➔ CO agglutination
➔ Antiglobulin mediated immune agglutination FIGURE 10–8 Phases of agglutination. Sensitization:
Antigen and antibody unite through antigenic
Mechanism of particle agglutination determinant sites. Lattice formation:
Agglutination is the clumping aggregation of particles Rearrangement of antigen and antibody bonds to
that's why it involves particulate antigens form a stable lattice.
It involves clumping or aggregation of particles that Sensitization is affected by the nature of the antigens
have antigens on its surface and combining with on the agglutinating particles. If epitopes are sparse or
antibody molecules that form bridges between the if they are obscured by other surface molecules, they
antigenic determinants of the antigens are less likely to interact with antibody.
Agglutination, like precipitation, is a two-step process
Additionally, red blood cells (RBCs) and
that results in the formation of a stable lattice
bacterial cells have a slight negative surface charge;
network.
because like charges tend to repel one another, it is
➔ The first reaction, called sensitization,
sometimes difficult to bring such cells together into a
involves antigen–antibody combination
lattice formation.
through single antigenic determinants on the
particle and is rapid and reversible.
- The class of immunoglobulin is also important; IgM
➔ The second step, or lattice formation, is the
with a potential valence of 10 is over 700 times more
formation of cross-links that form the visible
efficient in agglutination than is IgG with a valence of
aggregates. This represents the stabilization of
Antibodies of the IgG class often cannot bridge the
antigen–antibody complexes with the binding
distance between particles because their small size
together of multiple antigenic determinants
and restricted flexibility at the hinge region may
prohibit multivalent binding.
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IgM antibodies, on the other hand, are strong Hemagglutination:

agglutinins because of their larger size. Achieving
visible reactions with IgG often requires the use of
enhancement techniques that vary physicochemical
conditions such as the ionic strength of the solution,
the pH, and the temperature.
Antibodies belonging to the IgG class agglutinate
best at 30°C to 37°C, whereas
IgM antibodies react best at temperatures between
4°C and 27°C.
Because naturally occurring antibodies against the
ABO blood groups belong to the IgM class, these
reactions are best run at room temperature.
Antibodies to other human blood groups usually
belong to the IgG class; reactions involving these must
be run at 37°C
● Antigen is found on surface of RBCs
If an agglutination reaction involves RBCs, then it is
called hemagglutination.
- The best example of this occurs in ABO blood
group typing of human RBCs, one of the
world’s most frequently used immunoassays.
- Patient RBCs mixed with antisera of the IgM
type can be used to determine the presence
or absence of the A and B antigens; this
reaction is usually performed at room
temperature without the need for any
enhancement techniques.
Hemagglutination Reactions

In addition to temperature considerations, detection

of IgG antibodies often requires the use of a second
antibody, antihuman immunoglobulin, to visualize a
reaction

TYPES OF AGGLUTINATION REACTONS

1. DIRECT AGGLUTINATION
● Occurs when antigens are found naturally on
the surface of a particle.
● One of the best examples of direct
agglutination testing involves known bacterial
antigens used to test for the presence of FIGURE 10–11 Grading of agglutination reactions:
unknown antibodies in the patient. A. tube method. If tubes are centrifuged and shaken
● Detection of antibodies is primarily used in to resuspend the button, reactions can be graded
diagnosis of diseases for which the bacterial from negative to 4+, depending on the size of clumps
agents are extremely difficult to cultivate. observed. B. Rapid slide method.
One such example is the Widal test, a rapid screening
test used to help determine the possibility of typhoid
fever.
A significant finding is a fourfold increase in antibody
titer over time when paired dilutions of serum
samples are tested with any of these antigens.

