HISTORY AND MILESTONES IN IMMUNOLOGY action of lysozymes
• James Gowan (1924-) - discovered that adaptive immunity is mediated
• Voltaire (1773) - Reported on Chinese practice of variolation.
• Edward Jenner (1798)- smallpox vaccinations - CROSS IMMUNITY
• Haeckel (1862)- phagocytosis
• LOUIS Pasteur (1880-1881) - FATHER OF IMMUNOLOGY
→ live, attenuated chicken cholera and anthrax vaccines; (1885)-
live attenuated vaccine for rabies.
• Elie Metchnikoff (1883-1905) cellular theory of immunity through
phagocytosis
→ NOBEL PRIZE CHAMPION OF CELLULAR IMMUNITY
• Emil Von Behring, Kitasato (1890)- Humoral theory of immunity
proposed
→ CHAMPIONS OF HUMORAL IMMUNITY -- DISSOLVED
SUBSTANCES— ANTIBODIES, ETC.
→ ANTITOXINS --- SERUM OF ANIMALS—HORSES, COWS
→ PASSIVE IMMUNITY ACTIVE IMMUNITY
→ Clostridium tetani--- tetanospasmin – lock jaw syndrome--
neurotoxin
• Robert Koch (1891)- Demonstration of cutaneous hypersensitivity
(REACTION TO TUBERCULOSIS) GRANULOMA
→ VACCINE AGAINST TB.
• Gruber and Durham (1896)- discover agglutination reactions
• Ferdinand Widal (1896)
→ Devised an agglutination reaction for the diagnosis of typhoid
fever. Paul Ehrlich (1900)- Antibody formation theory
• Karl Landsteiner (1900)- discovery of the ABO Blood Group
• P. Portier and Charles Richet (1902)- Immediate hypersensitivity
anaphylaxis - MOST SEVERE FORM OF ALLERGY
• Almoth Wright , Stephen Douglas, Joseph Denys (1903)
→ Discovered opsonins and describe its relation to
phagocytosis
→ NOBEL PRIZE AWARDEES
→ OPSONINS- SUBSTANCES THAT COATS MICROBES TO
MAKE THEM MORE PRONE TO PHAGOCYTOSIS
→ OPSONIZATION
• Von Pirquet and Schick (1906)- describe the relationship of immunity
and hypersensitivity Albert Calmette and Camille Guerin (1921)- 1st
successful vaccine against tuberculosis. - BCG
• Pfeiffer (1894-1895), Buchner- complement mediated cytolysis
• Jules Bordet (1920)- received Nobel Prize for his pioneering works
on complement
→ COMPLEMENT- PRINCIPAL SOLUBLE MEDIATOR OF
INFLAMMATION
• JHONAS Salk, ALBERT Sabin- development of polio vaccine
• George Snell (1903-)- worked out the genetics of the murine major
histocompatibility complex (MHC) and generated the congenic
strains needed for its biologic analysis.
→ PORCINE
→ BOVINE
• Niel Jerne (1911-)- developed the hemolyic plaque assay and
several important immunological theories including an early version of
clonal selection
• Peter Medawar( 1915-1987)- made studies on acquired
immunologic tolerance.
• Jean Dausset (1906-)- an early pioneer in the study of MHC or HLA.
• Baruj Benacerraf (1920-)- discovered immune response genes and
collaborated in the first demonstration of MHC restriction.
• MHC ---- SET OF GENES. FOUND CHROMOSOME # 6
• GENES THAT EXPRESS THE PRODUCTION OF HLA
→ HUMAN LEUKOCYTE ANTIGEN. ----MISNOMER
o FOUND OF ALL NUCLEATED CELLS
o ORGAN TRANSPLANTATION
o ORGAN REJECTION
• 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 of
peptide hormones
→ NOBEL PRIZE AWARDEE
• Alexander Flemming (1922)- described the
by lymphocytes
• Cesar Milstein(1927-) and Georges Kohler (1946)
→ Developed the technique of monoclona antibody formation.
→ NOBEL PRIZE AWARDEE
• 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 Kendall in conducting
quantitative precipitin assay studies on antigen antibody
reactions.
• Max Theiler (late 1930)- 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…
ANTIBODY DIVERSITY; TCR (T-CELL RECEPTOR)
DIVERSITY
• Alic Isaacs and Jean Lindemann (1957)- discovered interferon
• INTERFERON- SUBSTANCES WITH ANTI-VIRAL PROPERTIES
→ BLOCK THE TRANSLATION OF VIRAL PROTEINS
→ 1. ALPHA INTERFERON PRODUCED BY LEUKOCYTES
TYPE 1 IFN
o STIMULATE PRODUCTION OF NK CELLS
→ 2. BETA INTERFERON- PRODUCED BY FIBROBLAST
→ 3. GAMMA INTERFERON- PRODUCED BY
T-CYTOTOXIC CELLS TYPE 2 IFN
ENHANCE FUNCTION OF NK CELL
• Luc Montaigner (1980)
→ isolated a 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
• Mosmann (1986)- Th1 versus Th2 model of T-helper function
• E.Dounal Thomas and Joseph Murray (1991)
→ Introduced the concepts of Transplantation
• 1996-1998- identification of Toll-Like Receptors (TLRS)
→ INVOLVED IN DIRECT PHAGOCYTOSIS
• Peter Doherty and Rolf Zinkernagel (1996)- work on the mechanism
of cellular immune response mediated by T-cells towards virally
infected cells.
→ NOBEL PRIZE AWARDEES ----- PRINCIPLE OF DOUBLE
RECOGNITION ON HOW T LYMPHOCYTES RESPOND
AGAINST INFECTED AND CANCER CELLS.
→ B-LYMPHOCYTES--HUMORAL IMMUNITY-- MAJOR
PRODUCER OF ANTIBODIES
→ T-LYMPHOCYTES—CELLULAR IMMUNITY
o 1. THERE MUST BE ANTIGEN
PROCESSING AND PRESENTATION
BY APCs (ANTIGEN PRESETING
CELL)
o 2. THERE MUST BE ASSOCIATION WITH MHC CLASS 1
OR CLASS 2 MOLECULES
• 2001- discovery of FOX3p, the gene directing regulatory -T-cell
development
• IAN Frazer (2005)- discovery of the Human HPV vaccine
→ HUMAN PAPILLOMA VIRUS--- WARTS
→ RISK FACTOR FOR CERVICAL CANCER
 IMMUNITY- All those physiological mechanisms that endow the animal 5. Nature
with the capacity to recognize materials as foreign to itself and to of Extracellular Antigen Intracellular
neutralize, eliminate or metabolize them with or without injury to its own Infecting organism
tissues Agent
→ Complex reaction involving many different cells, molecules and Chronic,
genes aimed essentially in maintaining the genetic integrity of an granulomatous
individual, protecting it from invasion of substances that can bear infection,
the imprint of a foreign genetic code. 6. Type of Acute pyogenic infection neoplasm,
→ The body’s ability to resist foreign organisms and toxins (poisons) Infection fungal,
that damage tissues & organs parasitic
→ It is a part of a complex system of defense reactions of the body diseases
which can be classified as either innate or acquired.
Contact
TYPES OF IMMUNITY: sensitivity, DTH,
A. NATURAL/INNATE/NON-SPECIFIC/NON-ADAPTIVE IMMUNITY Allograft
- ability of an individual to resist infections by means of normally present Ab-mediated hypersensitivities rejection, GVH
body functions. 7. Variants Autoimmunity response,
B. ACQUIRED/ADAPTIVE/SPECIFICIMMUNITY elimination of
- a reaction resulting from the invasion of a foreign substance. tumors,
formation of
chronic
HALLMARK FEATURES
granulomas
INNATE IMMUNITY ADAPTIVE IMMUNITY
NATURAL IMMUNITY
1. Mechanisms involved are 1. Reinforcement Components:
non-specific 1. External Defense Mechanisms
2. Mechanisms that pre-exist 2. Inducibility - composed of structural barriers that prevent most infectious agents
the invasion of foreign from entering the body.
agents A. Structural/Physical Barriers
3. Components are pre- 3. Specificity a. Intact Skin--LACTIC ACID --SWEAT --pH 5.6
formed b. Mucous membranes of the respiratory and GI tract, GU TRACT---
4. They are non-adaptive, has 4. Diversity- MUCUS
a standardized magnitude of c. Ciliated Epithelium- CILIA
response. d. Lacrimal apparatus---- TEARS ----- LYSOZYMES-
DESTROY GRAM POSITIVE CELL WALL
5. Lacks immunologic 5. Memory - ABILITY TO RECALL e. Sweat, sebaceous glands- SEBUM --- FATTY ACIDS
memory PREVIOUS EXPOSURE WITH THE B. Mechanical Barriers
SAME ANTIGEN/MICROORGANISM a. Peristaltic movement of intestine BOTULIN (Clostridium
6. Specialization botulinum) canned food poisoning
7. Self-Limitation b. Shedding of cells
8. Discrimination c. Coughing and sneezing
d. Flushing action of urine
C. Chemical Barriers
2 TYPES OF ADAPTIVE IMMUNITY
a. Acid pH- lactic acids and fatty acids, HCl pH 1 or 2,
ACTIVE-PASSIVE
Lactobacillus acidophilus in intestine and vagina
1. .NATURAL ACTIVE 1. NATURAL PASSIVE NORMAL FLORA
2. ARTIFICIAL ACTIVE 2. ARTIFICIAL PASSIVE COMPETITIVE EXCLUSION
PATHOGEN AND OPPORTUNISTIC
b. Lysozyme- attacks cell wall of microorganisms and
2 ARMS OF ADAPTIVE IMMUNITY rendered it osmotically sensitive.
HUMORAL (HI) CELL c. Lactoferrin- in human milk
MEDIATED
(CMI) 2. Internal Defense Mechanisms
1. Antibody mediated Cell mediated A. Physiologic Factors
Mechanism a. Body temperature- 36.5 TO 37.5 DEGREES CELSIUS…
2. Cell type B-lymphocytes T-lymphocytes b. Oxygen tension
c. Hormonal balance
B. Basic polypeptides
Direct cell-to-cell a. Spermin- pH dependent polyamine found in semen; inhibits the
3. Mode of Antibodies in biologic fluid contact (APC growth of gram positive bacteria
action AND MHC) or b. Defensin- present in human neutrophil, are catioinic proteins that
soluble products kill microbes by interacting with microbial membrane to form
secreted by cells channels through which important metabolites escapes.
C. Interferons- group of molecules that limit the spread of viral
Defense infections by blocking translation of viral proteins.
Primary defense against bacterial against viral Types:
4. Function infections and fungal a. Alpha IFN
infections, b. Beta IFN
intracellular c. Gamma IFN
organisms, D. Complement- principal soluble mediator of inflammatory response.
tumor antigens, E. Acute phase proteins/REACTANTS- serum molecules in which
and graft concentrations increase rapidly at the onset of inflammatory
rejection, disease
parasites a. CRP
-MOST COMMONLY REQUESTED ACUTE PHASE PROTEIN IN THE
 LAB. TO DETERMINE IF THERE IS INFLAMMATION.
-SENSITIVE BUT NOT SPECIFIC MARKER OF INFLAMMATION.
-C- REACTIVE PROTEIN
-PRIMITIVE ANTIBODY AGAINST C POLYSACCHARIDE OF
PNEUMOCOCCUS (S. pneumoniae)
- 0.5 MG/DL
-UP TO 1000X
-MALES: UP TO 1.5 MG/L
-FEMALES: UP TO 2.5 MG/L
CLINICAL USES:
-MARKER OF ACUTE INFLAMMATION
-MONITOR ANTIBIOTIC/CHEMO-THERAPY
2 MG/L
CRP AGGLUTINATION TEST- POSTIVE OR NEGATIVE, ESTIMATE
OF THE VALUE OF CRP
hsCRP (QUANTITATIVE)----- HIGH-SENSITIVE CRP-- -EARLY
PREDICTOR OF MYOCARDIAL INFARCTION (MI) AND • Granules contain ACP, ALP,and protease
CEREBROVASCULAR ACCIDENT (CVA)
0.1 MG/L 45.92MG /L
b. Serum amyloid A- REMOVES CHOLESTEROL
c. Mannose Binding Protein/LECTIN- BINDS WITH MANNOSE-
OPSONIN, ACTIVATE THE COMPLEMENT SYSTEM
d. Alpha-1-antitrypsin—GENERAL INHIBITOR OF PROTEASE
-NEUTRALIZE ELASTASE (ONE OF DIGESTIVE ENZYMES FOUND IN F. Macrophages
THE GRANULES OF NEUTROPHILS
ELASTASE ---- DEGRADE ELASTIN AND COLLAGEN
CHRONIC SMOKERS-EMPHYSEMA, PULMONARY IDIOPATHIC
FIBROSIS
e. Haptoglobin ---- BINDS HEMOGLOBIN RELEASED
DURING INTRAVASCULAR HEMOLYSIS
HEMOGLOBIN-HAPTOGLOBIN COMPLEX ---------- LIVER
f. Fibrinogen --- PRIMARY AND THE MOST ABUNDANT
COAGULATION FACTOR/PRO-CLOTTING FACTOR
-----RESPONSIBLE FOR THE FORMATION OF CLOT
ESR --- ERYTHROCYTE SEDIMENTATION RATE
g. Ceruloplasmin- BINDS AND TRANSPORT COPPER;
FERROXIDASE (FERROUS TO FERRIC)
-TRANSPORT COPPER TO CYTOCROME B OXIDASE---
AEROBIC ENERGY PRODUCTION—PRODUCE OXYGEN
METABOLITES SUCH AS SUPEROXIDE ANION,
HYRDROXYL RADICAL AND HYPOCHLORITE IONS,
HYDROGEN PEROXIDE
h. Alpha-1-acid glycoprotein
i. Endogenous pyrogens: IL-1- CYTOKINE
PRODUCED BY MONOCYTES
F. Cellular Defense Mechanisms
A. Neutrophils
• 50-70% of circulating WBCs
• Primary granules contain myeloperoxidase, elastase,
proteinase 3, lysozyme, cathepsin G, defensins
• Secondary granules contain collagenase, lysozyme,
lactoferrin, plasminogen activators, ALP, NADPH
• Tertiary granules contain gelatinase and plasminogen activator
• capable of the process Diapedesis (process when
phagocytes move across the blood vessel walls)
• CHEMOTAXIS- phagocytes move toward the site of
infection or inflammation
• CHEMOTAXINS- major phagocyte of the body. Active against
bacterial infections
• Frustrated phagocytosis
• Kamikaze cell/ harikiri cell
• Neutrophilia
B. Eosinophils
• 1-3% of circulating WBCs
• -increases in allergic reactions and parasitic diseases
• -reddish-orange granules
o Primary granules contain ACP, arylsulfatase
o Secondary granules contain Major Basic Protein, eosinophil
cationic protein, eosinophil peroxidase, eosinophil- derived
neurotoxin, phospholipase, histaminase, aminopeptidase and
ribonuclease.
C. Basophils
• Less than 1% of circulating WBCs
• Bluish-purple granules contain histamine, eosinophil
chemotactic factor of anaphylaxis, and heparin.
• involved in immediate hypersensitivity reactions
ANAPHYLAXIS
D. Mast cells SKIN, RESP, GI, GU TRACT
• Can be found on connective tissues.
E. Monocytes
• largest WBC, constitutes 4-10% of circulating WBCs
• possess grayish-blue cytoplasm and is ground-glass in appearance
granules contain peroxidase, ACP, arylsulfatase
• other type of granule contains B-glucuronidase, lysozyme and lipase
• larger version of monocytes on tissues.
• there is increase in the number of ER, lysosomes and mitochondria
• granules contain no peroxidase at all compared with monocytes.
• Monocyte-macrophage system functions in microbial
killing, tumoricidal activity, killing of intracellular
parasites, phagocytosis, secretion of cell mediators and
antigen presentation. (APC)
• MAJOR PHAGOCYTE OF THE BODY
• ACTIVE AGAINST INTRACELLULAR ORGANISMS
FACILITATED/SUSTAINED PHAGOCYTOSIS ---- ASSSITED BY
OPSONINS
o LUNGS: ALVEOLAR MACROPHAGES
o LIVER: KUPFFER CELLS
o BRAIN: MICROGLIAL CELL
o KIDNEYS: MESANGIAL CELLS
o CONNECTIVE TISSUE: HISTIOCYTES
o BONES: OSTEOCLAST
G. Dendritic cells
• function is to phagocytosed antigen and present it to T-helper cells
• Most potent phagocytic cell in the tissues.
• MAJOR APC OF THE BODY
• SKIN AND MUCOUS MEMBRANES: LANGERHAN CELLS
• VITAL ORGANS: INTERSTITIAL DENDRITIC CELL
• LYMPHOID ORGANS: INTERDIGITATING DENDRITIC CELL
PHAGOCYTOSIS
- Engulfment of cells and particulate matter by leukocyte, macrophage
and other cells.
Steps:
1. Physical contact between the WBC and the foreign particle
2. Formation of phagosome
3. Fusion with lysosome
4. Digestion and release of debris to the outside
PATHWAYS OF KILLING PATHOGENS BY PHAGOCYTES
 →DIAPEDESIS
1. Oxygen Dependent →CHEMOTAXIS
Respiratory Burst- occurs when the cytoplasmic pseudopods enclosed the →PHAGOCYTOSIS
particle within a vacuole →TISSUE REPAIR

