IMMUNOLOGY & SEROLOGY
3rd Year – 2nd Term
Prelim
Immunology
− Is defined as the study of the molecules, cells,
organs, and systems responsible for the
recognition and disposal of foreign material.
− Began as a branch of microbiology
− The study of infectious disease and the body’s
response to them has a major role for the
development of immunology.
− The concept of germ theory of disease has
contributed to the field of immunology.
*germ theory of disease – Robert Koch
Serology
− Serologic tests are blood tests that look for
antibodies in your blood. They can involve a
number of laboratory techniques.
− Different types of serologic tests are used to
diagnose various disease conditions,
serological test focus on proteins made by your
immune system.
*test on body fluids; antigen, antibodies-protein-
detected by test (IgG etc)
*identify etiological agent
*detecting specific antigen (microorganism, foreign
body)
History of Immunology
Edward Jenner
− First studied the response of the body to foreign
substances
− Observed that dairy maids who had naturally
contracted a mild infection called cowpox
seemed to be protected against smallpox.
*first recorded inducing individual
*Chinese individuals
VARIOLATION- actual virus smallpox
James Phipps
Blossom (cow’s name)
1879
− The first human pathogen, gonococcus, was
isolated by Neisser
1883
− Klebs and Loeffler isolated diphtheria bacilli
which led to the production of the first defined
antigen, diphtheria toxin, by Roux and Yersin in
1888
1888
− The first antibodies against diphtheria, serum
bactericidins, were reported by Nuttal and
Pasteur.
Louis Pasteur
− Father of Immunology
− Observed by chance that older bacterial
cultures would not cause disease in chickens
− Discovered attenuation
*Attenuation- to take a pathogen a less virulent
pathogen and give it to another individual; dead or
lesser virulence that is what similar to the pathogen
(sinovac)
1890
− Von Behring and Kitasato discovered antitoxins
that led to the development of toxoids for
diphtheria and tetanus
1900
− Land Steiner discovered the blood group
antigens and their corresponding antibodies.
This led to the ability to give blood transfusion
without provoking reactions.
*type O – no antigen
1916
− First journal of immunology was published.
Immune System
− “immunity” refers to all mechanisms used by the
body as protection against environment agents
(microorganisms or their products; foods;
chemicals; drugs; pollen; animal hair) that are
foreign to the body
− Arose from the Latin ‘immunis’ meaning
‘exempt’
− Immune system must differentiate between
individual’s own cells and those of harmful
invading organisms
− Must not attack commensal flora that inhabit the
gut, skin, and other tissues to the host’s benefit
*individual resistant to infection
Functions:
− The immune system recognizes and responds
to antigens. In order to protect the individual
effectively against disease, the following tasks
need to be fulfilled.
• IMMUNOLOGICAL RECOGNITION –
recognize a cell as foreign or not
• CONTAIN AND KILL
• TRIGGER IMMUNE
RESPONSE/REGULATION – limits damage to
host cells
• MEMORY-made antibody for a certain antigen
of foreign subs
*progenitor cells – hematopoietic cells/stem cells

IMMUNE SYSTEM
IMMUNITY – resistance, sum total of defenses/defense
mechanism of human body to resist infectious disease
ELI MECHNIKOFF
- Russian Scientist who observe under a
microscope phagocytosis
- Foreign objects introduce to starfish larvae,
unusual cells/amoeboid like cell in the body
who attended to destroy the foreign objects
introduced: SCAVENGER CELLS
INNATE VS ADAPTIVE IMMUNITY
Phagocytosis – process of cells eating cells
*part of immunity/resistance/immune defense
* engulfing
Emil von Behring
• Demonstrated that diphtheria and tetanus toxins,
which are produced by specific microorganisms
as they grow, could be neutralized by the
noncellular portion of the blood of animals
previously exposed to the microorganisms
Almroth Wright
• 1903, linked the two theories by showing that the
immune response involved both cellular and
humoral (fluids or non cell-antibodies) elements.
He observed that certain humoral, or circulating,
factors called opsonins acted to coat bacteria so
that they became more susceptible to ingestion by
phagocytic cells
*antibodies – a type of acute phase reactant:factors in
the body that increase nonspecifically/if there is an
infection
OPSONIZATION – tagging cell as foreign body;
induces phagocytosis
Innate, or natural immunity, is the individual’s ability to
resist infection by means of normally present body
functions. *nonspecific, built in on you since birth
• Nonadaptive or nonspecific
• No prior exposure is required, and the response
lacks memory and specificity
Adaptive, or acquired immunity is a type of resistance
that is characterized by specificity for each individual
pathogen, or microbial agent, and the ability to
remember a prior exposure. *specific immune response
for that specific organism; vaccines
INNATE IMMUNITY
• Is the first line of defense against any infectious
agent
• Several mechanisms are available in the
immunocompetent host, these include:
o Physical and mechanical barrier – skin,
mucous membrane
o Biochemical factors – acids
o Cellular mechanism – readily available
cellular immune system existing in the body
o Role of normal flora
o Inflammatory reactions
PHYSICAL OR MECHANICAL BARRIER
 − Unbroken skin and mucus membrane are
effective mechanical barriers to infectious
agents
− The surface of the skin is also inhibitory to the
growth of most microorganisms because of low
moisture, low pH, and the presence of secreted
inhibitory substance
− Mucus membranes consist of an epithelial layer
and an underlying connective tissue layer