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FIGURE 10–10 Red blood cell agglutination. The tube
on the left is a positive test for RBC agglutination,
whereas the tube on the right is a negative test
showing that the RBCs have remained in a smooth
suspension.
2. PASSIVE AGGLUTINATION
gumagamit here ng carrier
A variety of particles, including erythrocytes, latex,
and gelatin, are used for passive agglutination.
● Passive, or indirect, agglutination employs
particles that are coated with antigens not
normally found on their surfaces.
● Particle sizes vary from 7 μm for RBCs down to
0.8 μm for fine latex particles
● Used to detect rheumatoid factor; antinuclear
antibody occurring in the disease lupus
erythematosus; antibodies to group A
streptococcus antigens; antibodies to
Trichinella spiralis; antibodies to Treponema
pallidum; and antibodies to viruses such as
cytomegalovirus, rubella, varicella-zoster,
and HIV-1/ HIV-2
Different carrier particles such as:
➔ commonly use is LATEX
➔ RBCs
➔ Gelatine
➔ Silicates
➔ Bentonite
➔ Charcoal
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PASSIVE AGGLUTINATION
In 1955, Singer and Plotz found by happenstance that
IgG was naturally adsorbed to the surface of
polystyrene latex particles.
- Latex particles are inexpensive, are relatively
stable, and are not subject to cross-reactivity
with other antibodies. A large number of
antibody molecules can be bound to the
surface of latex particles, so the number of
antigen-binding sites is large
Additionally, the large particle size facilitates reading
of the test. Latex agglutination tests have been used
to detect rheumatoid factor, antibodies to Group A
Streptococcus antigens, and antibodies to viruses
such as rotavirus, cytomegalovirus, rubella, and
varicella-zoster
3. REVERSE PASSIVE AGGLUTINATION
Same as passive agglutination meaning it also use a
carrier particles → the antigens used in this reaction
cannot form visible reaction that's why it needs or
have a carrier molecule but the difference of passive
to reverse passive agglutination -
Reverse passive - uses antibody rather than antigen so
the antibody is the one attached to the carrier to the
latex so the carrier molecule that's why if
agglutination occur patient antigen is present
(Kung anong reagent mo yung kabaliktaran non yun
yung detect)
So dahil reagent is carrier particles mixed with the
reagent antibody so carrier + antibody if there is
agglutination → it means there is presence of patient
antigen
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● Antibody rather than antigen is attached to a
carrier particle.
● The antibody must still be reactive and is
joined in such a manner that the active sites
are facing outward (so that presence and
available parin yung binding site)
● This type of testing is often used to detect
microbial antigens
Numerous kits are available for the rapid identification
of antigens from such infectious agents as Group B
Streptococcus, Staphylococcus aureus, streptococcal
groups A and B, rotavirus, and Cryptococcus
neoformans.
Passive and Reverse Passive Agglutination
FIGURE 10–12 Passive and reverse passive
agglutination. (A) Passive
agglutination. Antigen is attached to the carrier
particle; agglutination occurs if patient antibody is
present. (B) Reverse passive agglutination. Antibody
is attached to the carrier particle; agglutination
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occurs if patient antigen is present.
4. COAGGLUTINATION
This is just the same with reverse passive
agglutination but the difference the particle or the
carrier is bacteria, antibody ang naka attached
Because staphylococcus aureus has a protein on its
surface that we call protein A which naturally absorbs
like hinihigop niya naturally fc portion na antibody
molecules that's why staphylococcus aureus serve at
the carriers and observe the fc portion of antibodies
● Uses bacteria as the inert particles to which
antibodies are attached
● Staphylococcus aureus is most frequently
used
● These particles exhibit particles and are more
refractory to changes in ionic strength.
● Coagglutination reagents have been used in
identification of streptococci, Neisseria
meningitidis, Neisseria gonorrhoeae, Vibrio
cholera 0139, and Haemophilus influenzae
Protein A absorbs the fc portion of IgG except for
IgG3.
IgG3 - fails to attached to S.aureus Protein A
5. AGGLUTINATION INHIBITION
● Based on competition between particulate
and soluble antigens for limited
antibodycombining sites
● Lack of agglutination is an indicator of a
positive reaction
● Classic example is the human chorionic
gonadotropin (HCG)
Typically, this type of reaction involves haptens
that are complexed to proteins; the hapten–protein
conjugate is then attached to a carrier particle.
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TYPES OF AGGLUTINATION REACTONS

HEMEAGGLUTINATION INHIBITION
used RBCs
● Reactions use the same principle, except red
blood cells are the indicator particles
● Presence of patient antibody inhibits the
agglutination reaction
Agglutination inhibition reactions are based on
competition between particulate and soluble antigens
for limited antibody combining sites.

Hemagglutination inhibition reactions use the same

principle, except RBCs are the indicator particles.

This type of testing has been used to detect antibodies

to certain viruses, such as rubella, influenza, and
respiratory syncytial virus (RSV)

- The patient sample is first reacted with a

limited amount of reagent antibody that is
specific for the hapten being tested.
- Indicator particles that contain the same
hapten one wishes to measure in the patient
are then added.
- If the patient sample has no free hapten, the
reagent antibody is able to combine with the
carrier particles and produce a visible
agglutination. In this case, however,
agglutination is a negative reaction, indicating
that the patient did not have sufficient hapten 6. ANTIGLOBULIN-MEDIATED AGGLUTINATION
to inhibit the secondary reaction ● Detects non-agglutinating antibody which is
- Either antigen or antibody can be attached to IgG by means of coupling with a second
the particles. antibody which is known → antibody to
- The sensitivity of the reaction is governed by human globulin or anti-human globulin
the avidity of the antibody itself.
- It can be a sensitive assay capable of detecting ● The key component of the test is antibody to
small quantities of antigen. Tests used to human globulin that is made in animals or by
detect illicit drugs such as cocaine or heroin means of hybridoma techniques.
are examples of agglutination inhibition tests. IgG cannot bridge the gap of two antigen that's why
cannot form a stable lattice formation that's why kahit
na IgG nakabond sa kanyang antigen it cannot produce
a visible agglutination → IgG needs a second
antibody for an agglutination to occur and that
antibody is known as anti-human globulin

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Two different types of Coombs test:

B. INDIRECT ANTIGLOBULIN TEST
● Direct antiglobulin test - cross-matching
● Indirect antiglobulin test ● Detectin vitro sensitization
Coombs test - checks your blood for antibodies that ● Used to determine the presence of a
particular antibody in a patient, or it can be
attack red blood cells.
used to type patient red blood cells for
A. DIRECT ANTIGLOBULINTEST specific blood group antigens
● Used to demonstrate in vivo (in the body) Compatibility test - between the donor the recipient
attachment of antibody or complement to an
individual’s red blood cells.
● This test serves as an indicator of
autoimmune hemolytic anemia, hemolytic
disease of the newborn, sensitization of red
blood cells caused by the presence of drugs,
or a transfusion reaction

very important ang WASHING IN THIS TEST

Improper washing will produce false-negative

reaction because unbound antibody mga antibody na
di naka attach sa red cells can neutralized AHG

After that mag aadd na ng AHG kung present ang

patient antibody for this red cells magkakaroon ng
agglutination reaction ibig sabihin ng patient serum
may antibody that's why hindi pwede transfuse kasi
sisirain lang din niya yung red cells.