TYPES OF INFLAMMATION:
1. ACUTE INFLAMMATION--- MEDIATED BY NEUTROPHILS
2. CHRONIC INFLAMMATION --- MEDIATED BY
MONOCYTES/MACROPHAGES--- LEAD TO PERMANENT TISSUE
DAMAGE

MAJOR EVENTS:
1. Tissue damage cause release of vasoactive and chemotactic factors
that trigger a local increase in blood flow and capillary permeability.
2. Permeable capillaries allow the influx of fluids and cells.
3. Phagocytes migrate to the site of the inflammation.
4. Phagocytes and anti-bacterial exudates destroy pathogen.

CARDINAL SIGNS:
1. Rubor- REDNESS
2. Calor- HEAT
2. Oxygen Independent 3. Tumor- SWELLING
-Production of nitric oxide from oxidation of L-arginine by NO synthase 4. Dolor- PAIN
which is produced by IFN-gamma activated cells. 5. Functio Laesa- LOSS OF FUNCTION

TYPES OF PHAGOCYTOSIS
INDIRECT DIRECT
Via opsonin receptors that Receptors that recognize lipid and
recognize opsonins such as IgG, carbohydrate sequences in
CRP, C3b bound to microorganisms
microorganisms
Via Pattern Recognition

PRRs PAMPs (pathogen associated molecular pattern)

TOLL LIKE RECEPTORS
✓ TLR-2 teichoic acid
✓ TLR-4 lipopolysaccharide
✓ TLR-5 flagellin

INFLAMMATION
• the overall reaction of the body to injury or invasion by an
infectious agent.
• TISSUE INJURY
• VASODILATING AGENT:
✓ Histamine
✓ Kinins
• PROSTAGLANDIN ------ pain chemical
• CYCLOOXYGENASE --------- cause platelet aggregation
• ARACHIDONIC ACID-------→ prostaglandin (enhances the effect of
histamine)
• PAIN KILLER----- aspirin

Leukotrienes
• LIPOOXYGENASE
• ARACHIDONIC ACID ------ → leukotrienes (100x more potent than
histamine)

Process of Inflammation
→VASODILATION- increase in blood flow
→INCREASE IN VASCULAR/CAPILLARY PERMEABILITY
 -Cortex- B-cell area, also contains Primary Follicles which
ACQUIRED/ADAPTIVE IMMUNITY- type of smalll amount of T-cell and follicular dendritic cells and
resistance that is characterized by specificity for individual secondary follicles that contains germinal center
pathogen and the ability to remember a prior exposure -Paracortex- T-cell area
(memory) which results in an increased response upon -Medulla- contains differentiated cells and APCs
repeated exposure
Afferent lymphatic (lymphatic fluid with bacteria/
Lymphocyte (2nd most abundant) pathogen enter) --> cortex/paracortex (either B- or T-cells
*T-lymphocytes and B-lymphocyte will be activated)

2 MAJOR STAGES OF LYMPHOCYTE DEVELOPMENT Axillary Lymph Nodes

1. Antigen Independent Stage of Lymphopoiesis- takes Inguinal Lymph Nodes
place on primary lymphoid organs Cervical Lymph Nodes
2. Antigen Dependent Stage of Lymphopoiesis- takes place
in secondary lymphoid organs LYMPHADENOPATHY/LYMPHADENITIS

THE LYMPHOID SYSTEM Lymphocyte traffic/recirculation- movement of

1. Primary Central Lymphoid Organs- developmental lymphocytes from blood to lymphoid organs and back to
sites, maturation site for LYMPHOCYTES the blood
a. Bone Marrow- main source of hematopoietic stem cells -
MATURATION SITE FOR B-CELLS Site of Entry and Exit of Lymphocytes in the lymph
-center for antigen-independent lymphopoiesis node:
b. Thymus- small, flat bilobed organ found in thorax or 1. Afferent arteriole
chest cavity right below the thyroid gland and overlying the 2. Efferent arteriole
heart- MATURATION SITE FOR T-CELLS
-an endocrine gland (thymosine- facilitate maturation) B. Non-encapsulated organs
-Cortex- where thymocytes (immature T-cells) can be -Mucosal Associated Lymphoid Tissue- small masses of
found (85% of population of T-cells0 lymphoid tissue found in intestinal, genitourinary tract and
-Medulla- where mature T-cells can be found respiratory tract
-Thymic stromal cells- include epithelial cells,
macrophages and dendritic cells BALT (Bronchus Tonsils, adenoids
Associated Lymphoid
2. Secondary/Peripheral Lymphoid Organs- activation Tissue)
sites for LYMPHOCYTES GALT (Gut Associated Peyer's patches,
*Antigen- cause the activation of lymphocyte Lymphoid Tissue) appendix
A. Encapsulated Organs CALT (Cutaneous Intraepidermal
1. Spleen- larhest secondary lymphoid otgan that is found Associated Lymphoid lymphocytes
on the ULQ of the abdomen just below the diapragm Tissue)
2 Main Types of Splenic Tissue:
a.Red Pulp- involved in culling process, pitting and platelet *ACUTE SUPPURATIVE APPENDICITIS --> appendectomy
sequestration *APPENDECTOMY- removal of inflammed appendix
*CULLING- process of removing old/senescent RBCs
*PITTING- process of removing abnormal inclusion bodies CD (Cluster of Differentiation)- membrane proteins that
in the RBC serve as surface marker for certain blood cells; to identify
*1/3 produced platelets are stored in the red pulp of spleen blood cells
*SPLENECTOMY- lead to increased platelet count; prone to
infection with encapsulated bacteria CD Markers for T-cells CD2, CD3, CD4, CD8
b.White Pulp contains lymphoid tissue that is arranged CD Markers for B-cells CD19, CD21, CD23
around arterioles as PALS (Periarteriolar Lymphoid Sheet) CD Markers for NK cells CD16, CD56, CD94
-T-cells are found near the central arteriole
-Naive/Virgin/Unstimulated/Inactivated B-cells are found
on the Primary Follicles Monoclonal Antibodies
-Activated B-cells (Plasma Cell & Memory Cell) are found on Anti-CD2 + CD2 ---> POSITIVE (T-cells)
Secondary Follicles (germinal centers)
-Marginal zone contain macrophages STAGES OF B-CELL DIFFERENTIATION

2. Lymph Nodes- junctional filter of the lymphoid system, *B CELLS- MAJOR PRODUCER OF ANTIBODIES
sizes may range from 1mm to 25 mm
*ANTIBODIES- made up of monomer (Y-shaped molecule; 5. Activated B cells- exhibit CD25 on surface, CD25 in turn
made up of 2 heavy chains and 2 light chains) acts as receptor for IL-2
*HEAVY CHAINS: gamma, alpha, mu, delta, epsilon *CD25 + IL-2--> proliferation of activated lymphocytes
*LIGHT CHAINS: kappa, lambda
6. Plasma Cells- large spherical, ellipsoidal cells that
1. PRO-B CELLS- markers include CD19, CD45R, TdT, RAG- contain abundant cytoplasmic immunoglobulins and little
1 and RAG-2 enzymes to no surface immunoglobulins
*TdT (termical deoxyribonucleotide transferase) *specific stage of the B-cell that will produce antibodies
*RAG (recombinase activating gene) *has cytoplasmic immunoglobulins
*eccentric nucleus
2. PRE-B CELLS- contains pre-B cell receptor that is made *short-lived cell
up of 2 mu heavy chains (in cytoplasm) and a surrogate
light chains whose function is to transmit signals to prevent 7. Memory cells- progeny of antigen stimulated B-cells that
rearrangement of any other heavy chains are capable to responding to antigen with increased speed
and intensity
3. Immature B-cells- recognized by the appearance of *has SURFACE IgG
complete IgM molecules on their surface *long-lived cells
-Specificity of the surface immunoglobulin to be
synthesized can already be predicted or noted IgM- marker of acute/present/active infection
-Surface markers that can be seen include receptors for IgG- marker of chronic/past infections; marker of lifelong
complement components such as C3d, CD21 immunity
-B cells capable of producing antibody to self-antigens are
deleted in the marrow through APOPTOSIS (programmed *Application
cell death). Dengue IgM: + (present)
Dengue IgG: + (past)
4. Mature B cells- exhibit IgD and more IgM molecules on IgG and IgM positive: CONVALESCENT
their surface as well as MHC Class II products STAGE/RECOVERY PHASE OF INFECTION

*CD19- present in all populations of B-cells*

ACQUIRED IMMUNITY
IMMUNE SYSTEM
• Lymphatic system
• Computerized war machine
• “defense department” of the body

CELLS OF THE IMMUNE SYSTEM

1. Myeloid Lineage
2. Lymphoid Lineage

Major Functions of the Immune System:

1. Block harmful agents
2. Seek-out invasive pathogens
3. Isolate and neutralize activity of that antigen

ACQUIRED IMMUNITY
- type of resistance that is characterized by specificity for each
individual pathogen and the ability to remember a prior exposure (MEMORY) , which results in an increased response
upon repeated exposure.

LYMPHOCYTES- the key cell involved in adaptive immunity.

• Represents approximately 20% of the circulating WBCs.
• Has a large rounded nucleus which contains a dense nuclear chromatin

2 MAJOR STAGES OF LYMPHOCYTE DEVELOPMENT

1. Antigen Independent Stage of Lymphopoiesis--- TAKE PLACE ON PRIMARY LYMPHOID ORGANS
2. Antigen Dependent Stage of Lymphopoiesis---- TAKE PLACE IN SECONDARY LYMPHOID ORGANS

HEMATOPOEISIS

THE LYMPHOID SYSTEM

1. Primary/Central Lymphoid Organs- developmental sites; MATURATION SITES FOR LYMPHOCYTES
a. Bone Marrow-main source of hematopoietic stem cells.—MATURATION SITE FOR B-CELLS
• Center for antigen-independent lymphopoiesis.

b. Thymus- small, flat bilobed organ found in thorax or chest cavity right below the thyroid gland and overlying
the heart.—MATURATION SITE FOR T-CELLS
• An endocrine gland—HORMONE--- THYMOSINE
• Cortex- where thymocytes 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.2.

Secondary/Peripheral Lymphoid Organs- activation sites FOR LYMPHOCYTES

ANTIGEN--- CAUSE THE ACTIVATION OF LYMPHOCYTE

A. Encapsulated Organs
1. Spleen-largest secondary lymphoid organ that is found on the ULQ of the abdomen just below the
diaphragm

2 Main Types of Splenic Tissue:

a. Red Pulp- involved in culling process, PITTING AND PLATELET SEQUESTRATION
CULLING- PROCESS OF REMOVING OLD/SENESCENT RBCs
PITTING- PROCESS OF REMOVING ABNORMAL INCLUSION BODIES IN THE RBC
1/3 PRODUCED PLATELETS ----STORED IN SPLEEN
2/3 CIRCULATING PLATELETS
SPLENECTOMY--- LEAD TO INCREASED PLATELET COUNT
---PRONE TO INFECTION WITH ENCAPUSULATED BACTERIA.

b. White pulp-contains lymphoid tissue that is arranged around arterioles as PALS (Periarteriolar
Lymphoid Sheath)
• T cells are found near the central arteriole.
• Naïve B cells/VIRGIN B-CELLS/UNSTIMULATED/INACTIVATED B-CELLS are found on primary follicles
• Activated B cells (PLASMA CELL AND MEMORY CELL) are found on Secondary
Follicles (germinal centers)
• Marginal zone contain macrophages

SITES OF ENTRY OF LYMPHOCYTE/ANTIGEN INTO SPLEEN:

a. Specialized Capillaries on marginal sinus
b. Via trabecular artery

2. Lymph Nodes-junctional filter of the lymphoid system. Sizes may range from 1mm to 25 mm.
• Cortex- B-cell area; also contains primary follicles which small amount of T cell and follicular
dendritic cells and secondary follicles that contains germinal center.
• Paracortex- T-cell area
• Medulla- contains differentiated cells and APCs

AXILLARY LYMPH NODES

INGUINAL LYMPH NODES
CERVICAL LYMPH NODES

LYMPHADENOPATHY/LYMPHADENITIS
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
2. Efferent arteriole

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 addresins and other adhesion molecules: CD 34/CD 102 marker
-
T and B cells have homing receptors: L-selectin or LAM

B. Non-encapsulated organs

• Mucosal Associated Lymphoid Tissue- small masses of lymphoid tissue found in intestinal,
genitourinary tract and respiratory tract.