BIOCHEMICAL FACTORS NEUTROPHILS

− Chemical secretions produced by the • Polymorphonuclear neutrophilic (PMN),
body that inhibit microbial growth *and segmented neutrophils, or “segs”
destruction of these cells • Represents approximately 50% to 75% of the
Examples: peripheral WBCs in adults
1. Keratin – skin protein produced by outer most • Round 10 to 15 um in diameter with a nucleus
layer, giving moisture, if dry the skin pores will that has between two and five lobes
open and foreign microorg may enter the skin • Main function of neutrophils is phagocytosis,
2. Hydrochloric Acid resulting in the destruction of foreign particles
3. Bile Salt *increase in bacterial infection
4. Lysozyme – enzyme in body fluids and *azurophilic granules/primary granules- acts as
secretion-tears, help in breakdown of antimicrobial, produces enzyme that is toxic to the
organisms, destruction of bacteria:cell wall microbe
5. Complement *specific granules/secondary granules
6. Interferons – immune response against viruses; * 3-5 lobes
destruction of virally infected cells * certain attraction to chemotactic factors/chemotoxins-
chemicals that causes the cell to migrate to the sight of
infection
CELLS of the Innate Immunity
1. Leukocytes in the Peripheral Blood EOSINOPHILS
a) Neutrophils • 12 to 15 um in diameter and normally make up
b) Basophils between 1% and 3% of the circulating WBCs in
c) Eosinophils a nonallergic person
d) Monocytes
• Increases in an allergic reaction or in response
2. Tissue Cells
to certain parasitic infections
a) Macrophages
• The nucleus is usually bilobed or ellipsoidal and
b) Mast Cells
is often eccentrically located
c) Dendritic Cells
• Eosinophils take up the acid eosin dye and the
cytoplasm is filled with large orange to reddish-
orange granules
• Capable of phagocytosis but are much less
efficient than neutrophils
• Eosinophils can neutralize basophil and mast
cell products, the most important role of
eosinophils is regulation of the immune
response, including of mast cell function.
*granulitic; cationic proteins-kill the large parasitic
worms
BASOPHILS
• The least numerous of WBCs found in
peripheral blood, representing less than 1% of
all circulating WBCs.
• The smallest of the granulocytes, basophils are
slightly larger than RBCs (between 10 to 15 um
in diameter)
• Contain coarse, densely staining deep-bluish-
purple granules that often obscure the nucleus
• Granules include histamine, cytokines, growth
factors, and a small amount of heparin, all of
which have an important function in inducing
and maintaining allergic reactions
* antibody mediated autoimmune diseases
*histamine-contracts smooth muscle in allergic
reactions; neutralize the granules released by basophils
* help in regulation of the T helper cells
*stimulate B cells to produce anitbodies-IgE-specific
Antibody specific for allergic reaction
MONOCYTES
• Largest cells in the peripheral blood with a
diameter that can vary from 12 to 22 um (the
average is 18 um)
• Has an irregularly folded or horseshoe-shaped
nucleus that occupies almost one-half of the
entire cell’s volume
• The abundant cytoplasm stains a dull grayish
blue and has a ground-glass appearance
because of the presence of fine dust like
granules. Digestive vacuoles may also be
observed in the cytoplasm
• Monocytes make up between 4% to 10% of
total circulating WBCs, stay in peripheral blood
for up to 30 hours; they then migrate to the
tissues and become known as macrophages
• Known as macrophage precursors
*agranulocytes
MACROPHAGES
• Arise from monocytes, unlike monocytes,
macrophages contain no peroxidase
• Play an important role in initiating and
regulating both innate and adaptive immune
responses
• Specific Names:
1. Alveolar macrophages (lungs)
2. Kupfer cells (liver)
3. Microglial cells (brain)
4. Osteoclast (bone)
5. Histiocytes (connective tissue)
MAST CELLS
• Resemble basophils
• Are distributed throughout the body in a wide
variety of tissues such as skin, connective
tissue, and the mucosal epithelial tissue of the
respiratory, genitourinary, and digestive tracts
• Life span of between 9 to 18 months
• Play a role in allergic reactions, can also
function as antigen-presenting cells (APCs)
*gets the soluble debris<after phago> to
present to the specific immune cell to identify
what specific antibody to produce
• Can both enhance and suppress the adaptive
immune responses
*has acid phosphatase, alkali phosphatase, protease
DENDRITIC CELLS
• Covered with long membranous extensions that
make them resemble nerve cell dendrites
• Discovered by Steinman and Cohn in 1973
• Seen in lymphoid and nonlymphoid tissues
• Considered the most effective APC
• Most potent phagocytic cell
*from bone marrow, not same lineage to WBC
*present this debris to T cell, initiate adaptive immune
response, same with macrophages
ADAPTIVE IMMUNITY
• Also known as the specific immune response or
Acquired immunity
• Is a defense system that protects the body
against pathogenic microorganisms and other
type of disease such as cancer
• It allows the body to recognize, remember, and
respond to a specific stimulus, an antigen
Passive Immunity – the immunity in which antibodies
produced elsewhere and are given to an individual, ex.
vaccination
1. Naturally acquired passive immunity –
infected mother then antibodies produced is
passed through the baby; acquired immunity
2. Artificially acquired passive immunity –
given to an indi
Active Immunity – your immune system will work itself
for immunity
1. Naturally acquired active immunity –
encountered in daily life
2. Artificially acquired active immunity – the
part or dead organism is given directly to host;
ex. Cowpox- less virulence strains. Your body
make antibodies against it
LYMPHOCYTES
• Represent between 20% and 40% of the
circulating WBCs
• The typical small lymphocyte is similar in size to
RBCs (7-10 um in diameter) and has a large
rounded nucleus that may be somewhat
indented.
• These cells are unique because they arise from
an HSC and then are further differentiated in the
primary lymphoid organs, namely the bone
marrow and the thymus
*second most abundant;
 • 3 Major Populations
1. T CELLS (Helper, Cytotoxic
Regulatory) – differentiated, made by BM;
produces cytokines that produces
immunity by stimulating the B cells to
produce antibodies and kill target cells,
trigger adaptive immunity and destroy
abnormal cells like cancer cells/tumor
cells
*helper – help destroy and identify; help
activate immune respone
*cytotoxic – has the specific function for
destroying; toxic to antigen or microbe
*regulatory – uy tama na t-cell
* to identify these 3, CB markers will
help
2. B CELLS – derived from lymphoid
precursors but differentiated to BM;
produces antibodies and memory cells;
produces antibody against the specific
antigen
3. NK CELLS – does not have a marker
present in B and T cells; just to kill target
cells without prior exposure to them; do
not require thymus to mature but appear
to the BM
FACTORS ASSOCIATED WITH IMMUNOLOGIC
DISEASES
Age
• Non-specific and specific body defense are
present in the unborn and newborn infants, many
of these defenses are not completely developed
• Older adult’s certain natural barriers to infection
break down such as changes in the skin and in the
lung weakening of specialized defenses against
foreign invasion including the cough reflex
Nutrition
• Good nutrition is known to be important to
growth and development
Genetic Factor
• Genes linked immune disorders
++ Body Temp, Oxygen Tension, Hormonal
Balance and PH
LYMPHOID ORGANS
&
LYMPHOID TISSUE
• The immune system is a network of cells and
organs that extend through out the body and
function as a defense against infection
• In mammalian immunologic development, the
precursors of lymphocytes arise from
progenitor cells of the yolk sac and liver
• Later in the fetal development the bone marrow
becomes the main provider of undifferentiated
progenitor cells
PRIMARY LYMPHOID TISSUE
Bone Marrow
• Considered one of the largest tissues in the
body and it fills the core of all long flat bones
• The main source of hematopoietic stem cells
• The bone marrow also plays a role in the
differentiation of progenitor cells into B-
lymphocytes and functions as the burse
equivalent in human
*Bursa of Fabricus/Fabricius- primary lymphoid organ of
the birds; humans have the same bursa where B-
lymphocyte is derived from
Thymus
• A gland situated in front of the heart and behind
the sternum
• Progenitor cells that leave the bone marrow
migrate to the thymus for proliferation and
differentiation that is facilitated by a hormone,
thymosin
• Thymus diminishes in size as humans age, but
it is still capable of producing mature T
lymphocytes, although at a diminished rate
*has hormone thymosin-stimulates the
development/maturation of T-cell
SECONDARY LYMPHOID ORGAN
• The function of the secondary lymphoid organs
is to maximize encounters between
lymphocytes and foreign substances, and it is
from this size that most immune responses are
launched
• Organs
1. Lymph nodes
2. Spleen
3. Gut-associated lymphoid tissue
4. Tonsils
5. Blood and others

Lymph Node
• Surrounded by a capsule
• Outer cortex contains collection of B cells,
macrophages and follicular dendritic cells
*found specifically in the primary follicle
• T cells are found in the paracortex
• Medulla contains some T cells, B cells,
macrophages and plasma cells *produce anti
bodies and memory cells-plasma cells
*acts as a lymphoid filter of the lymphatic system
*after lymphocytes mature, they go to lymph nodes
where the fighting of infection happened (lusay)

OTHER SECONDARY ORGANS

• Mucosal – associated lymphoid tissue (MALT)
– found in the gastrointestinal, respiratory and
urogenital tract
• Cutaneous – associated lymphoid tissue
(CALT) – found in skin and epidermis