TYPES OF AHG REAGENT

a. Polyspecific AHG contains

● Anti IgG and anti C3d

b. Monospecific AHG contains

● Contain anti-IgG or anti-C3d

O Check Cells
- This use to validate a negative reaction kung
talagang negative ba o nag negative lang kasi
na neutralize yung AHG

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● Group O RBCs sensitized with IgG

● After addition of O check cells, agglutination
indicates:
■ AHG reagent was added
■ AHG reagent was not neutralized
● Lack of agglutination, invalidates results

Causes of False-Positive Reactions in Agglutination

Testing

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IMMUNOLOGY AND SEROLOGY OF BACTERIAL &

VIRAL INFECTIONS
IMMUNOLOGY AND SEROLOGY OF INFECTIOUS
DISEASE
(Bacterial, Viral, Fungal and Parasitic)
Serological and Molecular Detection of Bacterial
Infections
INTRODUCTION
Human Microbiome
● Collection of microorganisms that exists on
the body, such as bacteria, viruses, and
single-celled prokaryotic organisms (yeast and
fungi)
keeps us healthy in many ways and there commonly
known as NORMAL FLORA
They reside on our body. They get nutrients, vitamins
for our body but at the same time some of these
human microbiomes also protect us against disease
causing bacteria.
In digestion of our food they produce also certain
vitamins and they stimulate INNATE and ADAPTIVE
IMMUNE SYSTEMS
Infectious disease
● When a microbe causes damage to host cells
or altered physiology that results in clinical
signs and 3 symptoms of disease, the phrase “
There are also microbes that causes damages to host
cells or altered physiology that can result in clinical
signs and symptoms of disease and that is known as
→ INFECTIOUS ORGANISM
The establishment of an organism that leads to host
injury is referred to as an → “infection.” / infectious
disease
Human–Microbe Relationships
When an individual is born, a dynamic relationship
begins between the human host and the bacteria in
the environment.
Various bacteria establish themselves on the surfaces
of an individual, including the gastrointestinal tract,
creating a symbiotic relationship.
Symbiotic relationship - this is a living together of 2
unlike organism, in that case the bacteria or the
human microbiome and human body also known as
Indigenous microbiota (known as normal flora)
Indigenous microbiota
● Symbiotic bacteria that reside on and colonize
these host surfaces
The bacterial populations that exist on the body are
not homogenous; they vary in composition and
numbers, depending on the area of the body or the
part of the body that is invaded by those
microorganisms
For a microorganism to survive, the organism needs
to colonize the host and acquire nutrients.
Importantly, it must not stimulate the host’s immune
response (in the case of the indigenous microbiota) or
it must avoid or circumvent the immune responses.
Once established, it needs to be able to replicate and
disseminate to a preferred site in the body for survival
and eventually be transmitted to a new susceptible
host.
Three types of symbiotic relationships can exist
between humans and bacteria;
Commensalistic - No benefits, No harm
- In commensalistic relationships, there is no
apparent benefit or harm to either organism.
- An example of a commensalistic organism is
Staphylococcus epidermidis, which colonizes
and inhabits the human skin
- Staphylococcus epidermidis do not harm the
host pero wala din naman binibigay na benefit
to the host, so neutral lang ang organism
Neutral; No harm or benefit to either organism (EX.
Staphylococcus epidermidis and skin