BALT (BRONCHUS ASSOCIATED LYMPHOID TISSUE)- includes the tonsils, adenoids

GALT (GUT ASSOCIATED LYMPHOID TISSUE)- Peyer’s patches, appendix

ACUTE SUPPURATIVE APPENDICITIS

APPENDECTOMY- process of removing the appendix

CALT (CUTANEOUS ASSOCIATED LYMPHOID TISSUE)- includes intraepidermal lymphocytes

CD – CLUSTER OF DIFFERENTIATION

-MEMBRANE PROTEINS THAT SERVE AS SURFACE MARKER FOR CERTAIN BLOOD

CELLS

CD MARKERS FOR T-CELL: CD2, CD3, CD4, CD8

CD MARKERS FOR B-CELLS; CD19, CD21, CD23

CD MARKERS FOR NK (NATURAL KILLER) CELLS: CD16, CD56, CD94

MONOCLONAL ANTIBODIES

ANTI-CD2 + CD2---→ POSITIVE T-CELLS

ANTI-CD19 + CD19 ----> POSITIVE B LYMPHOCYTE

STAGES OF B-CELL DIFFERENTIATION

B-CELL ---- MAJOR PRODUCER OF ANTIBODIES

1. Pro-B Cells- markers include CD19, CD45R, TdT, RAG-1and RAG-2 enzymes

Tdt--- terminal deoxyribonucleotide transferase

RAG- recombinase activating gene

2. Pre-B cells- contains pre-B cell receptor that is made up of 2 mu heavy chains and a surrogate light chain
whose function is to transmit signals to prevent rearrangement of any other heavy chain genes.

-----mu heavy chains in cytoplasm

ANTIBODY- MADE OF MONOMER AS BASIC UNIT

MONOMER- Y-SHAPED MOLECULE; MADE OF 2 HEAVY CHAINS AND 2 LIGHT CHAINS

IgG, IgA, IgM, IgD, IgE

HEAVY CHAINS: GAMMA, ALPHA, MU, DELTA, EPSILON

LIGHT CHAIN: KAPPA, LAMBDA

3. Immature B-cells- recognized by the appearance of complete IgM molcules on their surface.
• Specificity of the surface immunoglobulin to be synthesized can already be predicted or noted.
• Surface markers that can be seen include receptors for complement components such as C3d, CD21.
• B-cells capable of producing antibody to self-antigens are deleted in the marrow through APOPTOSIS –
programmed cell death

4. Mature B cells- exhibit IgD and more IgM on their surface as well as MHC Class II products
5. Activated B cells- exhibit CD25 on surface. CD25 in turn acts as receptor for IL-2

CD25 + IL-2 --------→ proliferation of activated lymphocytes

6. Plasma Cells-large spherical, ellipsoidal cells that contain abundant cytoplasmic immunoglobulins and liitle to no
surface immunoglobulins.--- MAJOR PRODUCER OF ANTIBODIES

PLASMA CELL- PRODUCES ANTIBODIES

-HAS CYTOPLASMIC IMMUNOGLOBULINS/ANTIBODIES
-ECCENTRIC NUCLEUS
- SHORT-LIVED CELL.

7. Memory B Cells- progeny of antigen stimulated B-cells that are capable of responding to antigen with increased
speed and intensity.

-SURFACE IgG
-long-lived cells

WINDOW PERIOD

RT-PCR---MOLECULAR BIOLOGY

IgM—marker of acute/present/active infection

IgG--- marker of chronic/past infection

----- marker of lifelong immunity

DENGUE IgM: +
DENGUE IgG: +

CONVALESCENT STAGE/RECOVERY PHASE OF INFECTION- IF IgM & IgG are both positive

CD 19- present in all population of B cells.

STAGES OF T CELL DIFFERENTIATION

1. Pro-thymocyte- possess CD44 and TdT

2. Double Negative Thymocyte-lack CD4 and CD8 antigens.
-possess CD2, CD5, CD7, CD45R
3. Double Positive Thymocyte- express both CD4 and CD8 antigens on their surface as well as CD3-αβ (TCR)
-would undergo Positive and Negative Selection
4. Mature T cells- 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+
• 1/3 CD3+
5.. Activated T cells- express receptors for IL-2 and produce cytokines.
• Cytokines functions include assiting B cells in the commencement of antibody production,
killing of tumors and target cells, rejection of grafts, stimulation of hematopoiesis in the
bone marrow and initiation of delayed hypersensitivity allergic reactions.
6. T memory cells
NATURAL KILLER CELLS/jjgb IMS Lecture No.2
-Larger than T and B cells; contains a kidney shaped nucleus with condensed chromatin and prominent
nucleoli.
- Has high cytoplasmic:nuclear ratio
- Cytoplasm has many azurophilic granules.
- Constitutes 5-15% of the circulating lymphoid pool; found mainly in spleen and blood.
- No specific surface markers
- Possess CD16, CD56, CD 94
- Lack CD3, 4 and 8
- Play a complementary role to CD8+ T cells
- Become LAK cells in response to IL-2

MECHANISMS OF NK CELL CYTOTOXICITY

1. It is brought about by the balance between activating and inhibitory signals that enables NK cells to distinguish
healthy cells from infected or cancerous cells.
- Killer cell inhibitory receptors (KIRs)
- NKG2D- binds to MICA and MICB proteins on diseased or cancerous cells
- If an inhibitory signal is not produced, NK cells will release PERFORINS AND GRANZYMES

2. Antibody Mediated Cell Cytotoxicity

- Through binding of IgG-coated cell with CD 16

LABORATORY IDENTIFICATION OF LYMPHOCYTES

1. Density Gradient Centrifugation with Ficoll-Hypaque
2. Flourescence Microscopy
a. Direct Immunofluorescence- use monoclonal antibodies with a fluorescent tag (e.g. fluorescein and
phytoeryhtrin [490 nm]; rhodamine [545 nm].
b. Indirect immunofluorescence- uses unlabeled antibodythat first combines with the antigen by itself and a
second antibody that is complexed with a dye.
3. Cell Flow Cytometry/Fluorescence 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. Fluorescent antibodies are used to screen of
subpopulation of T and B cells.
Components:
a. Sample delivery system
b. A laser for cell illumination
c. Photodetectors for signal detection
d. Computer based management system
4. Rosetting
5. ELISA
T CELLS B CELLS

-Developed in the thymus -Developed in the bone marrow

-Found in blood (60-70% of circulating
lymphocytes), thoracic duct fluids, lymph nodes
Identified by rosette formation with sheep RBC
-End product of activation are cytokines
-Antigens include CD2, 3, 4 and 8
-Located in the paracortical region of lymph
nodes
-Found in the bone marrow, spleen and lymph
nodes
-Identified by surface immunoglobulins
-Antibody
-CD19, 20, 21, 40 and MHC Class II
-Located in the cortical region of lymph nodes
MAJOR HISTOCOMPATIBILITY COMPLEX

• Large multi-gene locus consisting of several thousand kilobase pair of DNA on a single chromosome.
• 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.
• In humans , it is referred to as Human Leukocyte Antigen (HLA) complex located at chromosome # 6.

NATURE OF HLA ANTIGENS:

1. Glycoprotein component of cell membrane
2. Present in all nucleated cells
3. Products of the genes of MHC

• Inheritance of HLA genes – controlled by a MHC located in the short arm of chromosome # 6

• Major regions in the MHC: D, B, C and A

ORGANIZATION BASED ON THEIR: TRAITS UNDER THEIR CONTROL:

BIOCHEMICAL COMPOSITION:

1. Class I- A, B, C, E, F, G, H, X - transplantation antigens, serologically detected membrane antigens,

Cellular target antigens for cell mediated lympholysis

2. Class II- DP, DQ, DR, DM, DN , DO - I Region leukocyte antigen

-T and B cell interaction
- Immune response
- Mixed Leukocyte Reaction
- Graft versus Host reaction
- Tumor virus susceptibility
- Peptide transport
- Generation of peptides from cytosolic proteins

3. Class III- complement components (C4a, C4b, C2), cytochrome p450, - serum protein molecule
21-hydroxylases and TNF (tumor necrosis factor) - complement levels
-Cytokines and enzymes

PROPERTIES OF MHC MOLECULES

CLASS I CLASS II
1. Genetic loci HLA- A, B, and C HLA SB (DP), DC (DQ), DR

2. Chain Structure α chain + β2 microglobulin (C#15) α chain + β chain

3. Cell distribution all nucleated cells APC ( macrophage, dendritic cells,

Langerhan cells, T and B cells, endothelial,
epithelial and stromal cells )

4. Presents antigen to T- cytotoxic cells (CD8+) T- helper cells (CD4+)

5. Source of peptide fragments Proteins made in the cytosol Endocytosed plasma membrane and
extracellular proteins

6. Polymorphic domains α 1 and α 2 α 1 and β 1

• The highly polymorphic genes within the MHC gene segment of DNA code for a wide range of cell surface structures that T cells must
recognize in association with foreign antigen for a successful immune response.

• MHC Restriction or MHC Restricted Recognition- the process whereby the MHC control interactions between cells. It involves
the recognition of foreign antigen in association with Class I or Class II molecules.

Reactions considered as MHC RESTRICTED:

1. Antigen Presentation
2. T and B cell cooperation
3. Cytotoxic T cell interaction with target cells

Application of HLA typing/Matching:

1. Organ Transplantation 4. Studies of Racial Ancestry and Migration
2. Paternity Testing 5. For diagnostic and genetic counselling
3. Forensic Medicine, Anthropology 6. Basic research in immunology

/jjgb IMS Lecture No. 4

HLA DISEASE ASSOCIATION

HLA- B27 ANKYLOSING SPONDYLITIS; REITER’S SYNDROME

HLA- B47 CONGENITAL ADRENAL HYPERPLASIA
HLA- B5 BEHCET’S DISEASE
HLA-CW6 PSORIASIS VULGARIS
HLA- DR3 TYPE 1 DM
HLA- DR4 RHEUMATOID ARTHRITIS
HLA-DR5 CHRONIC LYMPHATIC LEUKEMIA
GOLD-INDUCED NEPHROPATHY
KAPOSI’S SARCOMA

Methods for detecting HLA Antigens: Histocompatibility Testing

A. 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, RFLP- RESTRICTION FRAGMENT LENGTH POLYMORPHISM))

/jjgb IMS Lecture No. 4

START OF MIDTERMS DISCUSSION… ( but continuation of the last topic in prelims)

STAGES OF T CELL DIFFERENTIATION

1. Pro-thymocyte - possess CD44 and TdT (Terminal deoxyribonucleotidase)
2. Double Negative Thymocyte - lack CD4 and CD8 antigens.
-possess CD2, CD5, CD7, CD45R
3. Double Positive Thymocyte - express both CD4 and CD8 antigens on their surface as
well as CD3- αβ (TCR) T- Cell Receptor
-would undergo Positive and Negative Selection
-POSITIVE SELECTION – The double positive Thymocyte that cannot recognize self MHC
will undergo apoptosis.
-NEGATIVE SELECTION- Thymocyte that can destroy self-antigens will undergo
apoptosis.
TCR – is made up of CD3 molecule and aβ chains
-- made of 8 polypeptide chains
-- 6 of the polypeptide chains is present in CD3
-- 1 of the polypeptide chains is present in a chains
-- 1 of the polypeptide chains is present in β chains
CD3 – made up of the constant regions
aβ chains – made up of variable regions-----these variable regions, IT BINDS WITH
ANTIGEN.
-would undergo Positive and Negative Selection

4. Mature T cells - represents those population of thymocytes that had survived

positive and negative selection
- express only 1 of either CD4 or CD8 on their surface.
1. 2/3 CD4+ T-CELL, this is called T-Helper cells/T-inducer Cells --- MAJOR
PRODUCER OF CYTOKINES.
2. 1/3 CD8+ T-CELL, this is called T-cytotoxic cells/T-killer cells/.---- KILL
TARGET CELLS (cancer cells/virally infected cell VIA DIRECT CELL TO
CELL CONTACT)
3. T-SUPPRESSOR CELLS- this is called ,T-regulatory cells – involved in
the prevention of auto-immune disease.
4. GAMMA DELTA T CELL – behave like natural killer cells.
- Does not required MHC
 (these Mature T-cell will now go to secondary lymphoid organs such as spleen, lymph nodes, mouth)
(waiting for their corresponding antigen to come and when this mature T-cell react with corresponding
antigen it will become ACTIVATED T-CELL)

5. Activated T cells- express receptors for IL-2 and produce cytokines.

IL-2 + CD25 = PROLIFERATION OF ACTIVATED T-CELL
- Cytokines functions include assisting B cells in the commencement of antibody production,
killing of tumors and target cells, rejection of grafts, stimulation of hematopoiesis in the bone
marrow and initiation of delayed hypersensitivity allergic reactions.
6. T memory cells

NATURAL KILLER CELLS (3rd type of lymphocyte)

- Larger than T and B cells; contains a kidney shaped nucleus with condensed chromatin and
prominent nucleoli.
- Has high cytoplasmic: nuclear ratio
- Cytoplasm has many azurophilic granules.
- Constitutes 5-15% of the circulating lymphoid pool; found mainly in spleen and blood.
- No specific surface markers
- Possess CD16, CD56, CD 94
- Lack CD3, 4 and 8
- Play a complementary role to CD8+ T cells

- Become LAK cells in response to IL-2

LAK cells – LYMPHOKINE ACTIVATED KILLER CELL
MECHANISMS OF NK CELL CYTOTOXICITY
1. It is brought about by the balance between activating and inhibitory signals that
enables NK cells to distinguish healthy cells from infected or cancerous cells.
 Killer cell inhibitory receptors (KIRs)
 NKG2D receptors - binds to MICA and MICB proteins on diseased or cancerous cells
 If an inhibitory signal is not produced, NK cells will release PERFORINS (will create pores)
AND GRANZYMES (will digest and destroy cancer cell)
ALL HEALTHY CELLS IN THE BODY HAS MHC CLASS I ON THE SURFACE
 - if the cell is healthy and it has IL-1, the Killer cell inhibitory receptors (KIRs) will bind to
IL-1 it creates a signal for NK Cell for the cell not to be destroyed
CANCER CELLS/VIRALLY INFECTED CELL----- the MHC CLASS I DIMINISH
 - All cancer cells produces MICA and MICB proteins (THEY ARE STRESS PROTEINS)

2. Antibody Mediated Cell Cytotoxicity/ Antibody Dependent Cellular Cytotoxicity

(ADCC)
 Through binding of IgG-coated cell with CD 16

LABORATORY IDENTIFICATION OF LYMPHOCYTES

1. Density Gradient Centrifugation with Ficoll-Hypaque
Ficoll-Hypaque- is a reagent with a specific gravity of 1.077 – 1.114
(mix ficoll-hypaque w/patient’s blood then centrifuge, it will separate lymphocyte from other
components of blood and the you can pipette it out)
2. Flourescence Microscopy
- Direct Immunofluorescence - use monoclonal antibodies with a fluorescent tag
(e.g. fluorescein and phytoeryhtrin [490 nm]; rhodamine [545 nm].
- Indirect immunofluorescence - uses unlabeled antibodythat first combines with
the antigen by itself and a second antibody that is complexed with a dye.
MONOCLONAL ANTIBODY:
if you want to count B-Lymphocyte – use anti CD 19, CD21, CD23 + fluorescent dye
if you want to count T-Lymphocyte – use anti CD2, CD3, CD4, CD8 + fluorescent dye
 3. Cell Flow Cytometry/Fluorescence 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 - Fluorescent antibodies are used to screen of
subpopulation of T and B cells.