LINE OF DEFENSES
• First line of defense/ primary (non-
specific)
− Intact skin, ciliated epithelium in
airways, acids, body fluids (tears and
saliva)
• Second Line of defense/ secondary
(non-specific)
− Mucosal surfaces, native bacterial flora/
microbial flora, inflammation, fever,
phagocytosis, NK cells and
macrophages
• Tertiary line of defense/tertiary
(specific)
− B and T cells, and Antibodies
MIDTERMS
MAJOR HISTOCOMPATIBILITY COMPLEX
The ability of our immune stem to recognize its own cells
and distinguish those cells from foreign pathogens
depends on PROTEIN MARKERS (MHC) found on cell
membranes.
*important function of leukocytes, immune cells
*differentiate healthy cells and not
MHC is protein in all animals
In humans – HLA: human leukocyte antigen
1. Class II MHC binds in variant chain to block binding
of endogenous antigen.
2. MHC complex goes through Golgi complex.
3. Invariant chain is degraded, leaving CLIP fragment.
4. Exogenous antigen taken in and degraded and
routed to intracellular vesicle.
5. CLIP fragment exchanged for antigenic peptide.
6. Class II MHC antigenic peptide is transported to cell
surface.
7. Class II MHC peptide complex binds to CD4+ T cell.

*macrophages – produces interleukin (1)→ recruiting

MCH Class I Gene T helper cells-will now call other immune cells
• Found on MOST nucleated cells
• Used to differentiate healthy hist cells
from infected cells (virus or bacteria infected MHC Class III Gene
cells) • Encode for secreted proteins that have immune
• It is recognized by only T cytotoxic functions (Complement proteins &
cells inflammatory molecules/molecules
*has 4 polypeptide units involved in inflammation)
Alpha 1, a2, a3, beta-microglobulin • DO NOT play a role on presentation of
ALPHA 3 – only one connected to the cell antigenic peptides
membrane/plasma membrane
• MHC I not so stable coz only 1 peptide is
connected
*recognition
If healthy cell – it will produce self protein, to not be
destroyed, MHC will show that it is a healthy cell so it
won’t phagocyte that cell

If virus containing cell – it will show the protein part of

the virus → lymphocyte or nucleated cell will now
identify it as none-self then trigger cellular immune self
and undergo apoptosis

1. Endogenous antigen within cytosol is degraded by

proteasome.
2. Peptides transported into endoplasmic reticulum by
TAP.
3. Alpha chain of class I MHC binds B2-microglobulin.
4. Alpha chain of class I MHC binds peptide.
5. Peptide-class I MHC transported to Golgi complex
and then to cell surface.
6. Class I MHC peptide binds to CD8 + T cell.