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Mutualistic - normal flora which helps us
- In a mutualistic relationship, both humans
and the bacteria benefit.
- One example is the Lactobacillus species that
colonizes the epithelial surfaces of the vaginal
canal.
- The human host provides conditions
(temperature, atmosphere, nutrients) that
allow the bacteria to grow and multiply; in
exchange, the bacteria produce lactic acid to
help innate immune response, which prevents
colonization of bacteria and yeast that may
cause disease
both organisms benefit with each other (Ex.
Lactobacillus and epithelial surfaces of vaginal canal at
intestinal surfaces)
● Parasitic
- the encounter with specific organism
or viruses or bacteria are occasionally
results in harm to the host
- Parasite/bacteria or other organism
could get nutrients to the host
provide condition to allow grow and
multiplication
- while the parasite or the organism
causes harm to the host
- No benefits, nahaharm pa niya yung
host
In this case, a parasitic relationship exists between the
other organisms and the host. Establishment of an
organism that leads to host injury is referred to as an
infection.
Viruses, bacteria, or any parasite gets benefits and
causes harm 4 to the host
Infecting organism and host Interactions
Infectivity
● An organism’s ability to establish an infection
Infectivity - describes the proportion of individuals
exposed to a pathogen through horizontal
transmission (i.e., person-to person spread) who will
become infected.
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Another term that is used with similar meaning is
contagious.
For example, the measles virus is extremely
contagious and has a high degree of infectivity.
Pathogenicity
● Inherent capacity of an organism to cause
disease
Pathogenicity refers to the inherent capacity of an
organism to cause disease. This is a qualitative trait of
the organism determined by its genetic makeup.
Always symptomatic (APPARENT ILLNESS)
Although an organism may be pathogenic in nature, it
may not always cause disease.
The outcome of the host–pathogen interaction is
determined by several factors, including the host’s
immunologic status. Some microorganisms may only
cause disease or infection in individuals who have
compromised immune systems because of factors
such as chemotherapy, radiation therapy, or various
chronic diseases. These organisms are referred to as
“opportunistic pathogens.”
Virulence
● A quantitative trait that refers to the extent of
5 damage, or pathology, caused by the
organism
Severity of the reaction of the pathology produced
and measured in terms of FATALITY
For example,
Yersinia pestis, the causative agent of bubonic and
pneumonic plague, is considered to be extremely
virulent and is likely to cause severe illness and death
upon infection unless antibiotics are administered.
Immunogenicity
- ability to produce a specific immunity or the
infections has the ability to produce a specific
immunity
For example,
Measles virus is able to produce a lifelong immunity
to the host so once you get infected you will produce
antibodies or specific immunity against organism and
if that organism is able to produce specific immunity,
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meaning it is IMMUNOGENIC or it has a high
immunogenicity
Bacterial Virulence Factors
Bacterial properties or features that determine
whether an organism is pathogenic and able to cause
disease are referred to as “virulence factors.”
Factors that increase a bacterium’s virulence may be
classified as either structural components (e.g.,
endotoxin is a component of the cell walls of certain
bacteria) or as extracellular substances produced by
the bacteria, such as exotoxins.
Genetic determinants located on the bacterial
chromosome are generally responsible for the
production of structural or surface molecules, which
help the organism to attach to and colonize the host
MECHANISM OF EVASION
Evasion - ability of a certain organism to colonize
inside a host
● Structural components
● Genetic determinants
● Extracellular Virulence Factors
● Endotoxin and Exotoxins
● Extracellular Virulence Factors
Extracellular substances produced by bacteria also
contribute to an organism’s virulence by breaking
down primary or secondary defenses of the body,
damaging the host tissue and cells, or facilitating the
growth and spread of the organism.
Substances that perform the latter function are called
invasins. Several of the invasins include hyaluronidase,
collagenase, phospholipases, lecithinases, coagulase,
and various kinases.
● Endotoxin and Exotoxins
- produce by certain bacteria
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Bacteria may also produce two types of
toxins—endotoxin and exotoxins.
Endotoxin (lipid A) - is found in the LPS layer of the
cell walls of all gram-negative bacteria.
This results in inflammation, increased heart rate,
increased body temperature (fever), and a decrease
in blood pressure.
Unlike endotoxin, which has multiple effects on the
body, exotoxins have a very specific and targeted
activity.
Endotoxin has multiple effects on body and have a
very specific and targeted activity so they are protein
molecules that are released from a living bacteria and
are considered to be the most potent molecules
known to harm the to living organisms because they
can act as “superantigens” are NOT processed by
antigen presenting cells (APCs).
a “superantigen” induces activation of up to 20% of T
cells, resulting in a massive release of cytokines.
The systemic events brought on by the release of large
amounts of cytokines leads to what is referred to as
“toxic shock syndrome.”
Examples include the TSST-1 toxin produced by
Staphylococcus aureus and the superantigens
produced by S pyogenes, the cause of streptococcal
toxic shock syndrome (STSS).
Immune Defense Mechanisms
- How the body responds to certain microorganisms
Both innate and adaptive responses may occur after
an encounter with foreign antigens
Against potential pathogens the first line of defense is
natural immunity or structural barriers which
includes the Intact skin and mucosal surfaces
● Intact skin and mucosal surfaces
Antimicrobial defense peptides
○ Lysozyme, alpha defensin
Epithelial surface may have enzymes and nonspecific
antimicrobial defense peptides (ADPs) and proteins
that have antimicrobial activity.
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One example of an enzyme with specific antimicrobial extracellular products produced by bacteria such as
activity is lysozyme, which is found in many exotoxin.
secretions, including tears and saliva.
Cell-mediated immunity (CMI) - the other branch of
Lysozyme destroys the peptidoglycan found in the cell the adaptive immune response, is helpful in attacking
wall of bacteria, especially gram-positive bacteria. intracellular bacteria, such as Mycobacterium
tuberculosis, Legionella pneumophila, Listeria
One group of soluble peptides is the defensin
monocytogenes, and Rickettsia species.
peptides.
● Acute phase reactants
Defensins - are produced constitutively by the cells in
● Production of antibodies directed against
the body.
bacterial antigens
The three main classes of defensins are alpha, beta, ● Cell-mediated immunity (CMI)
and theta.
Mechanisms of Evasion
Alpha defensins - are produced by neutrophils, Ways of certain organism to grow, colonize and
certain macrophage populations, and Paneth cells of produce in a host.
the small intestine. This class of defensins is believed
Evade antibodies - – antigenic variation → coat
to disrupt the microbial membrane.
themselves with host’s protein or fibronectin to hide
Beta defensins - are produced by neutrophils as well their antigenic determinants
as epithelial cells lining the various organs, including
Block phagocytosis
the bronchial tree and genitourinary system. They are
believed to increase resistance of epithelial cells to Inactivate the complement cascade - c3b acts as
colonization. opsonin is inactivated
Theta defensins - are NOT found in humans. Immunologic Response to Several Important Bacteria
SYPHILIS
Antimicrobial proteins
● Genus Treponema contains four principal
Many antimicrobial proteins contribute to the innate species of pathogenic organisms:
immune response.
1. Treponema pallidum subspecies pallidum
For example,
● human syphilis
Complement proteins - can promote chemotaxis.
2. Treponema pallidum subspecies pertenue
Interleukins - are involved in the regulation of
● yaws
immune responses and inflammatory reactions.
3. Treponema pallidum subspecies endemicum
Prostaglandins are involved in the dilation and
constriction of blood vessels and modulation of ● Non venereal endemic syphilis
inflammation.
4. Treponema carateum
Leukotrienes are involved in inflammation and fever.
● Agent of pinta
Acute phase reactants also play important roles.
5. Treponema cuniculi
For example,
● Rabbit syphilis
C-reactive protein (CRP) - activates the complement
system and promotes phagocytosis by macrophages.
Adaptive immune responses - include the production
of antibodies directed against bacterial antigens or