4. Rosetting
- lymphocyte suspension + Sheep RBC (it has receptor for CD2)
Components:
a. Sample delivery system
b. A laser for cell illumination
c. Photodetectors for signal detection
d. Computer based management system
4. Rosetting
5. ELISA

COMPARISON OF T AND B CELLS

T cells B cells
Developed in the thymus Developed in the bone marrow

Found in blood (60-70% of circulating Found in the bone marrow, spleen and lymph
lymphocytes), thoracic duct fluids, lymph nodes nodes

Identified by rosette formation with sheep RBC Identified by surface immunoglobulins

End product of activation are cytokines Antibody

Antigens include CD2, 3, 4 and 8 CD19, 20, 21, 40 and MHC Class II

Located in the paracortical region of lymph Located in the cortical region of lymph nodes
nodes
ANTIGEN
─ A substance that reacts with antibody of sensitized T cells
but may or not may provoke an immune response.
─ Antigen binds with either antibody or sensitized T cells
─ Synonym of antigen is Immunogen
─ The only difference between the two term is that
Immunogen is the only one can cause formation of
antibody or immunoglobins or formation of sensitized T
cells.
IMMUNOGEN
─ A macromolecule capable of eliciting the formations of
immunoglobins or sensitized cells in an immunocompetent
host.
─ Both antigen and immunogen can bind with antibody or
sensitized T cells, but only immunogen can stimulate the
production of antibodies or caused the activation of T cell.
*ALL IMMUNOGENS ARE ANTIGEN BUT NOT ALL
ANTIGENS ARE IMMUNOGENS.
IMMUNOGENIC
─ A substance that is capable of causing antibody production
FACTORS INFLUENCING THE IMMUNE RESPONSE
 Age - newborns and elderly are immunocompromised
(common of Pneumonia and Respiratory infection)
─ Newborns: immunocompromised because their
immune system is not yet developed
─ Elderly: their immune system is already
diminished
─ Adults (middle age): immune system is mature
and develop and at its full potential
 Dose - dose of antigen can also affect the response, the
immune response against antigen
─ More antigen of microbe enters the body, then
much faster the immune response
 Route of inoculation – intravenous route (blood) is the
fastest for an antigen to activate our immune system or
to cause antibody production
─ Intramuscular, etc. (slower, because it needs to
diffuse first in skin, tissue and wall of blood
vessel before in the blood)
 Health status of the host – when stress its much easier
to get sick, puyat, etc. (drink vit. C, A and zinc)
 Genetics – there are some genes that can predisposed
us to certain diseases. (ex. Black people have resistant
to malaria because of their genes)
CHARACTERISTICS OF AN IMMUNOGEN
1. Foreigness – the more foreign a substance is the
more immunogenic (we only produce antibody
against foreign antigen) B cell are the common
release antibody
2. High molecular weight – it must be big or large
(>10,000 daltons of HMW, if its < cannot stimulate
antibody production)
3. Molecular/chemical complexity - Proteins are the
most immunogenic followed by carbohydrates.
*Proteins, carbohydrates, Glycoproteins, Glycolipids
(it can cause antibody production and most antigen are made up
of those substances)
*(pure)Lipids and (pure) nucleic acid are not immunogenic
(poor immunogenic)
Immunogenic order: Proteins-carbohydrates-glycoproteins-
glycolipids-lipids-nucleic acid
4. Digestability – it must be broken down into small
pieces for T cells
5. Availability
Two main parts of antigen (hapten and carrier)
 Hapten
o Nonimmunogenic materials that when
combined with a carrier create new antigenic
determinants.
o ANTIGENIC DETERMINANT/EPITOPE-
Molecular shape or configurations that are
recognized by antibody or T cells. May be
either linear or conformational or have
repeating units or different specificities.
o Contains the epitope/antigenic determinant.
o Epitope – the receptor of antigen for the
antibody
o the particular part of antigen that binds with
antibody is the Hapten, particularly the
epitope. (Binding site for the antibody)
 Carrier
o Responsible to give the antigen its required
size to be
antibody
The big oblong HAPTEN (small)
is the carrier.
-
○ the antibody specifically reacts with the antigen,
so if the antibody are not specific to the epitope
of antigen it wont bind. Antigen-antibody
reaction is specific, the sequence of amino acids
 of epitope of antigen are match to the sequence OTHER CATEGORIES OF ANTIGENS:
of amino acid of the antibody.
THYMUS DEPENDENT
○ the antibody has Y-shaped, and the y shaped ANTIGEN
there is called Fab and the stem is Fc.
Capable of mounting
immune response provided
there’s assistance from T
cells
Fc
Capable of stimulating B
cells
Capable of inducing
production of memory cells
THYMUS INDEPENDENT
ANTIGEN
Can mount immune response
without the help from T-
helpercells(CD marker:CD-
4)
Capable of stimulating B
cells
Not capable of inducing
proliferation of memory cells

Fab
Can induce B cell to form Only IgM can be produced
○ antibody contains 1 Fc and 2 Fab and the end of different immunoglobin
each Fab have paratope (the binding site of classes (IgGAMED)
antibody for the antigen)
Examples: viral Examples:
VALENCE hemagglutinin, diphtheria Type 1: bacterial
toxin, PPD polysaccharide, Brucella
○ number of combining sites abortus
-has inherent mitogenic
○ It means the number of hapten or epitope are capacity (mitosis-cell
available in the antigen or the number of binding sites division)
Type 2: Pneumococcal
TYPES OF ANTIGEN: polysaccharide, salmonella-
polymerized flagellin, hapten
1. Autoantigens/self-antigen – antigen that belongs to
conjugated fill (polysucrose),
the host. dextran
- Ex. Red cell specific antigens, HLA
(human leukocyte antigen )
2. Alloantigens/isoantigen – derived from the body of
other individuals of the same species
- Ex. Blood from donors
3. Heteroantigens – derived from other species ─ CD-4 is also the receptor of the human cells for HIV. The
- Ex. Covid-19 (virus) main target cell of HIV inside the human body is T-helper
4. Heterophile antigens – those that exist in unrelated cells.
plants or animals, but which are either identical or
closely related in structure so that antibody will cross ─ T-helper cell is the central cell of the immune system.
react with antigen of the other. ADJUVANTS – A substance administered with an immunogen
- Have like structure related to the antigens in that increases immune response.
our body
- Heterophile are heteroantigens but the Ex. Aluminum salts, complete Freund’s adjuvant
antigens from the other species are
─ Commonly mix with vaccines (vaccines contains
somewhat like the antigen present in our
antinaglutide or inantiglutide antigen di ko sure spell
body
pero ganyan pronounce)
- Ex. Polysaccharide type XIV of
─ There’s a vaccine also that contains anti-toxin, anti-
pneumoncoccus reacting with anti-A
tetani, anti-diphtheria but for the vaccine containing
antisera.
attenuated or inactivated antigen are mixed with
(it closely resembles the A-antigen that are found in the red cell
adjuvant, because it increases the size of
of type A individual)
immunogen.
─ Adjuvant coats the attenuated or inactivated antigen
and those increase the size of immunogen.
─ So, when the immunogen is big it wont easily diffuse
to tissue or walls of blood vessels, prolonging its
existence in an area; trap to a tissue. And in that way
when the vaccine is injected to you the antigen can’t
 go anywhere and the antibody B cell can easily bind
to it.
─ When the immunogen of vaccine is trap in a
particular area it increase the number of macrophages
involved in antigen processing
─ Vaccine stimulates T-cell
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
Divided into 3 regions: organization based on their Biochemical
composition
• Class II
• Class III
I. MAJOR HISTOCOMPATIBILITY COMPLEX (MHC)
a) Large multi-gene locus consisting of several thousand
kilobase pair of DNA on a single chromosomes
b) 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
c) In humans, it is referred to as Human Leokocyte
Antigen (HLA) complex located at chromosome #6
• Each human has 26 chromosome
• Chromosome #6
‒ Also called MHC
‒ Its DNA contains MHC genes which involved in
controlling the exchange of tissue (tissue
compatibility)
‒ Major product: Human leukocyte Antigen
(HLA)
• HLA (Human Leaukocyte Antigen)
‒ Match during organ transplantation
‒ HLA type of the recipient must have the same
HLA type to the donor
• MHC genes is involved in the interaction of immune
cells
• MHC molecules are the products produced by MHC
genes
‒ e.g. HLA one of the many product produced by
MHC genes
Chromosome #6
 Expressed the production of MHC class II molecules
 Genes:
− DP, DQ, DR (Major gene)
Gene Products:
− HLA DP
− HLA DQ
− HLA DR
− DM, DN, DO (Minor gene)
 Expressed the production of MHC class III molecules
  Genes: ─ The highly polymorphic genes within the MHC gene
− Complements: C2, C4A, C4B segment of DNA code for a wide range of cell surface
− Cytochromes p450 structures that T cells must be recognize in association
 Can produce enzyme: 21-hydroxylases and TNF with foreign antigen for a successful immune response
(Tumor Necrosis Factor) ─ MHC restriction or MHC restricted recognition
 Process whereby the MHC control
• Class I interations between cells
 Expressed the production of MHC class I  It involves the recognition of foreign
molecules antigen in association with Class I or
 Genes: (Below genes are also the HLA antigen) Class II molecules
− A,B,C (Major gene)
Gene Products: Reactions considered as MHC RESTRICTED:
− HLA A 1. Antigen presentation
− HLA B 2. T and B cell cooperation
− HLA C 3. Cytotoxic T cell interaction with target cells
− E, F, G, H, X (Minor gene) *without MHC these processes will not happen

‒ MHC I and II molecules: essential for cell-mediated Application of HLA typing/Matching:

immunity and therefore appeared at the inception of the 1. Organ transplantation
adaptive immune system 2. Paternity testing
‒ Best organ donor: Siblings or Twinny 3. Forensic medicine, anthropology
4. Studies of racial ancestry and migration
‒ Other products of MHC genes: 5. For diagnostic and genetic counselling
 MHC Class I, II, III 6. Basic research in immunology
 Some Serum
 Some Cytokines Diseases associated to HLA
‒ Organ transplantation ▫ HLA-B23: Ankylosing Spondylitis (Reiter’s
 Tissue typing: Determine/ matching the HLA syndrome)
type ▫ HLA-B47: Congenital adrenal Hyperplasia
▫ HLA-B5 : Behcet’s disease
PROPERTIES OF MHC MOLECULES ▫ HLA-CW6: Psoriasis Vulgaris
▫ HLA-DR3: Type 1 Diabetes mellitus
CLASS I CLASS II
▫ HLA-DR4: Rheumatoid Arthritis
GENETIC LOCI HLA- A, B, & C HLA- SB(DP), ▫ HLA-DR5: Kaposi’s sarcoma
DC(DQ), DR Gold-induced nephropathy
Chronic lymphatic leukemia
CHAIN α chain + β2 α chain + β chain
STRUCTURE microglobulin Methods for detecting HLA antigens: Histocompatibility
testing
*B2 microglobulin is
A. Tissue typing
located in
Chromosome#15
a) Serological approach
i. Lymphocyte Microtoxicity methods
CELL All nucleated cells APC (macrophage, ─ for determination of class I antigens
DISTRIBUTION (found all over the dendritic cells, b) Cellular approach
body except RBC) langerhan cells, T & B i. Mixed lymphocyte reaction-
cells, endothelial and ─ for determination of Class II antigen
stromal cells)

PRESENTS T- Cytotoxic T-helper cells

ANTIGEN (CD 8+) (CD4+)

SOURCE OF Proteins made in the Endocytosed plasma

PEPTIDE cytosol membrane and
FRAGMENTS extracellular proteins

POLYMORPHIC α1 and α2 α1 and β1

DOMAINS
MAJOR HISTOCOMPATIBILITY COMPLEX (MHC)

 Large multi-gene locus consisting of several thousand kilobase pair of DNA on a single chromosome.
 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.
 In humans, it is referred to as Human Leukocyte Antigen (HLA) complex located at chromosome #6

-Chromosome #6 – contains MHC genes ---- major product is HLA

---- also produces MHC CLASS I and CLASS II molecules
---- serum proteins
---- some cytokines and enzymes

NATURE OF HLA ANTIGENS:

1. Glycoprotein component of cell membrane
2. Present in all nucleated cells
3. Products of the genes of MHC

 Inheritance of HLA genes – controlled by MHC located in the short arm of chromosome#6

 Major regions in the MHC: D, B, C, and A

ORGANIZATION BASED ON THEIR: TRAITS UNDER THEIR CONTROLL:

BIOCHEMICAL COMPOSITION:

1. CLASS I – A, B, C, E, F, G, H, X -Transplantation antigens, serologically detected

membrane antigens, Cellular target antigens for cell
Mediated lympholysis.