MHC Class II Gene

• Found on specific immune cells such
as B-lymphocytes, macrophages, dendritic
cells, and some T-lymphocytes
• Functions in helping immune cells
communicate with one another
• Recognized by only T-helper cells
*does not identify self or none self; calls immune cell to
communicate with one another
*composed of HOMOLOGOUS CHAINS OF BETA AND
ALPHA
*a1, a2, b1,b2
*Both Alpha 2 and Beta 2 connects to the cell
membrane
 COMPLEMENT
• A complex (proteins) series of more than 30
proteins that play a major part in amplifying the
inflammatory response to destroy and clear foreign
antigens.
• Serve as an important link between innate and
adaptive immunity
• Recognizes cellular debris such as apoptotic cells
and immune complexes, tagging them from removal
innate immune cells.
*Soluble, cell-bound proteins which if activated will:
Lyse foreign cell, opsonized (buslotan ang cell) big
cells, tag the invaders, so the immune cell can clear
these specific antigen
*WHAT ACTIVATES OUR COMPLEMENT SYSTEM?
Proinflammatory responses/ ANTIGENS in general
Once activated: increase vascular permeability, recruit
immune cells (neutron, mono, lympho), trigger adaptive
immune response
*Can give: tissue damage (apoptosis, or destruction to
oponization), inflammation
*IF DEFICIENCY IN COMPLEMENT: individual is
susceptible to infections
*IF OVER/INCREASES COMPLEMENT: autoimmune
disease/disorders
Complement System: Pathways
Activation of complement system can be initiated by
either an antigen antibody complex or by variety of
foreign surfaces
*Functions: opsonic function, inflammatory function
(where after activation of complement system will now
cause the induction of histamine release stimulation of
inflammatory response, cytotoxic function (final stage
in complement cascade)
*KNOWN AS THE RECOGNITION UNIT: Component
of complement unit (C1rqs or C1 unit)
*KNOWN AS 1ST MEMBRANE ATTACK UNIT: C5
*COMPONENT OF COMPLEMENT THAT IS KNOWN
AS THE FINAL MEMBRANE ATTACK UNIT: C8 & C9
3 different pathways:
1. Classical Pathway
2. Alternative Pathway
3. Lectin Pathway
CLASSICAL PATHWAY
• Involves 9 proteins that are triggered
primarily by antigen-antibody combination.
• The immunoglobulin classes that can
activate the classical pathway include IgM,
IgG1, IgG2, and IgG3, but NOT IgG4, IgA, or
IgE.
*most efficient immunoglobulin that can activate your
complement pathway: IgM – has multiple binding sites,
takes only 1 molecule attached to adjacent antigenic
determinants to initiate classical pathway
*among IgG which is most effective to initiate classical
pathway: IgG3 next IgG1, IgG2 no IgG4
• C-reactive protein (CRP), several
viruses, mycoplasmas, some protozoa, and
certain gram-negative bacteria such as
Escherichia coli CAN DIRECTLY INITIATE
THIS PATHWAY (but commonly initiates
alternative and lectin pathway not classical).
STAGES OF ACTIVATION
1. Recognition
2. Activation
3. Membrane Attack Complex (MAC)
1. RECOGNITION
*C1qrs unit binds to the Fc portion of 2 antibody
molecule and will now have domino effect to the
classical pathway
• C1 - has 3 subunits (C1q, C1r, C1s) =
NEEDS Ca2+
• C1q binds to Ab
• C1r & C1s (serine protease enzymes =
ZYMOGENS) which generate enzyme activity
to begin cascade.
• C1s is activated and cleaves C4 and
C2 to form C4b2a, also known as C3
convertase (activate C3)
2. ACTIVATION *goal is to activate C5 convertase
• Combination of C4b and C2a is known as C3
convertase (NOT STABLE)
• C3 convertase binds to C3b subunit LEADING
to C4b2a3b (C5 convertase)
• The cleavage of C3 convertase to C3b
represents the most significant step in the entire
process of complement activation.
*What will happen to subunit C3a – remain unbound;
increasing vascular permeability, inflammatory
response
*C4a? – released to into the fluid phase; unbound;
inflammatory functions
*among the proteins of classical pathway, which is also
known as C1 esterase? – C1s
3. MEMBRANE ATTACK COMPLEX
• C5 convertase (C4b2b3b) splits C5a that is
released into circulation, whereas C5b attaches
to the cell membrane, forming the beginning of
the MAC
• The complex of C5b-C6-C7-C8 and C9 is
known as C5b-9 or MAC
• C9 speeds up lysis of target cell
*in general MAC – will attack the antigen
*other name of alternative pathway – properdin
pathway
ALTERNATIVE PATHWAY
• First described by Pillemer and his associates
in the early 1950s
• Originally names for the protein properdin
• Can be activated on its own
• Triggering substances:
1. Bacterial cell walls, especially those
containing lipopolysaccharide
2. Fungal cell walls
3. Yeast
4. Viruses
5. And virally infected cells
6. Tumor cell lines, and
7. Some parasites, especially
trypanosome (no protozoa)
*DO NOT NEED RECOGNITION UNIT – C1, C4, C2
*because C3 is activated by another route by
lipopolysaccharide or endotoxin from the cell wall of
bacteria, fungi
• 1ST STEP – conversion of C3
• C3bBb (C3 convertase of the alternative
pathway) (B= factor b; b= factor D)
• C3bBbP3b (C5 convertase of the alternative
pathway) (P=properdin to stabilize; substitute to
C4b2a in classical pathway)
• After C5 is cleaved, the pathway is identical
to the classical pathway
*to activate C3- hydrolyzed by water to produced C3b,
now bind with factor B→then B will bound with Factor
D → will now bind with C3b with the help of
PROPERDIN(free flowing in the plasma) to stabilize
C3b
LECTIN PATHWAY
• Antibody independent and has similarities
with classical pathway
• Most recently described of the three activation
pathways of complement, it is probably the
most ancient.
• Activated by recognition of surface moieties
that are found on pathogens
• Provides an additional link between the innate
and acquired immune response
*structurally similar with classical pathway
*share components of C4 and C2
*differs in RECOGNITION UNIT (C1q,r,s)
Components:
• MBL: Mannose-binding Lectin
• MASP-1 (C1r of classical) *Mannose-binding
Lectin Associated Protease
• MASP-2 (C1s of classical)
• ALL 3 components form complex on
PATHOGEN MEMBRANE (like C1qrs of
classical pathway) *attracted to lectin
(mannose) of the organism
*these attached to the lectin
CLASSICAL:
• C1qrs→C4b2a(C3convertase)→C4b2
a3b(C5convertase)-MAC
ALTERNATIVE:
• C3bBbP(C3convertase)→C3bBbP3b(
C5convertase)→MAC
LECTIN:
• MBL-MASP1&MASP2→C4b2a
(C3convertase)→ACTIVATION-MAC
Regulation of Complement System Activation
CLASSICAL PATHWAY regulation or inhibition
1. C1 inhibitor – found in the blood/circulation
• Deficiency – Hereditary Angioedema
*binds to the Fc region of the antibody if no antigen is
bound to the Fab region of antibody
*inhibit recognition process
ALTERNATIVE PATHWAY regulation or inhibition
1. Factor H – found in the blood
2. Factor I – found in the blood
*factor H is technically cofactor with Factor I –
inactivate C3b and C4b
OTHER REGULATORS: (General Complement
System)
1. Decay accelerating factor (DAF) & CD59 – proteins
which protects own/self cells from complement
• Deficiency – PNH (brownish urine is
observed due to overlysed RBCs)
*Paroxysmal Nocturnal Hemoglobinuria
*inside the own cell, owned by own cell
*DAF – inhibits C3 convertase
*CD59 – inhibits membrane attack complex
Complement & Disease States
Major pathway component Deficiencies:
• C2 – most common
• MBL – 2nd most common & associated
with pneumonia, sepsis, Neisseria infection,
cancer, & SLE.
• C3 – most serious & associated to
severe recurrent infections and
glomerulonephritis
Regulatory factor Deficiencies:
• Decay Accelerating Factor (DAF) – PNH
- DAF deficiency is associated with
CD59 (MIRL)
 • C1-INH – Hereditary Angioedema
Types:
1. Type 1: Low C1-INH(ibitor)
2. Type 2: normal level of C1-INH
nonfunctional
• Genetic mutations of Factor H, MCP, Factor I,
Factor B, and thrombomodulin, as well as
autoantibodies to Factor H and Factor I can lead
to HUS (Hemolytic Uremic Syndrome *E. coli,
O157H7)
RED CELL SUSPENSION
Materials Needed
• Adapter
• Applicator Stocks
• Beakers (250mL capacity) u Centrifuge
machine
• Evacuated tubes (EDTA) u Forceps
• Rubber bulbs
• Serological pipettes (0.1, 1.0, 2.0, and 5.0 mL)
• Test tubes (5mL capacity)
• Test tube brush
• Test tube rack
• Two-way needle
• Wash bottles
Reagents:
• Distilled Water
• Normal Saline Solution (NSS)
Sample:
• EDTA Blood Sample
ABSTRACTION
Red Blood Cell suspension is an important
laboratory made or manufacturer readily made reagent
antigen in the Blood Bank Laboratory. This ranges from
3-5% (commonly 2-5%) concentration of Red Blood
Cells suspended in a specific volume of physiologic
saline. Every hemagglutination reaction must happen
using a 3-5% RBC suspension. This concentration of
RBC is the ideal which hemagglutination reaction
occurs. Heavy suspension, greater than 5%
concentration, means high antigen concentration. This
may result into post zone phenomenon where there is
an excess of antigen concentration. This may result into