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SYPHILIS
Mode of transmission:
● Sexual transmission
● Parenteral exposure through contaminated
needles and blood
● Congenital infections during pregnancy
● Blood tranfusion
The first diagnostic blood test for syphilis was the
Wassermann test (1906)
In the treatment of syphilis, heavy metals, such as
arsenic, were replaced by penicillin in the 1940s.
STAGES OF SYPHILIS
A. PRIMARY SYPHILIS (Early syphilis)
● Patient develops a characteristic, primary
inflammatory lesion called a chancre at the
point of initial inoculation and multiplication
of the spirochetes.
● Serological tests usually give non reactive
result (negative) at the time
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B. SECONDARY SYPHILIS
It is usually observed 1 to 2 months after the
disappearance of chancre
● usually observed 1 to 2 months after the
disappearance of chancre
● Systemic dissemination of the organism
occurs.
● Patients may develop Condylomata lata (flat
wart-like lesions)
Diagnosis:
● Darkfield microscopy of the fluid of the rash
● Serological tests
C. LATENT SYPHILIS
● Characterized by lack of clinical symptoms
● Patients are non-infectious at this time, except
pregnant women who can pass the infection
to the fetus
● Diagnosis can be made only by serologic
methods
D. TERTIARY OR LATE SYPHILIS
● Characterized by the appearance of lesions
called Gummas, CVD and Neurosyphilis
● Congenital syphilis occurs
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● Diagnosis is done by reactive serological test Syphilis Antigen

and a reactive spinal fluid test (Neurosyphilis)
→ VDRL using CSF (specimen of choice)
CONGENITAL SYPHILIS AND NEUROSYPHILIS
A. CONGENITAL SYPHILIS
● T.pallidum crosses the placenta from the 18th
of gestation
● Causes abortion or stillbirths; disfigurement,
blindness, deafness and other complications
Hutchinsonian Triad: triad of notched teeth, keratitis
and deafness
B. NEUROSYPHILIS
● Syphilis of the central nervous system
LABORATORY DIAGNOSIS
Diagnostic markers for Syphilis: ANTIBODIES
a. Reagin antibodies or non-treponemal antibodies
● Antibody-like substances which in theory are
the results of interaction of the Treponema
with host tissues.
● Non-specific antibodies of Treponema
because they are directed against the protein
antigen group common to pathogenic
spirochetes
● IgG and IgA types
b. Anti-treponemal antibodies (ATA)
● Specific antibodies developed in response to
T.pallidum infection.
● IgM in primary syphilis
● IgG type during secondary syphilis
LABORATORY DIAGNOSIS
Syphilis: ANTIGEN
a. Wasserman antigen
● Made up of cardiolipin which is isolated from
cardiac muscle
● A hapten
b. Treponemal antigen
● Reiter treponeme
○ non-virulent, cultivable variant of
T.pallidum
○ Commonly used in CF tests and TPI
● Nichol’s treponeme
○ Virulent strain of T.pallidum
○ used in fluorescent treponemal
antibody absorption (FTA-ABS)
LABORATORY DIAGNOSIS
Direct Detection
DIRECT DETECTION:
Direct observation of Organism
I. Darkfield Microscopy
● Darkfield condenser is used to keep all
incidental light out of the field except for that
captured by the organisms themselves
● Motility is a key to identification
● A negative test does not exclude a diagnosis of
syphilis.
II. Fluorescent Antibody Testing
● A sensitive and highly specific alternative to
darkfield microscopy.
● Live specimens are not required
LABORATORY DIAGNOSIS
Serological tests
A. Non Treponemal tests: VDRL & RPR
● Detect REAGIN Antibodies
● Not specific for syphilis
● Inexpensive to carry out
● Reagent: CLC
● Disadvantage: reagin disappears soon after
syphilis is cured
False-positive reactions:
Transient: Hepatitis, IM, Varicella, herpes, measles,
malaria, TB and pregnancy

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Sustained: SLE, Leprosy, IV drug use, Autoimmne
arthritis
Serological Tests
A. Non-Treponemal Test: VDRL
Venereal Disease Research Laboratories
Principle: floculation
Reagents:
Cardiolipin (0.03%)
○ Main reacting component
○ Alcoholic extract of normal beef heart
extract
Lecithin (0.21%)
○ Helps neutralize the
anti-complementary properties of
cardiolipin
Cholesterol (0.9%)
○ Provides adsorption centers
○ Increases the reacting surface of
cardiolipin
○ Increases the complement-fixing
capacity of cardiolipin with reagin
● 1% benzoic acid to stabilize the antigen at
6-10C
Procedure of VDRL
a. Qualitative Serum VDRL
● 50uL or 0.05mL serum
● 56C for 30 mins to inactivate complement
● One drop (1/60 mL) of VDRL antigen is added
to the ring
● Rotated for 4 minutes at 180rpm
● Determine presence of flocculation
○ Non-Reactive (NR)
■ No flocculation or no
clumping
○ Weakly Reactive (WR)
■ Slight flocculation (small
clumps)
○ Reactive
■ Definite flocculation (medium
and large clumps)
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b. Quantitative Serum VDRL
● Needle used shall deliver 75 drops of antigen
suspension per 1 mL
● 100 drops of saline per 1 mL
● The highest serum dilution that gives reactive
result is reported as the endpoint titer
● Often used as a therapeutic index in the
treatment of syphilis
● Diagnostic for Syphilis if reactive in dilution
1:16 or higher
c. CSF VDRL
● Boerner agglutination slide which has concave
wells 16mm in diameter and 1.75mm deep is
used
● The serum is not heated before testing
● The antigen must be diluted before use with
10% saline solution
● Antigen delivery: 21 or 22g (1/100 mL)
ROTATION:
VDRL serum: 180 rpm for 4 mins
VDRL CSF: 180 rpm for 8 mins
Serological Tests
A. Non-Treponemal Test: RPR
Rapid Plasma Reagin (RPR)
● Modified VDRL test
● More sensitive but less specific than VDRL
● Any positive result must be followed by a
VDRL or other more specific procedures
Antigen is similar to VDRL with the addition of the
following:
a. Charcoal
● Makes test easier to read
b. EDTA
● Prevents oxidation of lipids
c. Thimerosal
● Preservative
d. Choline Chloride
● Inactivate the complement
Serological Tests
 IMMUNOLOGY AND SEROLOGY
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● Patient serum is heat inactivated and made