2. CLASS II – DP, DQ, DR, DM, DN, DO - I Region leukocyte antigen

- T and B cell interaction
- Immune response
- Mixed leukocyte Reaction
- Graft versus Host reaction
- Tumor virus susceptibility
- Peptide transport
- Generation of peptides from cytosolic proteins

3. CLASS III – complement components, - Serum protein molecule

Cytochrome p450, - complement levels
21-hydroxylases and TNF - cytokines and enzymes
(Tumor Necrosis Factor)
 PROPERTIES OF MHC MOLECULES
CLASS I CLASS II
1. Genetic loci HLA-A, B, and C HLA SB (DP), DC, (DQ), DR

2. Chain structure a chain + β2 microglobulin a chain + β chain

(found in C#15)

3. Cell distribution all nucleated cells APC (macrophage, dendritic cells,

Langerhan cells, T and B cells,
endothelial, epithelial and stromal
cells)

4. Presents antigen to T-cytotoxic cells (CD8+) T-helper cells (CD4+)

5. Source of peptide fragments Proteins made in the cytosol Endocytosed plasma membrane
and extracellular proteins

6.Polymorphic domains a 1 and a 2 a 1 and β 1

 The highly polymorphic genes within the MHC gene segment of DNA code for a wide range of cell
surface structures that T-cells must recognize in association with foreign antigen for a successful
immune response.
 MHC Restriction or MHC Restricted Recognition – The process whereby the MHC control
interactions between cells. It involves the recognition of foreign antigen in association with Class I or
Class II molecules.

REACTIONS CONSIDERED AS MHC RESTRICTED:

1. Antigen Presentation
2. T and B cell cooperation
3. Cytotoxic T cell interaction with target cells.

APPLICATION OF HLA TYPING / MATCHING:

1. Organ Transplantation 4. Studies of Racial Ancestry and Migration
2. Paternity Testing 5. For diagnostic and Genetic counselling
3. Forensic Medicine, Anthropology 6. Basic research in immunology

Diseases that are connected in HLA:

HLA DISEASES
 HLA-B27 -------ANKYLOSING SPONDYLITIS for REITER’S SYNDROME
 HLA-B47 -------CONGENITAL ADREANAL HYPERPLASIA
 HLA-B5 ---------BEHCET’S DISEASE
 HLA-CW6 ------PSORIASIS VULGARIS
 HLA-DR3 -------TYPE 1DM
 HLA-DR4 --------RHEUMATOID ARTHRITIS
 HLA-DR5 --------KAPOSI’S SARCOMA
GOLD-INDUCED NEPHROPATHY
 CHRONIC LYMPHATIC LEUKEMIA

METHODS FOR DETECTING HLA ANTIGENS: HISTOCOMPATIBILITY TESTING:

A. 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 (Polymerase Chain Reaction, FRLP (Restriction Fragment Length
Polymorphism)
IMSE WEEK 2 MIDTERM
- Antibodies - Glycoprotein substances synthesized by
plasma cells response to antigenic stimulation.
 IgE is the most uncommon or rare because it is
only seen with allergic reaction and defense
against parasites

 not a pure protein but rather a glycoprotein

 Made by plasma cells (activated B-cells), B-cell A. According to its sedimentation constant
will only be activated with a corresponding
antigen IMMUNOGLOBUL SEDIMENTATI MOLECUL
 also known as immunoglobulins (made up of INS ON AR
gamma globulin fraction of the serum proteins) COEFFICIENT WEIGHT
(DALTON)
o Immunoglobulins is the heaviest that’s
IgG 7s 150,000
why in electrophoresis it is considered
(lightest)
the lowest fraction of gamma globulins.
Serum IgA 7s 160,000
o Albumin, on the other hand is
Secretory IgA 9s; 11s; 13s 170,000
considered as the fastest serum protein in IgM 19s 900,000
electrophoresis (heaviest)
 2-8% carbohydrates and 82-96% protein IgD 7s 180,000
 All antibody are immunoglobulins but not all IgE 8s 190,000
immunoglobulins are antibody
o Immunoglobulins and antibodies are
substances that can bind with antigen; Sedimentation constant/coefficient
however if the antigenic specificity is not  Unit used to express the rate of sedimentation of
yet known it is considered as immunoglobulins when they are subjected to
immunoglobulins but if it knows where analytical ultracentrifugation
specific antigen to bind then it is antibody o When you centrifuge a serum or plasma
samples containing immunoglobulins or
o Only when the immunoglobulins have the antibodies their settlement in the
specific antigen to react with that it can sediment have a rate. The unit use is ‘s’
be called as antibody known as Svedberg unit
-Structurally all antibodies are immunoglobulins but Classify immunoglobulins as:
functionally not all immunoglobulins are antibody.
Monomer – basic unit of antibody (ex. IgG, IgD and IgE)
-Immunoglobulins are considered to be the humoral
branch of the immune response o Polymer - When an antibody consists of
more than 1 monomer
PROPERTIES OF AN ANTIBODY:
Dimer – 2 monomers (ex. Secretory IgA)
1. Protein in nature (Glycoprotein)
2. With high molecular weight Trimer – 3 monomers
3. Present in serum/plasma, saliva, semen, CSF and Pentamer 5 monomers (ex. IgM)
other body fluids.
o Dominant with serum or plasma  Serum IgA – can be a monomer, dimer and trimer

CLASIFICATION OF ANTIBODIES: IgG, IgA, IgM, IgD , IgE Classify antibodies according to the temperature at
(IgG most abundant; IgE least abundant) which they react

 Arrange in decreasing concentration In the 1. Cold antibodies – react best at 4C to room

serum temperature (24-27C)
 o Ex. ABO antibodies (doesn’t need to
incubate to an incubator)
2. Warm antibodies – 37C
o Rh antibodies (need to incubate to
enhance the reactivity of Rh antibodies at
37C)
Classify according to occurrence:
1. Natural antibodies – antibodies produced even
without previous antigenic stimulation
- ABO blood group system - Type A patient
automatically have anti-B antibody,
because it is naturally occurring and
doesn’t need to be exposed with type B
blood to acquire anti-B antibodies
o Ex. ABO antibodies
2. Immune antibodies – also known as the acquired
antibodies. Antibodies that can only be produced
given that there is exposure with corresponding
antigen
- Rh blood group system – there is two blood
type:
 RH Positive – the red cells have D antigen
-Rh positive have D antigen that’s why
it cannot have Anti-D antigen or else it
will lead to destruction of cell
 Rh negative – no D antigen in the red cell
-Only Rh negative can produce anti-D
antibody
-Doesn’t produce anti-D antibody not
until it is exposed to a Rh positive or when a
mother is pregnant with a Rh-positive baby
o Ex. Rh antibodies
Classify according to the reaction with an antigen
 In suffix for antibody
 nogen suffix for antigen
 Agglutinins – involved in agglutination reaction
o Agglutination (antigen involve is cellular
or particulate) Big or attached to the cell
o An antigen that is involved in the
agglutination reaction is called
agglutinogen
o Blood typing (Ex. of agglutination
reaction)
o Anti-sera
 Precipitins – involved in precipitation reaction
o Antigen involved in precipitation reaction
is called precipitinogen
o Precipitation (antigen involved is soluble
antigen)
 Agglutinoids – agglutinins that are modified by
HEAT (agglutinins become agglutinoids when
heated)
 Hemagglutinins – antibodies that can clump RBC
 Lysin – antibodies that can cause lysis
o Hemolysin – lysed RBC
o Leukocydin -lysed WBC
o Bacteriolysin – lysed bacteria
 Opsonins -can coat microbes for easier
phagocytosis (Ex. antibody or immunoglobulins)
 Neutralizing antibodies – antibodies that
neutralizes certain reaction
 allergic antibodies – involved during
hypersensitivity reaction
 antitoxins – neutralized harmful effect of toxins
 complement fixing bodies – has the capacity to
activate compliment system
According to their In Vitro behavior

1.Complete 2.Incomplete
Classify according to the species that produces them antibody (IgM) antibody (IgG)
o First to o produce
1. Isoantibodies/alloantibodies – antibodies appear by
produced in response to antigen of the other after memory B
individual of the same species infection cell
o Ex. Blood transfusion and organ Synonyms Bivalent; saline Univalent;
transplant acting blocking;
2. Heterophile antibody – antibodies produced coagglutinating;
conglutinating
in response to antigen from other species
Response to Thermolabile Thermostable
temperature Disulfide bonds – chemical bonds essential for the
Ability to cross Cannot cross the Can cross the normal three-dimensional structure of immunoglobulins.
placenta placenta placenta  The type of bonds that connects the different
Occurrence Early in Late in parts of the antibody monomer
immunization immunization Two types of disulfide bonds:
reaction Saline acting Albumin acting  Interchain – connects light chain and heavy
chains
The structure of immunoglobulins  Made up of sulfur molecules
Intrachain – connects domain of the heavy chain and also
Monomer – basic structural unit of antibody and are connects the domains of the light chain
made of tetrapeptide chain (4)

Tetrapeptide made up of:

1. Two Heavy chain – with five principal antigenic types
with their corresponding immunoglobulins (gamma,
alpha, mu, delta, epsilon)
 The class of the immunoglobulins depends
on the heavy chain it contains
 Gamma -IgG; Alpha – IgA; mu – IgM; Delta –
IgD; epsilon - IgE
 The two heavy chain must be identical
(ex.gamma – gamma)
2. Two Light chain – with two antigenically defined types:
kappa and lambda
Domains – globular regions on polypeptide chain
stabilized by intrachain disulfide bonds.
Domains on the heavy chains – VH (variable
region of heavy chain), CH1, CH2, CH3, (CH4)
 IgE have an extra domain (CH4)
that’s why it has a higher molecular
weight and sedimentation conc.
Than other monomer
Domains on the light chain – VL
(variable region), CL (constant
region)

Hinge region – flexible part of the antibody

located in the heavy chains because it contains
the enzyme proline, as a result the antibody
molecule can bend (monomer). It is more
exposed to enzyme and chemicals thus papain
acts here to produce Fab and Fc fragments. The
light chains are each link to one-half of a heavy
chain by disulfide bonds at the proximal end.

The middle part that was bended, and

rich with amino acid proline.

Paratope –
binding site of
antibody for
antigen
VL + CL  binds with antigen  IgM, a pentamer. Because it has five
monomers then it contains 10 binding
CH1  binds with C4b
sites for antigen.
CH2 binds with C1q (if IgG)  IgM is the most powerful
antibody in causing
CH3  binds with C1q (if IgM) also the binding site for T agglutination and precipitation
and B cells, platelets and mast cells, monocyte and reaction.
macrophage  Polymer -immunoglobulin composed of more
CH2 + CH3  binds with NK cells, placental than a single basic monomeric unit
syncitiotrophoblast and neutrophil  J chain – Polypeptide chain which normally holds
polymeric immunoglobulins
 It comprises the Fc part of antibody  Secretory components – a substance attached to
polymeric immunoglobulins found on secretions
(present only in secretory IgA)
Function of antibodies: o Protects secretory IgA from acid digestion
 Binding with antigen – to destroy antigen in the stomach
 Opsonization – phagocytes such as neutrophil, o Facilitated the transport of secretory IgA
monocyte and macrophages binds with the Fc across mucosal surfaces
portion of antibody  Secretory IgA is the major
 Complement fixation and activation – there are antibody for mucosal
some domain where complement binds defense/surface
 Respiratory tract
 Gastrointestinal tract
 Genitourinary tract
Regions on polypeptide Chains:

Variable region Constant

A. Amino acid o Amino acid is Fragments of immunoglobulins:
sequence subject to fixed and
changes unchanging  2 Fab (antigen binding fragments) -consist of one
 Binding site light chain and ½ heavy chain
for the  Binds with antigen
antigen, the  1 Fc (Crystallizable fragment) -made up of two
paratope of half heavy chain
antibody  Binds with host cell such as
binds with phagocytes and compliment
epitope of
components
antigen
B. Amino terminals o Carboxyl Enzyme Digestion
(NH2) positive + terminal (COOH)
negative -  Studying antibody structures, you can use various
C. Concerned with o Concerned with enzyme like:
binding antigen binding to host  Reason why you need to use these
various enzymes is so that you can study
part by part the components of antibody
o The amino terminal end of antibody has a
o Cleavage with papain enzyme – cleaves
positive charge while the binding site of antigen
before the hinge
(epitope) have a negative charge that’s why it
 Product after papain digestion is
forms ionic bond causing an affinity giving
two Fab + one Fc
specificity.
o Cleavage with pepsin enzyme - cleaves after the
o An antibody is a monomer having two binding
hinge
sites for antigen.
  Pepsin digestion will to lead to one
F(ab)2 – A two Fab that is combined
together by the hinge and used to
indicate thar it doesn’t contain the hiinge
o Reduction using mercaptoethylamine
/mercaptoethanolamine
 It disrupts disulfide bonds and when
destroyed it produce two heavy chain and
two light chain
Genetics of immunoglobulins:
 Genes coding for the synthesis of heavy chains
are located in chromosome no.14 (gamma, alpha,
mu, delta, epsilon)
 Genes coding for the synthesis of kappa light
chains are located in chromosome no.2
 Genes coding for the synthesis of lambda light
chains are located in chromosome no.22
‘THE GENES INVOLVE WITH THESE CHROMOSOME ARE
INVOLVED WITH THE SYNTHESIS OF PROTEINS’
With these chromosomes it can be found in:
 V gene - variable
 D gene - diversity
 J gene - joining
 C gene - constant
Immunoglobulin Variability:
1. Isotopic variation (isotypes) -refers to the
different heavy and light chains classes
o Different antibody classes differ on the
heavy chain and light chain content
o Ex. IgGAMED
2. Allotypic variation (allotypes) – refers to the
genetic variation within a species involving
different alleles at a given locus
o Subclasses within a class of
immunoglobulins
3. Idiotypic variation – refers to the diversity at
the binding site and in particular relates to
the hypervariable segments of the antibody
combining site (paratope)
o Antibodies to different antigens
Characteristics of the different type of immunoglobulins:
1. IgG (when the patient is healing IgG increase;
marker of puss or chronic infection)
- Predominant immunoglobulins among
humans comprising 75-80% of the total
immunoglobulin pool. (most abundant)
- Major antibody produces by the memory
cell
- Has four major subclasses: IgG1, IgG2,
IgG3, IgG4 (arrange in decreasing
concentration in the serum) being IgG1 as
the most abundant
o the reason it has different
subclasses is that it has a minor
variation within the gamma chain
- Equally distributed in the different fluid
compartments with detectable amount in
CSF and urine
- Readily diffusible – highest diffusion
coefficient, only one that cause placenta
and can cause HDN in the fetus
- IgG antibody response appears later than
IgM in primary response but they form
the major antibody of the secondary
immune response
- Maternal IgG is actively and selectively
transferred across the placenta to the
fetus and imparts passive protection to
the newborn for 6-9 months
- Major antibody of the secondary
immune response/anamnestic immune
response
o Occurs when the same antigen
are present; increase more if it’s
the same antigen
- Marker of lifelong immunity; protects
from the same antigen
- Functions of IgG:
 Provides immunity for the
newborn
 Complement fixation – IgG3, IgG1,
IgG2 (IgG1 as the most efficient
complement)
And for IgG4 cannot activate the
complement via classical
pathway
 Opsonization
 Neutralization of toxins and
viruses
  Participation in agglutination and
precipitation reactions
- Need booster to prime again the memory
B cell (1 shot)
2. IgM (the first to response but will decrease level
when the patient is already healing)
- The largest of the immunoglobulin
molecule, accounting for 5-10% of the
total immunoglobulin pool.
- Star-shaped in the free state; crab like in
antigen-antibody reaction
- The earliest antibody to appear in the
primary immune response but it does
not persist for long (major antibody for
primary immune response)
- Maternal lgM does not cross the placenta
- IgM detection in newborn is a useful
indicator of intrauterine infection
- A powerful agglutinator of a particulate
antigen
- A marker for active, acute or present,
current infection
- Functions of IgM:
Complement fixation
Agglutination
Opsonization
Neutralization of toxins
Surface receptor for antigens
(on B-cells)
3. IgA
- Represent 15-20% of human serum
immunoglobulin pool
- Found in serum in small amount but
predominant in sero-mucous secretion of
the respiratory tract and gastrointestinal
tract. It is also found in tears, sweat,
saliva, colostrum and breastmilk.
- Forms IgA: (allotypes)
Serum IgA (IgA1) – can
agglutinate motile infectious
agents thus promoting their
phagocytosis but they cannot
THEORIES OF ANTIBODY REACTION
activate the complement system
Secretory IgA (IgA2)– a polymeric
form stabilized a short
polypeptide chain. It is known as
the ‘antiseptic paint’ of mucous
membrane. It can activate the
bacteriolytic activity through the
alternate pathway of
complement system nd only in
the presence of lysozyme
4. IgD
- heat labile immunoglobulin, accounts for
less than 1% of the total serum
immunoglobulin but is known to be
present in large quantities on the
membrane of many circulating
immunocompetent B lymphocytes.
- Detection by highly sensitive assay
requiring radio-labelled antisera
- Precise biological action is not known but
it may play a role in antigen-triggered
lymphocyte differentiation
- No known function except it is a marker
for mature B-cells (B-cell differentiation)
5. IgE
- Heat labile immunoglobulin. Least
abundant Immunoglobulin in the serum
accounting for only 0.004% of the total
serum immunoglobulin
- Synthesize locally by plasma cell present
in the mucous membrane of the
gastrointestinal tract and respiratory
tract
- It is unable to fix the complement via the
classical pathway
- It is homocytotropic due to its affinity for
cells of the hosts species, particularly for
tissue mast cells and blood basophil.
- Because of its ability to attach to the
human skin, it is associated with
immediate hypersensitivity reactions
but also, apparently, with immunity to
certain helminthic parasites
- Also known as reaginic
antibody/nuisance antibody
1. Paul Ehrlich’s side chain theory
 Certain cells have specific surface
receptors for antigen that were present
before contact with antigen occurred.
 Once antigen was introduced, it would antigen would continually increase
select the cell with proper receptors, lymphocyte pool
combination would take place and then  Most acceptable among all theories
receptors will break off and enter thr  When an antigen goes inside the body it
circulation as antibody molecules. New will select an appropriate clone of B cell
receptors will be formed in place of those it will cause to activate the B cell
broken off and this process could be producing plasma cell and memory cell
repeated. and produces antibodies
 On the surface of the cell there are
receptors for the antigen, and when an
antigen are inside the body and challenge
the immune system it will cause the
receptor to break apart or falling out the
cell and becoming antibodies