prozone phenomenon where there is excess antibody
and insufficient antigen concentration leading to a false-
negative result. Prozone remedy is simple with use of
dilution.
PROCEDURE
a. Washing of Red Cells
b. Preparation of Red Cell Suspension (Accurate
Method)
Washing of Red Cells
*remove plasma, microaggregates, cytokines,
unwanted antibodies that will interfere with Antigen and
but Antibody reaction
1. Spin the EDTA Blood Sample to separate plasma
from the red cell.
2. Transfer 2 drops of blood using a Pasteur pipettes
from the Packed Red Cell to a tube labeled with “Px
WRC” (Patient Washed Red Cells)
3. Fill the tube into ¾ using a physiological saline.
4. Cover with Parafilm and mix gently yet thoroughly
by inverting
5. Spin for 1 min. at 3,400 rpm
6. Remove the Parafilm. Discard the saline by
immediately and blot in a tissue paper to remove
excess saline.
7. Repeat procedure 3-6 for a minimum of 2 times. (A
total of 3 times minimum washing step) *3-4 times,
minimum of 3)
Preparation of RED CELL SUSPENSION (Accurate
method)
1. Make 3, 4, and 5% RED CELL SUSPENSION
2. RBC suspension is made by using the formula:
3. %RCS= Solute(PVC) / Solvent (Volume of NSS)
+ Soluble Volume) x 100
4. Get another clean tube and label again as “Px
RCS” (Patient Red Cell Suspension)
5. Decide first for the desired total volume of
suspension and the concentration
6. Derive the formula above to get the exact volume
of solute needed to make the desired
concentration and volume of RCS
7. Use the derived formula below:
8. PCV = %RCS x NSS Volume / 100
9. Once the PCV has been solved, transfer the
correct PCV, as computed, into the labelled clean
tube.
10. Add the exact volume of diluent (NSS)
11. Cover with Parafilm and mix gently by inversion.
 HYPERSENSITIVITY
• Hypersensitivity (allergy) is an inappropriate
immune response that may develop in the humoral
or cell-mediated responses
• Was first termed anaphylaxis
• Can be systematic, which often leads to shock and
can be fatal, or localized, which is various atopic
reactions
TYPES OF REACTIONS
There are four types of reactions:
Type I – IgE mediated
Type II – Antibody-Mediated (IgG or IgM mediated)
Type III – Immune Complex-Mediated (IgG, IgM)
Type IV – Delayed-Type Hypersensitivity (DTH)
TYPE I: IgE-MEDIATED HYPERSENSITIVITY
*related to anaphylaxis reactions
• IgE-mediated – “key reactant”
• Immediate hypersensitivity (minutes)
• Most allergic reactions
• Antigens that are trigger formation of IgE are called
atopic antigens, or allergens
• Allergic reaction – due to a genetic predisposition
(kulang na genetic codes, and if met by the body
and think it is anitgen)
• 2 steps:
o Sensitization (initial exposure)
o Activation (subsequent exposure)
*immunogen – all immunogen are antigens; can trigger
immune response
*antigen – not all; can trigger or cannot trigger immune
response; foreign materials not normal in the body
Triggering of Type I by IgE
• IgE normal levels is 150 ng/mL
• IgE is regulated by Th2 cells
• Th2 produces interleukin-3 (IL-3), interleukin-4 (IL-
4), interleukin-5 (IL-5), interleukin-9 (IL-9), and
interleukin-13 (IL-13) in people with allergies.
• Allergens
− Foods
− Molds
− Drugs/meds
− Pollen
− Skin contact (latex, lotions and soaps)
How is a Type I hypersensitive response different from
a normal humoral response?
• The plasma cell lineage of the B cell that
exogenously processed the allergen secretes IgE,
not any of the other isotypes.
*humoral immune response – used antibody in
response to allergen
*cell-mediated – uses cell components like neutrophils
MAST CELLS AND BASOPHILS
Mast Cells
• Principle effector cells of immediate hypersensitivity
• Derived from precursors in the bone marrow that
migrate to specific tissue sites to mature
• Most prominent in connective tissue, the skin, the
upper and lower respiratory tract, and the
gastrointestinal tract
• Contains numerous cytoplasmic granules (e.g.
Histamine)
• They release a variety of cytokines and other
mediators that enhance he allergic response
Basophils
• Represents approximately 1 percent of the white
blood cells in peripheral blood
• Half-life of about 3 days
• Contain histamine-rich granules and high-affinity
receptors for IgE, just as in mast cells
• Respond to chemotactic stimulation and tend to
accumulate in the tissues during an inflammatory
reaction
The chemically active effectors within the granules
released via degranulation are called mediators. This
group includes:
• Histamines
• Leukotrienes
• Prostaglandins
• Cytokines
Sensitization – mast cells
Activation – mast cells, and basophils
What is the sequence of events in an IgE-mediated
hypersensitive response?
1. The plasma cells secrete IgE.
2. These IgE bind to Fc receptors on sensitized
mast cells and blood basophils.
3. When the allergen appears again (usually a few
weeks after the first exposure), it cross-links the
mIgEs (memory) and causes degranulation,
releasing granules.
4. Mediators within these granules act on the
surrounding tissues such as smooth muscle,
small blood vessels, and mucous glands.
Type I Hypersensitivity
Mild Symptoms
• Hives (urticaria)
• Eczema
• Allergic Rhinitis
• Asthma
Severe Symptoms
• Increase vascular permeability
• Airway constriction
• Anaphylactic shock
And just look at all these popular allergens!
What is available in the Western medicine arsenal
of drugs?
• Antihistamines – block the binding of
histamine on target cells (corticosteroids and
epinephrine – needs doctors consent,
diagnosed; BEES- can trigger anaphylactic
shock- given epinephrine)
• Immunotherapy – treat the patient with
increased doses of the allergen
(hyposensitization) to reduce severity of the
response
• Or, practice better dust control, find another
home for Fido, and don’t eat those strawberries!
TYPE II: ANTIBODY-MEDIATED CYTOTOXIC
HYPERSENSITIVITY/cytolytic hypersensitivity
• IgM & IgG mediated
• Involves the antibody mediated destruction of
cells
• Can mediated cell destruction by activating the
complement system to create pores in the
membrane of the foreign cell
• Can also mediated by Antibody-Dependent Cell-
Mediated Cytotoxicity (ADCC) where the Fc
receptors bind to Fc receptor of antibody on the
target cell and promote killing *antibody is the
reason why the cell lyse
*target specific antigen; can destruct cells- e.g. drug
penicillin: can be seen in blood system and attaches to
RBC- can trigger type 2 lysing the cell including antigen;
complement system activation; drug induced hemolytic
anemia
*non-cytotoxic/cytotoxic mechanism – antibody
mediated cellular destruction/disfunction; blocking of
receptors causing over activation of cell with antibody
producing a lot of interleukins and etc; Myostania
Gravious and Grave’s disease
TYPE II HYPERSENSITIVITY: DISEASE
1. Transfusion Reactions
2. Hemolytic Disease of the Newborn
• Erythroblastosis fetalis
3. Type 2 reactions involving tissue antigens
4. Hemolytic anemia
• Autoimmune
• Drug-induced
Transfusion reactions:
• Antibodies of the A, B, and O antigen are usually of
the IgM class (these antigens are called
isohemagglutinins)
• For example an A individual produce
isohemagglutinins to B-like epitopes but not to A
epitopes because they are self
• Person who are transfused with the wrong blood
type will produce anti-hemagglutinins causing
complement mediated lysis
• Antibodies are usually of the IgG class
• Transfusion reactions can be delayed or immediate
but have different Ig isohemagglutinins
• Immediate reactions has a complement-mediated
lysis triggered by IgM isohemagglutinins
• Delayed reactions induce clonal selection and the
production of IgG which is less effective in activating
the complement
• This leads to incomplete complement-mediated
lysis
• Cross-matching can detect antibodies in the sera to
prevent this
Hemolytic Disease of the Newborn
• This is where maternal IgG antibodies specific for
fetal blood group antigens cross the placenta and
destroy fetal RBC’s
*first stage of pregnancy
• Erythroblastosis fetalis – severe hemolytic disease
of newborns
o Most commonly develops when an Rh+ fetus
expresses an Rh antigen on it’s blood that and
Rh- mother doesn’t recognize
Drug-Induced Hemolytic Anemia
• This is where certain antibiotics can absorb
nonspecifically to the proteins on RBC
membranes
• Examples: penicillin, streptomycin
• Sometimes antibodies form inducing
complement-mediated lysis and this
progressive anemia
• When drug is withdrawn the hemolytic anemia
disappears
TESTING:
Direct Coombs Test
• Px RBC is separated + coombs reagent
(composed of antibodies)
o Agglutination = there is antibodies present in
the RBC surface *antibody-antibody reaction
Indirect Coombs Test
• For blood group incompatibility
• Px Serum + RBC’s with known Ag + coombs
reagent
o Agglutination = there is antibodies or
complement in the serum
*detects antibody before exposure to antigen