with a sorbent consisting of non-pathogenic
A. Non-Treponemal Test:
Treponemes (Reiter strain)
RPR
● Nichol’s strain of T.pallidum have been fixed to
slides used for the test

B. Treponemal tests
● Specific treponemal antigens
● Used to detect Treponemal antibodies Fluorescent Treponemal Antibody Absorption Test
developed in response to Treponemal (FTA-ABS)
infections
● Highly specific and sensitive
● Used as verification procedures
a. Agglutination tests:
● Treponema pallidum Agglutination tests (TPA)
● T.pallidum Hemagglutination (TPHA)
● Microhemagglutination- Treponema pallidum
(MHA-TP)
● Hemagglutination Treponemal Test for Syphilis
(HATTS)

b. Immunofluorescence
● Fluorescent Treponemal Antibody Absorption
(FTA-ABS) B. True Treponemal tests
Particle Agglutination (PA) tests
c. Complement fixation tests ● Originally used sheep RBCs coated with T
d. Immobilization tests (TPI) pallidum antigen
● Serodia T pallidum particle agglutination
B. Treponemal tests (TP-PA) test
Fluorescent Treponemal Antibody A
● Indirect fluorescent antibody test
● Test of choice to run with with darkfield
microscopy
● Earliest serological test to be positive

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○ “Doubtful result”: range of 20-50%

Testing Algorithms
● The traditional testing algorithm for syphilis
involves screening the sample with a
nontreponemal test and confirming any
positive results with a more specific
treponemal test

GROUP A STREPTOCOCCAL INFECTION

● Caused by Streptococcus pyognes
● major sites of infection are the upper
respiratory tract and skin with pharyngitis and
empetigo being the most common
● May lead to rheumatic fever and
post-streptococcal glomerulonephritis
FIGURE 21–5 TP-PA test results. Row A: positive control.
Row B: negative control. Row C: serum from a positive
patient. T pallidumsensitized gel particles were placed in
column 3 of each row and unsensitized gel particles were
pipetted into column 4 of each row. Positive wells (A3 and
C3) are indicated by a diffuse mat of particles that spread
over the surface of the well, whereas negative results are
indicated by a compact button. (Linda Miller.)

C. True Treponemal tests

Immobilization tests (TPI)
● Considered as standard or reference to which
all other treponemal tests are evaluated
● Involve mixing of patient serum with live,
actively motile T.pallidum extracted from
testicular chancre of a rabbit and complement
● Test is considered positive if >50%

● Treponemes are immobilized

○ Positive: If 50% or more Treponemes
are immobilized VIRULENCE FACTORS
○ Negative: If fewer that 20% are ● The M protein is the major virulence factor
immobilized
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● Immunity to Group A streptococci (GAS)
appears to be associated with antibodies to
the M protein
● Additional virulence factors include various
exotoxins that may be produced during the
course of a infection
● Additional extracellular substances include the
enzymes streptolysin O, deoxyribonuclease B
(DNase B), hyaluronidase, nicotinamide
adenine dinucleotide (NAD), and
streptokinase
Diagnostic Tests for Streptococcal Infection
CULTURE
● Appear as small translucent colonies
surrounded by a clear zone of β hemolysis
● Identification is made on the basis of
susceptibility to bacitracin, testing for
L-pyrrolidonyl-β-naphthylamide (PYR) activity,
or through Lancefield typing
FIGURE 20–10 Throat culture plate showing a positive result
for beta hemolytic Group A streptococci (S pyogenes).
Bacterial colonies not producing beta hemolysis represent
indigenous microbiota of the oropharynx. (James Vossler.)
Detection of Group A Streptococcal Antigens
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A. Streptolysin O antigen
● Oxygen-labile hemolysin active against human
and rabbit RBC
● Its reduced form, capable of lyzing human or
rabbit rbc
● It unites with an antibody (ASO) and thereby
loses its lytic activity
• DNAse B
• Hyaluronidase
• Streptokinase
● Lateral flow immunochromatographic assays
(LFA) are increasingly being used for the
detection of bacterial, viral, fungal, and
parasitic antigens in clinical samples
The most diagnostically important antibodies are the
following: Anti-streptolysin O (ASO), anti-DNase B,
anti-NADase, and anti-hyaluronidase (AHase).
1. Anti-Streptolysin O titer
● Based on neutralization of the hemolytic
activity of Streptolysin O
● Normal: Titer of 166 Todd units or below
● Moderately elevated: if the titer is atleast 240
Todd units in an adult and 320 in child
● ASO increases within 1-2 weeks after infection
and peak between 3 to 6 weeks following
initial symptoms
Anti-Streptolysin titer
Traditional Method
● Involves dilution of the unknown serum to
which a measured amount of Streptolysin O
antigen is added
● 5% suspension rabbit or human RBC are
added as indicator
● The tube containing the least amount of
serum (one with highest dilution) which
demonstrate no hemolysis is the ASO TITER
[use of 14 tubes → tube 13 does not contain
ASO control]
● Test tube 13: RBC Control no hemolysis
→ no hemolysis
● Test tube 14: ASO Control complete
hemolysis → complete hemolysis
Detection of Streptococcal Antibodies
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2. Anti-Dnase B Testing
● Sometimes appear earlier than ASO in
streptococcal pharyngitis
● Measurement is based on neutralization
● If anti-DNAse B antibodies are present they
will neutralize the reagent DNAse B
● Presence of DNAse is measured by its effect
on DNA-methyl green conjugate.
● The conjugate is in its intact form, but when
hydrolyzed by DNAse, the methyl green is
reduced and becomes colorless
● Tubes are graded for color:
○ 4+ = intensity of color is unchanged
[positive result]
○ 0 = total loss of color [negative
result]
3. Streptozyme testing
● Slide agglutination screening test for
detection of antibodies to several
streptococcal antigens
● Sheep RBC’s are coated with streptolysin,
streptokinase, hyaluronidase, DNAse and
NADase so that antibodies to any of the
streptococcal antigens can be detected