2. The temple theory/instructive theory (Felix

haurowitz)
 Antibody-producing cells are capable of
synthesizing a generalized type of
antibody and when contact with an
antigen occurs, the antigen serve as a
mold or template and alters protein
synthesis so that antibody with a specific
fit is made. The ‘molded’ antibody that
enters the circulation, while the antigen
remains behind to direct further
synthesis
 All antibodies inside the body are in
standard form: unfolded gamma globin
 All antibodies inside the body are the
same the antigen is the one that will
serve as the template to form the
antibody, it means the antigen will direct
the formation of antibody
3. Selective theory
(replicationtranscriptiontranslation)
 Assumes that antibodies are synthesized
in a manner similar to that other proteins.
Instruction for their synthesis are provide
by genetic elements in the nucleus of the
cell rather from the antigen
4. Clone selection theory (niel jerne and mcfarlane
Burnet)
 Individual lymphocytes are genetically
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
proliferate. Repeated contact with the
COMPLIMENT

 Collective term designating a complex series/mixture of plasma proteins that have functions of zymogen.
o Group of non-Ig circulating in the blood in biologically inactive form (zymogen)
 Inactive form (zymogen)  active form (serine protease) having a enzymatic abilities
because most of the compliment are capable of acting as serine protease.
o Compliments the action of antibody in destroying the antigen
o Sometimes, antibody itself cannot destroy antigen on its own and It might need help with other
components, just like compliment.
o Made up of more than 30 globulins/protein that are normally present in the body and it could b
either alpha or beta globulins
 *As the antibody are made up of gamma globulins, compliment however is made up of
either alpha or beta globulins. *
o The compliment system is made up of: Major components
 Arrange according to its discovery:
 C1
 C2
 C3
 C4
 C5
 C6
 C7
 C8
 C9
 Regulatory and inhibitory proteins of the compliment system – The rest of the protein
that comprises the compliment system
 Viral function:
1. Host defense mechanism
- Opsonization
- Chemotaxis and leukocyte activation
- Lysis of bacterial and mammalian cells (end product)
- Stimulation of inflammatory response
2. Interface between innate and adaptive immunity
- Augmentation of antibody response
- Enhancement of immunologic memory
3. Disposal waste
- Clearance or removal of immune complexes from tissue (spleen)
 Before the spleen takes up or destroy the immune complex there must be the
presence of compliment
 When there’s deficiency of compliment in the body there’s a possibility of
immune complex accumulation and might form deposit in the body triggering
inflammatory reaction hence causing tissue damage.

 Immune complex = antigen – antibody complex

 Complimentary alphabet is formed when compliment system is active those forming fragments
ROUTES OR PATHWAY OF COMPLIMENT ACTIVATION
Classical – proteins term as components are symbolized by the letter C followed by a number
  3 stages:
 Initiation or recognition
 Amplification or enzymatic activation
 Membrane attack leading to cell destruction
 Immunologic activators: immune system

li

When immune complex is been formed it sends signal to the to C1 then C1 subunit will be cleave
(separated). The C1q is also known as recognition unit of the classical pathway, because it was
the first compliment fragment will be formed and will initiate the chain reactions of compliments
proteins.
When C1q is activated it will activate C1r. when C1r is activated it will be called C1r (C1r Bar) or
(activated C1r), then it will activate C1s then activated C1s will be written as C1s and read as
(activated C1s).
Activated C1s. has a capacity to cleave C4 and C2. In the compliment fragment the one having
the (a) designation is the one having a smaller subunit and the one having the (b) designation is
the one having a larger subunit.
The C4a has no role in the activation process and served as an Anaphylatoxin.
Anaphylatoxin: C4a, C3a, C5a: Mediators of Inflammation
C4a Mediates Inflammatory Reactions.
The C4b and C2a will be combined then form C4b2a and the one that stabilize the combination is
the magnesium ions and known as C3 Convertase of the Classical pathways of the Compliment
System.
C3 convertase will act upon on the C3 compliment molecules. And C3 will cleave C3a and C3b.
C3a will have no role in the activation then it will go to the plasma and also act as Anaphylatoxin,
and C3b act as an Opsonin then C3b will join C4b2a and formed C42a3b and called as C5
convertase of the classical pathways.
As the term implies an C4b2a3b as an enzyme then it will cleave the C5 then C5 will cleave C5a
also has no role in activation it will also join Anaphylatoxin and be a chemotaxin. Then C5b will be
bind into a cell, the cell has an antibody (IgM, IgG) then start creating a force on the cell. Then
the immune compliment fagus will be produce. If the C5b is finally produce the C5b will deposit
on the surface of bacterial cell, then C5b will make a hole in the bacterial cell to make the C5b
bond stronger the C6 and C7 will be combine to C5b then formed C5b67 after C5b67 is formed
it will be combined with C8 and C9 and form C5b6789 and became a MAC (Membrane Attack
System), and when the C5b6789 is formed in the surface of the bacterial cell or any cell it will be
the MAC then causes Lysis of the cell or (Cytolysis), then cell will be Lysed, then your
Compliment System help your antibody to destroy the cell.

Alternative/Alternate/Primitive/Bypass/Properdin – normal serum proteins are termed as factors and

are symbolized by letters essential in the initiation of this:
 Soluble C3
 Factor B - analogous to C2
 Factor D – believed to be similar with C1s
 Properdin – a gamma globulin which when complexed with C3b alternate pathway C3
convertase
It was called Bypass because the activation process starts at C3 and Bypass C1, C4, C2.
The major activators are non-immunologic in the second pathway. The immune complex could be also an
activator (IgA,IgA4 and IgF) but the major activator of the alternative path way are the non-immunologic
such as LPS (Lipopolysaccharide), and LPS is always present in all gram negative bacteria and some
viruses like HIV can directly activate the alternative pathway and some parasites like trypanosomes,
Schsitomes and under this pathway there is no C1,C2,C4 because it begin in C3.
In the Alternative pathway there is a phenomenon called Tick Over, Tick Over is a spontaneous activation
of C3.
C3 is the central component of the compliment system because in the C3 is always had a same pathway
whether it was classical, alternative or MBL pathways are the same when they reach the C3 pathways.
Mannose binding lectin (MBL) pathway – a member of the so called collectin family of molecules;
structurally similar to C1q and it functions as opsonin. In this pathway, the interaction of MBL with
carbohydrate on the surface of polysaccharides of microbes leads to the formation of enzymatic
complex that binds and activates C4 and C2
CLASSICAL ALTERNATIVE
Immunologic activators: Immunologic activators:
IgM, IgG3, IgG1, IgG2 bound to antigen = immune Aggregated IgA and in some instances IgG4 and IgE
system
Non-immunologic activators: Non-Immunologic activators:
Apoptotic cells, Staphylococcal protein A, CRP bound Bacterial and plant polysaccharide, LPS, zymosan,
to ligand, certain viruses and gram-negative bacteria, inulin, cobra venom factor, viruses and tumor cells,
DNA some parasites like trypanosomes
Possess a recognition unit No recognition unit
Requires presence of calcium and requires C1, C2, C4 Lack of dependence on calcium and activation
for its activation immediately starts with C3 (bypass pathway)

MAIN SOURCE OF COMPLEMENT PROTEINS: Hepatocytes, intestinal (C1) and urogenital epithelial cells, blood
monocytes ang macrophages.

 Many compliment molecules genes are localized in the MHC (C4 and C2) MHC class 3 genes
 Homeostatic maintenance of complement activation is mediated by regulatory proteins.
o Ex.
- On plasma: anaphylatoxin inhibitor, C1 inhibitor, Factors H and I, C4 binding protein, S
protein and S 40-40.
- On cells:C3b/C4b receptor (CR1), decay accelerating factor, membrane co-factor protein
and CD 59.
o Membrane inhibitor of reactive lysis (CD59)
 In-vitro destruction of complement: how to produce inactive serum
o Addition of chelating agents (EDTA)
- (EDTA) Ethylenediamine tetra acetic acid
- The EDTA will cause a decomposition of C1 and C1q, C1r, C1s, will not be formed
and it will not form an activation or the activation process will be slowed.
o Heat Inactivation of serum – heat serum at 56 degree Celsius for 30 mins.
- Heat the serum at 56 degree Celsius for 30 minutes.
- If you did that you are destroying all the compliments in the serum because
proteins are denatured at high temperature.
o Treatment with zymosan
- Add Zymosan as a chemical inactivator of Compliment.

 Nephelometry- an instrument used to measure the compliment proteins in the blood.

 Inactivated serum – serum sample where complement proteins are destroyed

 Diagnostic evaluation of complement:

o Serum/plasma assay of C3 and C4 by nephelometry
o C1 binding assay measures the binding immune complexes containing IgG1, IgG2 or IgG3
and/or IgM to C1q. it can be useful as prognostic tool to diagnosis at during remission of
acute myelogenous leukemia

AH50
CH50
Deficiencies of Compliments Components
Having a deficiency in C1, C2, and C4 could lead to Lupus like Syndrome.
Lupus is an autoimmune disease where in a body a person with lupus has also auto antibody/antibodies.
When we say auto antibodies it reacts to your own cells or become antigen.
If a person had a Lupus Syndrome the person had a many immune complex that reacts into auto
antibody.
If somebody had a deficiency in C1, C2, C4 the spleen can’t take up the formed immune complex and
that immune complex will accumulate in your blood. Like with someone has a lupus syndrome.

COMPLEMENT FIXATION TEST -secondary immunologic/serologic test

 2 system:
o Test system – contains antigens and antibodies. One of each is unknown so if antigen is
unknown you will use antibody for that specific agent.
o Indicator system – sheep RBC coated with hemolysin
 Serologic Test- it was an antigen and antibody reaction.
o Ex. Measuring HIV Antigen you will use Anti-HIV Antibody as a Reagent.
If the antigen is not being present use the specific antibody to find the unknown antigen. If the unknown
is present you will add External Source of Immuno Compliment and It will combined with the postive or
present antigen or antibody.
If the antigen or antibody is not present or negative and you add external source of immuno compliment
it will lyse the SRBC because indicators need an External source of Immuno compliment or Immuno
compliment to perfom or form a Lysis/lysed cell.

External source of Compliment-guinea pig serum is a good source of compliment.