TYPE III – IMMUNE COMPLEX-MEDIATED
HYPERSENSITIVITY
*soluble antigen
*antigen is free flowing not attached to cells
*common to insect bites
*same with type 2- trigger complement system
• Reaction with antibodies create immune complexes
• These generally facilitate the clearance of antigen
by phagocytosis
• Large amounts of immune complexes can lead to
tissue damage (Type III reaction)
• The magnitude depends on the quantity of immune
complexes and their distribution
• The complexes get deposited in tissues:
✓ Localized reaction is when they are
deposited near the site of antigen entry
✓ When formed in the blood reaction can
develop wherever they are deposited
• Deposition of these complexes initiates a reaction
that results in the recruitment of neutrophils
• Tissue is injured by the granular release from the
neutrophil (attempted phagocytosis release lytic
enzymes that cause the damage)
Generalized Type III Reactions:
✓ Large amounts of antigens enter the blood
stream and bind to antibody, circulation
immune complexes can form
✓ These can’t be cleared by phagocytosis and
can cause tissue damaging Type III reactions
Serum Sickness – type III hypersensitivity reaction that
develops when antigen is intravenously administered
resulting in the formation of large amounts antigen-
antibody complexes and the deposition in tissue
Other conditions caused by Type III –
1. Infectious Diseases
• Meningitis
• Hepatitis
• Mononucleosis
2. Drug Reactions
• Allergies to penicillin and sulfonamides
3. Autoimmune Diseases
• Systematic lupus erythematosus (SLE)
• Rheumatoid arthritis
Arthur’s Reaction – immune complex
reaction/hypersensitivity
Serum Sickness Reaction
Infections like SLE
Good Pastures
Rheumatic Fever
Rheumatoid Arthritis
Occupational Disease - Farmer’s Lung
Pigeons Reader’s Disease
TYPE IV HYPERSENSITIVITY
• a.k.a cell mediated hypersensitivity or delayed
type hypersensitivity
• T-cell mediated (t-helper, t-cytotoxic)
• First described in 1890 by Robert Koch
What is delayed type hypersensitivity (DTH)?
• A hypersensitive response mediated by
sensitized TDTH cells, which release various
cytokines and chemokines
• Generally, occurs 2-3 days after TDTH cells
interact with antigen
• An important part of host defense against
intracellular parasites and bacteria
Phases of the DTH Response
1. Sensitization phase: occurs 1-2 weeks after primary
contact with Ag
What happens during this phase?
• TH cells are activated and clonally expanded by
Ag presented together with class II MHC on an
appropriate APC, such as macrophages or
Langerhan cell (dendritic epidermal cell)
• Generally, CD4+ cells of the TH1 subtypes are
activated during sensitization and designated
as TDTH cells
2. Effector phase: occurs upon subsequent exposure
to the Ag
What happens during this phase?
• TDTH cells secrete a variety of cytokines and
chemokines, which recruit and activate
macrophages
• Macrophage activation promotes phagocytic
activity and increased concentration of lytic
enzymes for more effective killing
• Activated macrophage are also more effective
in presenting Ag and function as the primary
effector cell
Types of Reactions
Protective Role of DTH Response
Intracellular Intracellular Contact
bacteria viruses Antigens
Mycobacterium Herpes simplex Hair dyes
tuberculosis virus
Mycobacterium Measles virus Poison ivy
leprae
Detrimental Effects of DTH Response
• The initial response of the DTH is nonspecific
and of the results on significant damage to
healthy tissue
• In some cases, a DTH response can cause
such extensive tissue damage that the
response itself is pathogenic
 • Example: Mycobacterium tuberculosis – an
accumulation of activated macrophages whose
lysosomal enzymes destroy healthy lung tissue
✓ In this case, tissue damage far
outweighs any beneficial effects