Serology and Molecular Detection of Viral Infections

Immune Defenses Against Viral Infections

● Viruses are obligate intracellular pathogens
that rely on the host cell for their replication
and survival
● Innate immunity provides the first line of
protection against viral pathogens
● Type I interferons and natural killer (NK) cells

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● Virus-specific antibodies are produced by B

cells and plasma cells
Viral Escape Mechanisms
● Cell mediated Immunity
● Frequent genetic mutations and Antigenic
variation
● Escape the action of components of the
innate immune system
● Suppresses the adaptive immune system

Hepatitis Type/ Family Transmission

Viruses

Hepatitis A Picornavirus Fecal-Oral

(HAV)

Hepatits B Hepadnavirus Parenteral

(HBV) (needle),
sexual,
perinatal
(infected
mother to
infant)

Hepatitis C Flavivirus Parentera

(HCV)

Hepatitis D Parentera
(Delta Virus) (or similar to
Hepatitis B)

Hepatitis E Calicivirus Fecal-oral

(HEV)

Hepatitis A
● Known as infectious hepatitis
● Short incubation hepatitis (ave of 28 days)
HAV Antigens:
1. HAV antigens
● Shed in feces of infected individual
HAV Antibodies:
1. IgM Anti- HAV
● Marker of acute hepatitis A
● Peak during 1st month of illness and decline
within 6-12 months
● Routinely detected by solid-phase antibody
capture ELISA

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2. IgG Anti-HAV 4. Anti-HBs

● Produced as a result of natural infection or ● Appears during the recovery period of acute
immunization hepatitis B, weeks to months after HBsAg
● Indicate immunity to HAV produced
● Detected by competitive inhibition ELISA Tests ● Persists for years and provide protective
immunity
Hepatitis B
● Also produced after immunization with
● Serum Hepatitis
hepatitis B vaccine
● Dane particle: complete HBV that causes
infection Serological markers in acute hepatitis
HBV antigens:
1. Hepatitis B surface antigen (HBsAg)
● First marker to appear
● Indicator of active infection or chronic
infection
● Important marker in screening blood donors
2. Hepatitis B Envelope Antigen (HBeAg)
● Present during periods of active replication of
the virus FIGURE 23–4 Typical serological markers in acute hepatitis
● Indicates a high degree of infectivity when B. Solid lines represent viral antigen concentrations,
present whereas dashed lines indicate antibody concentrations.
● High vertical transmission Each antigen shares the same color with its associated
antibody.
3. Hepatitis B Core Antigen (HBcAg)
● Not detectable in serum because of the viral
envelope that masks it
● Detected only through biopsy of the infected
liver
● Not considered as a serologic marker
HBV antibodies:
1. IgM Anti-HBc
● 1st antibody to be produced FIGURE 23–5 Typical serological markers in chronic hepatitis
● Indicator of current or recent infection B.
● Useful in detecting infection during the “core
window” period
2. IgG Anti-HBc
● Life-long marker of HBV infection
3. Anti-Hbe
● Indicates that the patient is recovering from
HBV infection
● Marker of convalescence and favorable
prognosis