Indicators- Sheep RBC Coated hemolysin.
IMMUNOMODULATION

 Modifying the immune response

 Deliberately or willing fully modify the immune response. To enhance or suppressed.
 Positive immunomodulation:
o Immunoenhancement/immunopotentiation
 Negative immunomodulation
o Immunosuppression
 Disabling the immune system for some reason
 Disable the immune system whenever there’s hypersensitivity or allergic
reaction to a food, so that later on you can fucking enjoy eating whatever you
want
 Suppressed the immune system of the recipient with immunosuppressant drug
so that in transplantation, temporarily, it can assure that there would be no
rejection of an organ. Because even if the recipient and donor had the same
tissue and HLA type there would be still a slight risk for organ rejection.
- Immunosuppressant drug is given after the transplantation

Immunoenhancement

 Accomplished through the addition of adjuvant to our vaccines

o Adjuvant are added to the vaccine to increase the size of antigen in the vaccine, the
bigger the substance the more its immunogenic. If the antigen is bigger then it would
result to prolong exposure of antigen in the immune system making the lymphocytes,
antibody producing cell, plasma cells will be having an easier time to respond to that
antigen, making them be more efficient in producing antibodies in a much faster way.
 Components of adjuvant:
o Muramyl dipeptide – major component of adjuvant that is available in the market
o Cell wall constituent of killed/attenuated microorganisms (M. tuberculosis, B. pertussis,
Brucella spp.)
o Other components: mineral oil, lanolin, alum, detergent, acrylic particles,
polynucleotides
 Example of adjuvant:
o Aluminum hydroxide
o Complete Freud’s adjuvant (made up of killed M. tuberculosis, mineral oil, lanolin)
o Incomplete Freud’s adjuvant – same component with complete Freud’s except that it
does not contain a killed M. tuberculosis
 Containing only of mineral oil and lanolin
Immunosuppression

 Negative type of immunomodulation

 Suppressing the immune system/immune response so that it won’t react anymore to a certain
antigen
 Two types of immunosuppression:
o Specific
o Non-specific
 Artificially induced
 Physical means
 Chemical means
 Biological means
 Naturally induced
 Immunodeficiency disorder that could be inherited or acquired
 Specific immunosuppression
o Refers to immune tolerance
o Tolerance:
 Inability of the body to mount immune response to a substance that is
potentially immunogenic
 When the body develop immune tolerance, it means it cannot respond to
antigens and as a result you are prone to a variety of infection
o Secondary nothing response
 Normal: During a repeat infection the immune system mounts a faster response
because during the first infection the body produces memory cells already.
 Ex. measles virus
 When it comes to secondary nothing response instead that it can act faster
towards an antigen that’s coming back causing repeat infection, the immune
system won’t respond to it.
 Nonspecific immunosuppression (artificially induced)
o Physical means
 Irradiation – can destroy cells in the body. Chemotherapy, its side effect is not
only it destroy cancer cells but also healthy cells. That’s why patient becomes
immunodeficient, becomes anemic and suffer from bleeding
 Thoracic duct drainage
 Thymectomy – removal of thymus. It’s very important because it is the primary
lymphoid organ for T-cell. Where T-cells mature.
 Splenectomy – a secondary lymphoid organ, the activation site for the
lymphocytes. Where b and T lymphocytes are activated. B-cells cannot
transform into memory cell or plasma cell. Whereas, antibody will be produced.
 Bursectomy
o Chemical means
 Corticosteroids – steroids are immunosuppression drugs
 Alkylating agents (cyclophosphamide)
 Anti-metabolites
  Antibiotics (cyclosporine A) – cyclosporine A can cause aplastic anemia and also
immunosuppression
o Biological means
 Anti-lymphocyte antibodies
 Anti-thymocyte antibodies
 Antigen desensitization -allergic reaction. Expose yourself to allergen gradually
(small doses) until you develop resistance to a allergen that causes allergy to
you.
 Nonspecific immunosuppression (naturally induced)
o Examples are different immunodeficiency disorders
 Congenital
 Hereditary
 Acquired
 Immunodeficiencies
o Deficiency of the immune system include:
 Phagocytic cell deficiency
 B lymphocyte deficiency
 T lymphocyte deficiency
 Combination of T and B lymphocyte deficiency

PHAGOCYTIC CELL DEFICIENCY

-result in decreased ability to phagocytized and kill bacteria

a) Chronic granulomatous disease

- A genetic disease characterized by ineffective killing of bacteria by
neutrophils.
- Patient with CGD, their neutrophil is unavailable to kill bacteria specially
those that are catalyze positive.
- CGD is caused by the defect in cytochrome B oxidase, which results in
hydrogen peroxide production. Hydrogen peroxide is necessary for
producing the toxic superoxide that are critical in bacterial killing.
Hydrogen peroxide are used by the phagocytes to kill catalase-positive
organism.
- Diagnosis. The nitro blue tetrazolium (NBT) reductase test is used to
detect impaired neutrophil phagocytosis. The neutrophils of CGD patients
fail to reduce the NBT dye.
- Symptoms. Patients with CGD suffers from recurrent infections caused by
catalase-positive bacteria and yeast and fungi.
- Treatment. Includes the use of GSM-CSF or G-CSF and IRN-y.
b) MPO deficiency
- Inherited as an autosomal recessive trait and is one of the most common
inherited disorders. absence of an enzyme myeloperoxidase (digestive
enzyme) will not kill pathogens. The MPO in the primary granules of
neutrophils is decreased or absent, and although phagocytosis takes place
normally bacterial killing is inefficient. Fungal killing is more seriously
impaired than bacterial killing, although otherwise healthy patients with
MPO deficiency do not have an increase frequency of infection, diabetic
patients who have this disorder may have an increase in Candida spp.
infections.
c) Cr3 (ic3b receptor) deficiency
- A rare autosomal recessive trait characterized by a decreased or absence
of specific complement component receptors on neutrophils, monocytes
and lymphocytes. These receptors are responsible for adherence-related
function. Abnormalities result in defective margination and diapedesis of
neutrophils, impaired chemotaxis, and ineffective phagocytosis. T
lymphocytes adhere poorly to target cells. Clinically, there is an increase
frequency of bacterial infections, a decreased inflammatory response and
neutrophilia.
d) Specific granule deficiency
- Inherited as an autosomal recessive trait. Neutrophils fails to develop
specific granules during myelopoiesis, and as a result patient who have
this disorder experience severe recurrent bacterial infections.
e) Chediak higashi syndrome
- An inherited disorder that is characterized by abnormal fusion of primary
granules in neutrophil. During phagocytosis, degranulation is impaired
and little or no MPO is released into the phagosome. Patients who have
chediak syndrome have recurrent bacterial infections and are also
characterized by albinism and extreme photosensitivity.
- You have a granule content on the neutrophil but the problem is its slow
to adhere in the pathogen that was engulf.
f) Lazy leukocyte syndrome includes:
1. Job syndrome also known as hyperimmunoglobulin E, characterized
by poor chemotaxis and recurrent skin infections and abscesses
2. Tuftsin deficiency. Tuftsin is a chemotaxis that also improve
phagocyte motility, engulfment and oxidative metabolism. Affected
persons experience bacterial infections.
3. Actin dysfunction a deficiency of the cytoskeletal protein actin can
result in decreased bacterial motility and chemotaxis. Patients
experience recurrent bacterial infections.
g) Bruton’s agammaglobulinemia
- A sex-linked disorder that primarily affects men. It is usually recognized
early in life when antibodies fail to develop. Pre-B cells maybe found in
the bon marrow, but they do not mature. Few mature B cells are found in
 the peripheral blood. Gamma globulin levels are markedly decreased. This
disorder may be treated with gamma globin preparations.
- Due to the deficiency of the Bruton’s thymidine kinase, which is very
important in the maturation of B lymphocytes. Would only result to pre-B
cell, resulting to no antibodies.

-
Maturation: pro B cell  pre-B cell  immature B cell  mature b cell 
plasma cell
h) Common variable hypogammaglobulinemia
- Acquired disorder in which 1 or 2 immunoglobin classes are deficient.
Total immunoglobins are normal, because decrease of one immunoglobin
is often compensated by an increase in the production of another.
Selective IgA deficiency is one of the most common of these deficiencies.
typically, only those patients whose disease includes IgG deficiency suffer
from increased bacterial infections.
- Because IgG is the most abundant 75-80% (GAMDE – Decreasing
concentration)
i) Neonatal hypogammaglobulinemia
- Caused by the normal immaturity of the neonate’s immune system. It
corrects itself between the age of 6 and 12 months as infant’s immune
system mature.
- We never perform reverse typing on the serum of neonates because they
don’t have antibodies yet. If ever, it comes from the mother. Antibodies
will be present at 6 to 12 months.

T-LYMPHOCYTE IMMUNODEFICIENCIES
 Without accompanying loss of B-cell function are rare, composing only &5 of
immunodeficiencies. These disorders may be acquired or inherited.

a. DiGeorge Syndrome
- Results when the thymus gland develops abnormally during
embryogenesis. Abnormalities of other endoderm-derived tissues are also
seen. T lymphocytes are usually decreased, but may be normal. Most
patients have high CD4-CD8 ratio. Although antibody responses may be
normal, cell mediate immune responses are impaired.
b. Nezelof syndrome
- An autosomal recessive disorder. Patients are athymic and are especially
susceptible to viral and fungal infections, which can be fatal in these
patients
- Totally no thymus gland (athymic)
COMBINED B AND T LYMPHOCYTE IMMUNNODEFECIENCIES
-The most serious of the immunodeficiencies, because both cell mediated and humoral immune
response are affected.

a. Bare lymphocyte syndrome

- characterized by defects of in class 1 MHC antigen expression, Class II MHC antigen expression or a
combination of both. CD4 positive T lymphocyte are decreased in number, and B and T cell activation is
reduced.
- if you don’t have MHC class I or II, t lymphocyte is affected because it requires antigen presentation in
association with MHC
b. severe combined immunodeficiency disease
- may be inherited as autosomal recessive or x-linked traits. All are characterized by markedly decreased
numbers of both T and B lymphocytes.
c. acquired immunodeficiency syndrome (AIDS)
-caused by the human immunodeficiency virus I (HIV-I) or the human immunodeficiency virus II (HIV-II).
The T-helper cell/CD4 positive T lymphocytes are the primary target cell, which are the central cell of the
immune system. It is the central cell because its major role is to produces cytokines that controls or
regulate the development of all the component of the immune system. Approximately 5% of B
lymphocyte are also affected.
-T helper, helps the phagocytes and help the B cell in producing antibody. So, if its deficient not only the
T lymphocyte is affected but also the functioning of B lymphocyte, function of phagocyte will also be
affected.
- that’s why people with HIV have an overall immunodeficiency.
o human immunodeficiency virus I (HIV-I) – a strain which are known worldwide
o human immunodeficiency virus II (HIV-II)- strain that is common in Africa.
AUTOIMMUNITY

 Conditions in which damage organs and or tissues results from the presence of autoantibody or
autoreactive T cells.

Etiology: (main possible cause) defect in mechanisms underlying self-recognition

 Normally our immune system can distinguish between the foreign and self-antigen but for those
patients developing autoimmunity the immune system losses it’s ability to recognize which is
foreign and self that results to attacking the self-antigen and that organ contains antigen are
being destroyed
 Some autoimmune disease maybe secondary to some microbial infection

Possible mechanism:

1. Loss of self-tolerance by B and/or T cells

2. The response to self-reactive antigens
3. The formation of neoantigen
o a phenomenon molecular mimicry – certain microbes possess antigens that
closely resembles human antigen.
 Ex. Coxsackie virus – contains antibody against glutamic acid hydrolase
(enzyme inside the body)  type 1 DM
- This virus contains an antigen that are similar with glutamic
acid hydrolase
 Ex. Streptococcus pyogenes  glomerulonephritis and rheumatic heart
disease
- The M protein closely resembles the self-antigen in our
kidney and heart
4. Release of maturation antigen (sequestered antigen)
o Can be seen inside the organs that are immunologically privileged
5. Deficiency of T suppressor cells
o Involve in the suppression of autoreactive T cells and autoantibodies
o Insufficient T cell can develop to autoimmune disease

 Most autoimmune diseases exhibit marked familial incidence suggesting genetic predisposition.
 Autoimmune responses are often related to the presence of specific Major Histocompatibility
Complex (MHC)
SOME BODY PARTS ARE IMMUNOLOGICALLY PRIVILEDGED

-Means antibodies and other component of the immune system do not pass inside these organs.

 Brain
 Eye
 Testis – shouldn’t make a severe damage or else the barrier will be damage and resulting to
enter lymphocytes and antibodies inside the testis. And inside the testis is sperm which is a
good example of maturation antigen.
o If a testis were in trauma and a B-lymphocyte goes inside the testis it would
assume that the sperm is a foreign organism and therefore the B-lymphocyte
would produce antibodies against the sperm and because of that the person
could become sterile.
 Uterus (fetus) – there’s an antigen of fetus that are foreign to the mother because not all genes
of the baby are similar with the mother, only half
 Hamster cheek pouch

TWO TYPES OF AUTOIMMUNE:

1. ORGAN SPECIFIC – Lesions from damage tissue and autoantibodies are directed towards a single
target organ
o Only one organ is being destroyed or damage

AUTOIMMUNE DISEASE ANTIGENS/TARGET ORGANS OR CELLS

-Addison’s disease -Microsomal proteins of adrenal cells (kidney)
-Acute disseminated encephalomyelitis -Basic protein of myelin
-Hashimoto’s thyroiditis -Thyroglobulin, microsomal antigen (thyroid
peroxidase)
-Type 1 DM -islet cells of Langerhans in the pancreas
-Good Pasteur’s syndrome -type IV collagen of basement membrane
-Grave’s disease -TSH receptors
-Myasthenia gravis -Acetylcholine receptors
-Autoimmune chronic active hepatitis -Smooth muscles (anti-smooth antibodies)
-Pernicious anemia -Gastric parietal cell antigens, intrinsic factor
-Sjogren’s syndrome -Salivary gland nucleolar antigens
-Primary biliary cirrhosis -Mitochondria
-Autoimmune myocarditis -Striated cardiac muscle
-Pemphigus vulgaris -Epidermal antigens
-Bullous pemphigoid -Skin basement region antigens
-Autoimmune rheumatic fever -Heart and joint tissue antigen
-Autoimmune glomerulonephritis -Glomerular basement membrane
 Addison’s disease – you have an antibody against microsomal proteins. Anti-microsomal
antibodies.
o Person with this disease may have impaired sodium absorption, because
aldosterone is involved in sodium reabsorption.
- Without aldosterone sodium will not be properly reabsorb in
the kidneys
- Hyponatremia (low sodium in the blood)
o Adrenal cortex produces cortisol and aldosterone.
o Adrenal medulla produces adrenaline or epinephrine and norepinephrine.
 Type 1 DM
o Islet cells of Langerhans in the pancreas are the one that produce insulin particularly the
beta cells of the islets of Langerhans
o Alpha cells of the islet of the Langerhans produces hormone glucagon.
o Delta cells produces somatostatin
 Myasthenia gravis
o Produces anti-acetylcholine receptors; antibody destroying the acetylcholine
- Destroys the receptor on the junction between a nerve cell
and a muscle fiber
o Acetylcholine is a substance responsible for transmitting the impulses from the nerve to
muscle

2. SYSTEMIC OR NON-ORGANIC SPECIFIC

o Lesions from damage tissue and autoantibodies are not confined to any organ.
Sometimes called collagen vascular disease
o Many parts of the body are affected
 Rheumatoid arthritis- chronic, inflammatory joint disease with systemic involvement. Its
hallmark feature is the presence of rheumatoid factors (RF) an antibody reacting with IgG, an
anti-IgG immunoglobin that is produced by B cells and plasma cells in the synovial membrane.