How Important is the DTH Response?

• AIDS illustrates the vitally important role of
the DTH response in protecting against
various intracellular pathogens.
• The disease cause severe depletion of CD4+
T cells, which results in a loss of the DTH
response.
• AIDS patients develop life-threatening
infections from intracellular pathogens that
normally would not occur in individuals with
intact DTH responses.
FINALS
IMMUNODEFICIENCY DISORDERS
Immunodeficiencies are disorders in which a part of the
body’s immune system is missing or dysfunctional.
People with these conditions have a decreased ability
to defend themselves against infectious organisms and
are more susceptible to developing certain types of
cancer.
Immunodeficiencies can be inherited or acquired
secondary to other conditions such as certain infections,
malignancies, autoimmune disorders, and
immunosuppressive therapies.
Secondary Immunodeficiency is the acquired
immunodeficiency syndrome (AIDS), which is caused
by the human immunodeficiency virus (HIV).
Primary Immunodeficiencies (PIDs), which are
inherited dysfunctions of the immune system. Several of
the most important immunodeficiency syndromes show
X-linked inheritance and, therefore, affect primarily
males.
Defects in humoral immunity (antibody production)
result in pyogenic (i.e., pus-forming) bacterial infections,
particularly of the upper and lower respiratory tract.
Recurrent sinusitis and otitis media (i.e., ear infections)
are common.
Agammaglobulinemia, conditions in which antibody
levels in the blood are significantly decreased.
RHEUMATOID ARTHRITIS
Causing chronic inflammation in joints and particular
tissues
LAB TESTS
Sheep Cell Agglutination Test
Latex Fixation Test
Sensitized Alligator Erythrocytes Test
Bentonite Flocculation Test
RA is another example of a systemic autoimmune
disorder. It affects about 0.5% to 1.0% of the adult
population, but prevalence varies with ethnicity and
geographic location. Typically, it strikes individuals
between the ages of 25 and 55. Women are three times
as likely to be affected as men.
RA can be characterized as a chronic, symmetric, and
erosive arthritis of the peripheral joints that can also
affect multiple organs such as the heart and the lungs.
The strongest environmental risk factor for RA is
believed to be cigarette smoking, which doubles the risk
of developing the disease.
RA is caused by an inflammatory process that results in
the destruction of bone and cartilage. The lesions in
rheumatoid joints show an increase in cells lining the
synovial membrane and formation of a pannus, a sheet
of inflammatory granulation tissue that grows into the
joint space and invades the cartilage. Infiltration of the
inflamed synovium with T and B lymphocytes, plasma
cells, dendritic cells, mast cells, and granulocytes is
evidence of immunologic activity within the joint.
HASHIMOTO’S DISEASE
Destruction of thyroid gland; organ specific
ANTIGENS PRODUCES
Thyroglobulin
Microsomal Antigen (ANTIGEN)
Anti-thyroid peroxidase (anti-microsomal antibodies)
Anti-thyroglobulin antibodies (ANTIBODIES)
Hashimoto’s thyroiditis, also known as chronic
lymphocytic thyroiditis, was discovered in Japan in 1912
by Dr. Hakaru Hashimoto. It is now considered to be the
most common autoimmune disease, affecting about 8
out of every 1,000 individuals.89 The disease is most
often seen in middle-aged women; in addition, women
are 5 to 10 times more likely to develop the disease than
men.
Patients develop an enlarged thyroid called a goiter,
which is irregular and rubbery. Patients also produce
thyroid-specific autoantibodies and cytotoxic T cells.
Symptoms of hypothyroidism include dry skin,
decreased sweating, puffy face with edematous eyelids,
pallor with a yellow tinge, weight gain, fatigue, and dry
and brittle hair.
GRAVE’S DISEASE
- Unregulated secretion of thyroid hormones T3
and T4
- TSH-receptor like → produces Anti-TSH
Antibody
• Graves disease, in contrast to Hashimoto’s
thyroiditis, is characterized by hyperthyroidism, a
state of excessive thyroid function. Graves disease
is, in fact, one of the most frequently occurring
autoimmune diseases and the most common cause
of hyperthyroidism. Women exhibit greater
susceptibility to
• Graves disease by a margin of about 5 to 1 and
most often present with the disease in the fifth and
sixth decades of life.
• Clinical symptoms include nervousness, insomnia,
depression, weight loss, heat intolerance, sweating,
rapid heartbeat, palpitations, breathlessness,
fatigue, cardiac dysrhythmias, and
restlessness.84,90 Another sign present in
approximately 35% of patients is exophthalmos, in
which hypertrophy of the eye muscles and
increased connective tissue in the orbit cause the
eyeball to bulge out so that the patient has a large-
eyed staring expression.
• The major antibodies involved in the pathogenesis
of Graves disease are the thyroid-stimulating
hormone receptor antibodies (TRAbs).
SYSTEMIC LUPUS ERYTHEMATOSUS
− Butterfly rush a.k.a. RED WOLF – most
commonly seen
TESTS
◼ ANA – Serological test detecting SLE
➔ Uses indirect immunofluorescence
 MYASTHENIA GRAVIS
Eyes
Tissue-specific affecting neve and
Systemic disease
Neuromuscular disorder
Blocks Acetylcholine binding
Myasthenia gravis (MG) is an autoimmune disease that
affects the neuromuscular junction. It is characterized
by weakness and fatigability of skeletal muscles.
MULTIPLE SCLEROSIS
− Affects CNS (brain, spinal cord)
− Signs and symptoms depends on where is the
affected nerve fiber (e.g. eyes – vision; body –
fatigue)
− Treatment: Beta interferons; copacxone;
natulizumam medication; corticosteroids,
cyclophosphamide, intravenous
immunoglobulin
• MS include reduced exposure to sunlight, vitamin D
deficiency, and cigarette smoking
• MS is characterized by the formation of lesions
called plaques in the white matter of the brain and
spinal cord, resulting in the progressive destruction
of the myelin sheath of axons.
LYMPHOMA
− Affects lymphocyte lineage → type of tumor
− Different ANTIGEN for
screening/staging/diagnosis
− Lactate dehydrogenases – STAGING
− B-2 Microglobulin – STAGING
− B1, LD 20 – therapy, assessment of metastasis
Hodgkin’s Lymphoma – a type of lymphoma where
the lymphocytes grow out of control, causing swollen
lymph nodes and growth throughout the body
- First discovered in 1832 by Dr. Thomas
Hodgkin
- Reed-Sternberg (RS) cells - typically large with
a bilobed nucleus and two prominent nucleoli;
gives the cell an “owl’s-eyes” appearance
- Hodgkin’s cells’ nuclei are not bilobed and have
a single nucleolus.
- Both cells are from B-cell lineage
Non-Hodgkin’s Lymphoma – a type of lymphoma
where the lymphocytes grow abnormally and can form
growths throughout the body
- Both are kinds of tumors
LEUKEMIA
Immunoproliferative disease
Affects WBC lineage
Can also affect RBC (myelogenous) & thrombocyte
lineage
Leukemia are CANCER CELLS
• Myelogenous and Lymphocytic - two groups based
on the cell type from which they originated
• Myelogenous leukemias are derived from the
common myeloid precursor and encompass the
granulocytic, monocytic, megakaryocytic, and
erythrocytic leukemias
• Acute lymphocytic leukemia (ALL) (also known as
acute lymphoblastic leukemia) is characterized by
the presence of very poorly differentiated precursor
cells (blast cells) in the bone marrow and peripheral
blood.
- ALL is usually seen in children between 2 and
5 years of age
• The chronic lymphocytic leukemias or lymphomas
are a group of diseases almost exclusively of B-cell
origin
- They include chronic lymphocytic leukemia
(CLL) and small lymphocytic lymphoma (SLL).
The WHO considers CLL and SLL a single
disease with different clinical presentations.
Both reveal the B-cell marker CD19 but weakly
express CD20.
BRUTON AGGAMMAGLOBULINEMIA
X-linked disease
Deficiency B-cell lineage
Affects males predominantly
PX: esp. X-link Gammaglobulinemia
Lack circulating B-cells → lack Immunoglobulin all
classes
X-linked hypogammaglobulinemia results from arrested
differentiation at the pre–B-cell stage, leading to a
complete absence of B cells and plasma cells.
MULTIPLE MYELOMA
− Plasma cell
− Affecting specific B-cell
• A.k.a. plasma cell myeloma
• Most serious and common of the plasma cell
dyscrasias (characterized by the overproduction of
a single immunoglobulin component called a
myeloma protein (M protein), or paraprotein, by a
clone of identical plasma cells)
• The clinical manifestations of multiple myeloma
are primarily skeletal, hematologic, and
immunologic
DIGEORGE SYNDROME
22q11.2 deletion syndrome small part of chromosome
22 is missing
Cleft Palate
Deficiency of cellular immunity
Quantitative defect of thymocytes where there is not
enough mature T-cells
DiGeorge anomaly is a developmental abnormality of
the third and fourth pharyngeal pouches that affects
thymus development in the embryo. All organs derived
from these embryonic structures can be affected.
Associated abnormalities include mental retardation,
absence of ossification of the hyoid bone, cardiac
anomalies, abnormal facial development, and thymic
hypoplasia. The severity and extent of the in the CGD phenotype by making the neutrophil
developmental defect can be quite variable. incapable of generating an oxidative burst.
The q11 region of chromosome 22 deletion is also CGD was historically diagnosed by measuring the
associated with velocardiofacial syndrome (VFS) and ability of a patient’s neutrophils to reduce the dye
other syndromes. nitroblue tetrazolium (NBT). NBT reduction is caused
by the production of hydrogen peroxide and other
WISKOTT-ALDRICH SYNDROME reactive forms of oxygen.
− Combination of both cellular and hormonal
immunity
− TRIAD of immunodeficiency (IgG), Eczema,
thrombocytopenia (low platelet)
• Wiskott-Aldrich syndrome (WAS) is a rare X-linked
recessive syndrome that is defined by the triad of
immunodeficiency, eczema, and thrombocytopenia.
WAS is usually lethal in childhood because of
infection, hemorrhage, or malignancy. Milder
variants have also been described such as an X-
linked form of thrombocytopenia.

ATAXIA-TELAGIECTESIA
Combination of cell and hormonal immunity
Cerebral atasia
Affects platelets, epithelial tissue
Low or absent IgG (2), IgA, IgE
Ataxia-telangiectasia (AT) is a rare autosomal recessive
syndrome characterized by cerebellar ataxia
(involuntary muscle movements) and telangiectasias
(capillary swelling resulting in red blotches on the skin),
especially on the earlobes and conjunctiva. Blood
vessels in the sclera of the eyes may be dilated and
there may also be a reddish butterfly area on the face
and ears. Ninety-five percent of patients exhibit
increased levels of serum alpha-fetoprotein.
The AT gene is located on chromosome 11, region q22.
This abnormality results in a defective kinase involved
in DNA repair and in cell cycle control.