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Hepatitis C
● Previously classified as “non A –non B”
hepatitis
● Major cause of post-transfusion hepatitis
● Hepatitis C is transmitted mainly by exposure
to contaminated blood, with intravenous drug
use being the main source of infection
● HCV has an average incubation period of 7
weeks
● Majority of infections are asymptomatic
Hepatitis D
● Unclassified, single-stranded RNA virus
● Incomplete virus
● Parenterally transmitted infection that can
only occur in the presence of hepatitis B
● Infection with two virus occur either:
○ Simultaneously as → Coinfection
○ Sequentially → Superinfection in
chronic HBV carriers
● Co-infection: Where a person who is
susceptible to HBV is exposed to someone
who is co-infected with HBV and delta virus,
this results in acute co-infection with both the
viruses at the same time.
○ positive for HDV antibodies and IgM
anti-HBc
● Super-infection: When an HBV carrier is
exposed to infected blood from co-infected
patients then the exposure results in
super-infection of the existing HBV infection
with delta virus; this may result in
development of acute hepatitis (due to delta
virus) in an HBV chronic carrier.
○ Positive for IgG anti-HBc
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Hepatitis E
● Usually presents as an acute, self-limiting
hepatitis without progression to a chronic
carrier state
● Associated with a high rate of mortality in
pregnant women
“water-borne hepatitis”
HEV Antibodies
● IgM anti-HEV is typically present during the
acute infection but rapidly declines in the
early recovery period
● ELISA, Western blot and fluorescent antibody
blocking assay
HEV RNA
● Detected in feces of most patients for about 2
weeks after the onset of illness, but may
persist longer in some cases
● Identified by means of PCR
HUMAN IMMUNODEFICIENCY VIRUS
● Etiologic agent of the Acquired
Immunodeficiency Syndrome (AIDS)
HIV-1
● Formerly called human T cell lmphotropic
virus-type III (HTLVIII),
Lymphadenopathy-associated Virus (LAV), and
AIDSassociated retrovirus (ARV)
● Causes AIDS in US, Europe
HIV-2
● Endemic in West Africa
● Less pathogenic and has a lower rate of
transmission
Main Structural Genes:
1. Env (envelope)
● gp120, gp41 (markers; proteins involve in
attachment of cd4 to HIV)
● Attachment and fusion to CD4 cells
2. Gag (group antigen) gene
● p55, p15, p17, p24
 IMMUNOLOGY AND SEROLOGY
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3. Pol (polymerase)
a. Reverse transcriptase
b. Integrase
c. RNAse
d. Protease
Laboratory testing for HIV Infection
A. SCREENING
1. ELISA – sandwich ELISA principle (positive result →
bound for confirmatory tests)
• Agglutination Tests
• Dot-Blot Testing
2. Agglutination Tests
- Western Blot Testing (Standard confirmatory
test for number years but replaced by humor
methods) Heterophile Antibodies associated with Infectious
Mononucleosis (IM)
3. Dot-Blot Testing
● Heterophile antibodies are antibodies capable
B. CONFIRMATORY of reacting with similar antigens from two or
more unrelated species
1. Western Blot Testing
Infectious Mononucleosis
WESTERN BLOT
● Caused by Epstein Barr virus
● Target: B cells (CD21)
● It is based on the recognition of the major HIV ● Atypical lymphocyte: T cells reacting to B cells
proteins (p24, gp41, gp120/160) by infected with EBV
fractionating them according to their weight
by electrophoresis and then visualizing their Disease Monitoring
binding with specific antibodies over Two laboratory markers:
nitrocellulose sheets.
● CD4 T-cell count - → prevent infection in
progression to AIDS; <200/uL → stage 3 HIV
infection, <200-499/uL → stage 2, <500/uL
● A positive result: presence of any two of the ● →1
following bands—p24, gp41, and gp120/160 ● HIV-1 RNA level, or “viral load,” - (based on
amplification methods that increase number
of RNAs),” → response to ART (anti-retroviral
● If the test is positive for bands gp41 and/or treatment); successful therapy can result to
p24 in conjunction with a positive EIA test 62undetectable viral load with the help of ART
result, it is regarded as a confirmatory test.

● A negative result demonstrates the absence of

bands

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Heterophile Antibodies associated with Infectious Agglutination with GPK and no agglutination with
Mononucleosis (IM) beef = infectious mononucleosis
● Heterophile antibodies are antibodies capable
3. Monospot test
of reacting with similar antigens from two or
more unrelated species ● based on the agglutination of horse
erythrocytes by heterophile antibody present
Infectious Mononucleosis
in infectious mononucleosis.
● Caused by Epstein Barr virus ●
● Target: B cells (CD21) ● Horse red blood cells (RBCs) exhibit antigens
● Atypical lymphocyte: T cells reacting to B cells directed against both Forssman and infectious
infected with EBV mononucleosis antibodies, a differential
absorption of the patient’s serum is necessary
Laboratory Diagnosis
to distinguish the specific heterophile
1. Paul- Bunnell test
antibody from those of the Forssman type.
● Detect heterophile antibodies in patient
serum when mixed with antigen-bearing
sheep erythrocytes.
● Dilutions of inactivated patient serum are
mixed with sheep erythrocytes, incubated,
centrifuged, and macroscopically examined
for agglutination.
● Positive reactions are preliminary
1. Davidson Differentialtest
● Distinguishes between the heterophile
antibodies that agglutinate the
antigen-bearing erythrocytes of sheep
● Performed only if the preliminary Paul-Bunnell
test is positive in a titer of 1:56 or greater
Laboratory Diagnosis

ADS WITH GPK CELLS ADS. WITH BEEF/OX

ERYTHROCYTES

Antibody to (+) adsorption (-) not adsorbed

Forsmann

Antibody to Serum (+) adsorption (+) adsorption

Sickness

Antibody to IM (-) not adsorbed (+) adsorption

AGGLUTINATION
WITH SHEEP RBC

Antibody to NO AGGLUTINATION AGGLUTINATION

Forsmann

Antibody to Serum NO AGGLUTINATION NO AGGLUTINATION

Sickness

Antibody to IM AGGLUTINATION NO AGGLUTINATION

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