o Can form deformity in the joints (swan neck deformity)

 o Rheumatoid factor is an anti-IgG antibody. It is not exclusive for RA, it can also have SLE
and primary biliary cirrhosis.
o We have a lot of IgG inside the body (most abundant), so if you are producing RF that
have antibody for IgG you are forming a lot of immune complexes inside the body, and
when this immune complex is deposited in the surface of the joint it triggers type 3
sensitivity. The immune complex will trigger compliment activation and as part of the
activation you will form a chemotaxin (C5a)  c5a can attract neutrophil and
phagocytes in the area. Can cause neutrophil to migrate where immune complex is
deposited.  neutrophil will release granules (powerful digestive enzyme) cause
damage to the joint
o Anti-CCP (cyclic citrullinated peptides): most specific antibody for RA
o HLA DR-4 antigen – people with this are more prone with RA (hereditary)
o Uric acid: gout arthritis old age: degenerative joint disease autoimmune:
rheumatoid arthritis
o Bacteria: Neisseria gonorrhoeae
o Limes disease: Borrelia burgdorferi
 Systemic lupus erythematosus – chronic inflammatory multi-organ disorder that predominantly
affects young women of childbearing age.
-HLA DR-2, HLA DR-3 (hereditary)
-Has anti-nuclear antibodies (ANA) -antibodies that react with nucleus. Destroys nucleus of cell.
A group of anti-bodies reacting with the components of nucleus
-most specific antibody Anti-Sm and Anti-ds(double stranded)DNA
o Lupus erythematosus (LE) phenomenon
 It was observed that when peripheral blood from a patient is incubated at 37oC
for 30-60mins, the lymphocytes swell and extrude their nuclear material.
 This nuclear material is opsonized by anti-DNA antibody and complement and is
then phagocytosed by neutrophil. In this case, the cell is now called and LE cells
 LE cells are neutrophil that engulfed nucleus of other cells
 Patient with LE have anti-nuclear antibodies (ANA) – antibodies reacting with
nucleus
 Diagnoses of ANA is using FANA fluorescent anti-nuclear antibody
 Principle: employs IIF (indirect immunofluorescence)
o it uses slide coated with mouse kidney cell or HELA/HEP-2 cells (culture
cells that also contains nucleus)
o then add patient’s serum on the slide then look for ANA (if + then it
means LE)
 (+) The ANA will bind to the HELA cells
 Then wash (to remove excess antibodies)
 Add fluorescent labelled AHG (if there’s an ANA, HELA cell’s
nucleus will fluorest)
o There are four patterns of Fluorescence:
 Homogenous
 Speckled
 Nucleolar
 Centromere
POSITIVE PATTERNS OBSERVED UNDER FLOURESENCE MICROSCOPE:

IMMUNOFLOURESCENT PATTERN ANTIBODY INVOLVED AUTOIMMUNE DISEASE

Homogenous fluorescence Anti-nucleoprotein Rheumatoid arthritis
(staining of the entire nucleus) Chronic active hepatitis
Sjogren’s syndrome
Drug-induced SLE
Myasthenia gravis
MEMBRANOUS/SHAGGY/PERIPHERAL Anti-DNA/native DNA (double SLE
(staining around the nucleus) stranded)
Anti-nucleoprotein (soluble)
SPECKLED FLOURESENCE (numerous Non-DNA nuclear SLE
minute fluorescent points throughout constituents/anti-ENA Scleroderma
the nucleus) (extractable nuclear antigen) Mixed connective tissue
A. Anti-Sm disease
B. Anti-Rnp Sjogren’s syndrome
NUCLEOLAR FLOURESCENCE ONLY Anti-RNA SLE
(fluorescence only on the nucleolus) Scleroderma
FINE SPECKLED PATTERN OF Anti-centromere antibody CREST syndrome (a variant of
FLUORESNCEN SLE)
-calcinosis cutis
-raynaud’s phenomenon
-esophageal dysfunction
-sclerodactyly
-telangiectasia

INTERPRETATION:

HOMOGENOUS (solid, diffuse)

 Whole nucleus fluoresces evenly gold

 Detect antibodies to nDNA, dsDNA, ssDNA, DNP or histones
 High titers are suggestive of SLE
 Low titers are found in SLE, RA, Sjogren’s syndrome, MCTD

PERIPHERAL (Ring, membranous, shaggy, thread)

 Sharp, ring -gold fluorescent of outer edge of nucleus with gradually darkening inner border
blending with a dark nuclear center
 Antibodies to nDNA, dsDNA, DNP
 Seen in active stage of SLE

SPECKLED (mottled)

 Numerous round speckles of green-gold nucleoli of various size against a dark background;
“pepper dots”
 Antibodies to ENA (Sm and RNP)
 Anti-RNP is indicative of other rheumatic disease
NUCLEOLAR

 Multiple, round, smooth green-gold fluorescing nucleoli of various size

 Antibodies to nRNA
 Present in 50% patients with scleroderma, Sjogren’s syndrome and SLE
 Also observed in illness manifesting raynaud’s phenomenon

ANTI-CENTROMERE ANTIBODY (ACA)

 Discrete and speckled pattern

 Antibodies to centromeric chromatin of metaphase and interphase cells
 Highly selective of crest variant and of progressive systemic sclerosis
 Infrequently found in SLE and other MCTD

DIAGNOSES OF AUTOIMMUNE DISEASE:

 ELISA
 INDIRECT IMMUNOFLUORESCENCE (IIF) – FANA
 RIA
 WESTERN BLOT/IMMUNOBLOTTING -gold standard in measuring autoantibodies/ANA
 IMMUNODIFFUSION, BINDING ASSAYS

IMMUNOPROLIFERATIVE DISEASES

-involve malignancies of the immune system

 LEUKEMIAS – malignant cells are present in the bone marrow and peripheral blood and
therefore there would be no one to fight foreign cells (protection)
o Blast cells that proliferate in the bone marrow

+ acute lymphocytic leukemia – characterized by the presence of very poorly differentiated blast cells in
the bone marrow and the peripheral blood. FAB classification includes L1, L2 and L3

+ chronic lymphoid leukemic disorder

o Chronic lymphocytic leukemia/lymphoma – a common hematopoietic

malignancy that involves the expansion of a clone of B cells that have the
appearance of small mature lymphocytes
o Prolymphocytic leukemia – a variant of CLL that is composed of larger
cells with round to oval nuclei and coarser chromatin pattern
o Hairy cell leukemia – a rare, slowly progressive disease characterized by
the infiltration of the bone marrow and spleen by leukemic cells, without
involvement of lymph nodes
  LYMPHOMAS – malignant cells arise in lymphoid tissues such as lymph nodes, tonsils or spleen
o Blast cells that proliferate in the lymphoid organs

+ Hodgkin’s lymphoma – characterized by the presence of reed-Sternberg cells in affected lymph

nodes

o Reed Sternberg cells – large cells with bilobed nucleus and two prominent nucleoli (like an
owl’s eye)

+ non-Hodgkin’s lymphoma

 PLASMA CELL DYSCRASIAS – malignancies that involved the bone marrow, lymphoid organ and
non-lymphoid sites that are not classified as either leukemias or lymphomas

+ multiple myeloma/Kahler’s disease – a malignancy of plasma cells. The most serious and
common of all plasma cell dyscrasias.
- Patients excrete Bence jone protein (antibody like substance,
contains light chain only) in urine

+ waldentrom’s macroglobulinemia/cryoglobulinemia – a malignant proliferation of IgM

producing lymphocytes
HYPERSENSITIVITY

- Heightened state of immune responsiveness. It is an exaggerated response to an innocuous

antigen that results in gross tissue changes that are deleterious to the host.
o Innocuous means harmless but once a person develop hypersensitivity it reacts that’s
why its exaggerated
- One of the disorders of immune system

Description of the Four major hypersensitivity reactions:

-Type 1, 2 and 3 are immediate hypersensitivity because symptoms occur within few minutes to hours
after the exposure to antigen. While type 4 is the delayed type because symptoms may appear after 24
hours or days and sometimes weeks after exposure

-type 1,2,3 is antibody dependent hypersensitivity but in type 4 there is no antibody involved. (antibody
independent)

-type 1 and 2 are compliment dependent while type 1 and 4 are compliment independent

 Type 1 hypersensitivity – an immediate hypersensitivity, sudden allergic responses mediated by

antibodies, primarily IgE. It involves secondary exposure to an offending allergen that bonds to
mast cell-fixed IgE. The antigen crosslink 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. These mediators caused the symptoms of type hypersensitivity
which are manifested within seconds to minute after secondary exposure.
o Atopy – an inherited tendency to respond to naturally occurring inhaled and ingested
allergens with continued production of IgE.
o Allergic rhinitis most common for type 1. It is characterized by rhinorrhea (runny nose).
Treatment is antihistamine, cetirizine

 Type 2 Hypersensitivity – reaction that produce cell damage which is mediated by complement-
fixing antibodies directed against cell surface antigens.

 Type 3 hypersensitivity – also called immune complex mediated hypersensitivity. They are due
to the deposition of Ag-Ab complexes in tissues and blood vessel. These complexes can destroy
the surrounding tissue directly or indirectly by attracting neutrophils to the site of complex
deposition that release that release hydrolytic enzymes, causing local damage.
o Arthur reactions – a necrotic dermal reaction considered to be local immune complex
deposition phenomenon.
o Serum sickness – it results from passive immunization with animal serum, usually horse
or bovine serum used to treat such infections as diphtheria, tetanus and gangrene.

 Type 4 hypersensitivity – a hypersensitivity reaction mediated by sensitized T cells releasing

lymphokines, attracting macrophage to the site and activating them. Once the macrophages
arrive, they begin to cause tissue damage that may develop into a chronic granulomatous
reaction if antigen persists. It is also called delayed type hypersensitivity since following
 secondary exposure to the offending antigen, the manifestation of the interaction does not
appear for more than 24 hrs.

HUMURAL MEDIATED HYPERSENSITIVITY CELL-MEDIATED HYPERSENSITIVITY REACTION

REACTION
 Includes type I, II, III and IV  Include type IV reaction
 Immune reaction is observed minutes  reactions are delayed in time
after antigen exposure  reactions are characterized by significant
 Inflammatory reactions are characterized cell infiltration
by more fluid and erythema (wheal and
flare reaction)

TYPE COMMON DISEASE ANTIGEN (TRIGGER) MEDIATORS

I – Anaphylactic or -Allergic rhinitis, -plant pollen, house -IgE, mast cells
mediated urticaria, hives dust mites, foods (fish,
eggs, chocolate)
-bronchial asthma -fungal or mold spores,
animal danders, animal
hair
-drugs or insect bites -Penicillin, antiseptics,
anesthetics, bee sting
wasp
-tropical eosinophilia -Parasites ( WUCHERERIA
BANCROFTI) - LARVA

-Loeffler’s syndrome -chitin, a protein on

larva of ascaris
lumbricoides
II – Cytotoxic -hemolytic transfusion -antigen on RBC -IgG, IgM, complement
reaction (IgA)
-hemolytic disease of -antigen on RBC
the newborn
-acquired immune -penicillin, quinidine
hemolytic anemia coating RBC
-idiopathic -Ags on surface
thrombocytopenic platelets
purpura
-good Pasteur -Ags on basement
syndrome membrane of the
glomeruli of the kidney
III – Immune Complex -SLE (systemic lupous -nuclear material -IgG, IgM, IgA,
erythematosus) complement
-RA (rheumatoid
 arthritis) -IgG
-Arthus-like reaction
-serum sickness -Aspergillus fumigatus
-horse serum/other
animals
IV – Delayed or -PTB -M. tuberculosis -activated T cytotoxic
Tuberculin -Leprosy -M. leprae cells
-macrophages
-Contact dermatitis -Dyes, metals (nickel), -lymphocytes
cathecol, rubber
V. anti-receptor or -grave’s disease -TSH receptor of acinar -IgG but compliment
stimulator cells fixing
-myasthenia gravis -acetylcholine
VI. Miscellaneous -gram negative -LPS – bacteria
endotoxic shock
-PNH (paroxysmal -Defective RBC
nocturnal membrane
hypersensitivity)

DIAGNOSIS OF HYPERSENSITIVITY REACTION

 Type 1 hypersensitivity
o In-vivo skin test – cutaneous and intradermal testing
o RIST (radioimmunosorbent test) – measures total IgE
o RAST (radioallergosorbent test) – measures allergen specific IgE
 Type 2 hypersensitivity
o DAT (direct antiglobulin test)
 Type 3 hypersensitivity
o Immunoassays and certain agglutination reactions
 Type 4 hypersensitivity
o Tuberculin test/Mantoux test (skin test for TB)
 Injected with PPD (purified protein derivative) and then wait for days to see
result
TYPE 1 HYPERSENSITIVITY

 when an allergen (animal hair ex.) binds to the mediator cell (mast cell or IgE) it will cause an
activation of the IgE and it will bind with the allergen. And the bound allergen to the IgE, will
bind to the mast cell (IgE have a very strong affinity with mast cells and basophil). When an IgE
bounded with allergen are binded to the mast cell it will cause to degranulate (the content will
be release).
o the content contains histamine, ECF-A, neutrophil chemotactic factor and tryptase as
well as newly synthesized mediators such as prostaglandins and leukotrienes and it will
cause the symptoms for type 1 especially histamine.
 Effects of histamine: located inside the muscles and basophil
 Erythema/redness
 Wheal and flare (pantal)
 Increase in mucus production
 Increase in vascular permeability
 Increase in acid production in stomach
 Urticaria
 Increase in smoot muscle contraction
 Prostaglandin – enhance the effect of histamine
 Leukotrienes – resembles the same effect of histamine but are 100x more
potent than histamine.
 The most severe form of type 1 is anaphylaxis (most severe allergy), because it can lead to
multiple organ failure, can cause death.
TYPE 2 HYPERSENSITIVITY

Ex.

 Rh HDN happens between a mother that is RH – and a baby that is Rh +, so it means the red cell
of the baby has Rh antigen (D antigen). And when the mother produces an antibody against the
Rh antigen, the Rh antigen will cross the placenta (it’s possible because it’s basically IgG in
nature)
o Maternal IgG binding, reacting or coating the Rh antigen on the fetal RBC.
o When antibodies of the mother binds with the red cell of the baby it will form immune
complex, it then will trigger compliment activation. Resulting to end result lysis. And
lysis will happen because of a complex called MAC.
TYPE 3 HYPERSENSITIVITY

 When antibodies bind to an antigen it forms immune complex and when the immune complex
deposited in the surface of the cell, the complex triggers the complement activation and the
part of this activation is the formation of chemotaxin (c5a is a complement fragment that acts as
chemotaxin). C5a will cause phagocytes to migrates toward the site where there’s immune
complex
 Most autoimmune diseases are classified in type 3 hypersensitivity
TYPE 4 HYPERSENSITIVITY

 Poison oak can cause contact dermatitis