CHRONIC GRANULOMATOUS DISEASE

Specific disease most common that affects neutrophil
function
Not “kaya” ni neutrophil to generate oxidative burst
Common test: NITRO BLUE TETRAZONIUM TEST
Chronic granulomatous disease (CGD) is a group of
disorders involving inheritance of either an X-linked or
autosomal recessive gene that affects neutrophil
microbiocidal function. The X-linked disease accounts
for 70% of the cases and tends to be more severe.
Symptoms of CGD include recurrent suppurative
infections, pneumonia, osteomyelitis, draining
adenopathy, liver abscesses, dermatitis, and
hypergammaglobulinemia. Typically, catalase-positive
organisms such as Staphylococcus aureus,
Burkholderia cepacia, and Chromobacterium violaceum
are involved. Infections usually begin before 1 year of
age and the syndrome is often fatal in childhood.
Inability of the patient’s neutrophils to produce the
reactive forms of oxygen necessary for normal bacterial
killing.
Oxidative Burst - genetic defect in any of the several
components of the NADPH oxidase system can result
 TRANSPLANTATION IMMUNOLOGY & TUMOR
IMMUNOLOGY (287)
Transplantation immunology – sequence of events
that occurs after an allograft or xenograft is removed
from donor and then transplanted into a recipient.
A major limitation to the success of transplantation is
the immune response of the recipient to the donor
tissue.
Transplantation
− Transfer of cells, tissues, or organs
− 1st human kidney transplant
➔ 1935
➔ Patient died to mistake in blood typing
Types of Transplant
• Autograft – self to self
- transfer of tissue from one area of the body to
another of the same individual
• Isograft – identical twins
- Aka syngeneic graft is the transfer of cells or
tissues between individuals of the same
species who are genetically identical, for
example, identical twins
• Allograft – same species
- transfer of cells or tissue between two
genetically disparate individuals of the same
species
• Xenograft – different species
- transfer of tissue between two individuals of
different species
Immunology of Transplant Rejection
Components of the Immune system involved in graft
rejection:
1. Antigen presenting cells
• Dendritic cells
• Macrophages
• Activated B cells
2. B cells and antibodies
• Preformed antibodies
• Natural antibodies
• Preformed antibodies from prior sensitization
• Induced antibodies
3. T cells
4. Other cells
• Natural killer cells
• Monocytes/Macrophages
The Allograft rejection
The First set response.
When skin from rabbit is applied to another genetically
unrelated animal –
Initially the graft is accepted. Vascularization start.
Remains healthy for 2-3 days.
By 4th day, inflammation starts, Neutrophils,
Macrophages, and Lymphocytes invade. BVs occluded
by thrombi, vascularity diminishes, ischemic necrosis
sets in. graft changes to scab → sloughed off by 10th
day.
This is called ‘first set response’.
The allograft rejection –
The Second set response.
If, in an animal, which has rejected a graft by first set
response, another graft from the same donor is applied
–
The graft is rejected in an accelerated manner.
Necrosis sets in early, graft is sloughed off by 6th day.
This accelerated allograft rejection is called ‘second
set response’.
Mechanism of Allograft rejection
Basis immunological
• Clear from specificity of second set response.
• Accelerated rejection seen only if the second
graft is from the same donor as the first.
• Application of skin graft from another donor
evokes only the first set response.
Allograft accepted if the recipient animal is made
immunologically tolerant.
• If splenic cells of the donor are injected into
recipient fetal or neonatal animal, they will
accept the graft at a later time. This is due to
specific immunological tolerance.
Role of CD4+ and CD8+
• CD4+ differentiate into cytokine producing
effector cells
• Damage graft by reactions similar to DTH
• CD8+ cells activated by direct pathway kill
nucleated cells in the graft
• CD8+ cells activated by the indirect pathway
are self MHC-restricted, thus cannot kill cells in
the graft.
Effector Mechanisms of Allograft Rejection
• Hyperacute Rejection
• Acute Rejection
• Chronic Rejection
Clinical Manifestations of Graft Rejections Chronic Rejection
• Hyperacute 1. Macrophage and T cell mediated
- Within hours 2. Concentric medial hyperplasia
• Acute 3. Chronic DTH reaction
- Within weeks
• Chronic
- Months to years Prevention and Treatment of Allograft Rejection
• Graft recipients immune system may be
HYPERACUTE REJECTION suppressed
− Characterized by rapid thrombotic occlusion of 1. T cells may be inhibited
the graft ✓ Immunosuppressants such as
− Begins within minutes or hours after corticosteroids, azathioprine and
anastomosis cyclosporine
− Pre-existing antibodies in the host circulation ✓ Corticosteroids can lyse immature T cells
bind to donor endothelial antigens in the thymic cortex
− Activates Complement Cascade ✓ Corticosteroids can also block synthesis of
− Xenograft Response cytokines

Clinical Manifestations of Graft Rejection CYCLOSPORINE

HYPERACUTE
• Pre-existing recipient antibodies
- Natural antibodies (IgM)
- Antibodies from prior transfusion, pregnancy or
transplantation (IgG)
• Graft never become vascularized
Hyperacute Rejection
1. Preformed Ab
2. Complement activation
3. Neutrophil margination
4. Inflammation
5. Thrombosis formation
ACUTE REJECTION
− Vascular and parenchymal injury mediated by
T cells and IgG antibodies that usually begin
after the first week of transplantation if there is
no immunosuppressant therapy
− Incidence is high (30%) for the first 90 days
− Acute Vascular and Acute Cellular
Acute Rejection
1. T-cells, macrophage and Ab mediated
2. Myocyte and endothelial damage
3. Inflammation
CHRONIC REJECTION
− Occurs in most solid organ transplants
➔ Heart, kidney, lung, liver
− Characterized by fibrosis and vascular
abnormalities with loss of graft function over a
prolonged period.
− Fibrosis may represent wound healing
following cellular necrosis of acute rejection.
− However, chronic rejection may develop
without evidence that acute rejection have
occurred
Mechanism of Action
• Acts by blocking activation of T cells by inhibiting
interleukin-2 production (IL-2).
• Decreases proliferation and differentiation of T
cells.
TACROLIMUS (FK506)
• Chemically not related to cyclosporine
• Both drugs have similar mechanism of action
• The internal receptor for tacrolimus is
immunophilin (FK-binding protein, FK-BP)
• Tacrolimus-FKBP complex inhibits calcineurin.
SIROLIMUS (RAPAMYCIN)
• It binds to FKBP and the formed complex binds to
mTOR (mammalian Target Of Rapamycin).
• mTOR is serine-threonine kinase essential for cell
cycle progression, DNA repairs, protein translation
• SRL blocks the progression of activated T cells
from G1 to S phase of cell cycle (Antiproliferative
action).
• It does not block the IL-2 production but blocks T
cell response to cytokines.
MUROMONAB-CD3
• Is a monoclonal antibody prepared by hybridoma
technology
• Directed against glycoprotein CD3 antigen of
human T cells
Prevention and Treatment of Allograft Rejection
• Graft recipients immune system may be
suppressed
2. B cells may also be inhibited
3. Antibodies may be removed
4. Induction of tolerance
 Bone Marrow Transplantation
• Used for leukemia, anemia and immunodeficiency,
especially severe combined immunodeficiency
(SCID).
• About 109 cells per kilogram of host body weight,
is injected intravenously into the recipients.
• Recipient of a bone marrow transplant is
immunologically suppressed before grafting.
• Eg. Leukemia patients are often treated with
cyclophosphamide and total body irradiation to kill
all cancerous cells.
• Because the donor bone marrow contains
immunocompetent cells, the graft may reject the
host, causing graft versus host disease (GVHD).

GRAFT VS. HOST DISEASE

• Caused by the reaction of grafted mature T-cells in
the marrow inoculum with alloantigen of the host

• Acute GVHD
- Characterized by epithelial cell death in the
skin, GI tract, and liver

• Chronic GVHD
- Characterized by atrophy and fibrosis of one or
more of these same target organs as well as
the lungs.

XENOGENEIC TRANSPLANTATION
- A major barrier to xenogeneic transplantation
is the presence of natural antibodies that
cause hyperacute rejection.

Tumor Immunology
Tumor
A proliferation of cells that produces a mass rather
than a reaction or inflammatory condition.
Tumors are neoplasm and are described as benign or
malignant.

Tumor vs. Cancer

TUMOR CANCER
Refers to a mass Particularly threatening
A collection of fluid type of tumor
would meet the definition Derived from the Latin
of a tumor word for crab because
Used term for a cancers are often very
neoplasm irregularly shaped
Types: benign and Refers to a new growth
malignant