IMMUNOLOGY DOCKIE NOTES

HISTORY
Table 1.Significant Milestones in Immunology
SCIENTIST DISCOVERY
Louis Pasteur Therapeutic Vaccination: Live,
attenuated chicken cholera, anthrax and
rabbies vaccines; Father of Immunology
Pasteurization; Father of Microbiology
Edward Jenner Smallpox Vaccination;
Demonstrated cross-immunity
Elie Metchnikoff Phagocytosis
Jules Bordet Complement
Robert Kaus Precipitins
Emil Adolf Von Behring Worked on Serum Therapy
Paul Ehrlich Antibody formation theory;
Chemotherapy
Robert Koch Delayed-type hypersensitivity
Susumo Tonegawa Antibody Diversity
(Genetic Principles underlying
generation of Antibodies with different -Snippet from Turgeon
specificities)
Georges Kohler, Cesar First Monoclonal Antibodies
Milstein SIDENOTES
Rosalyn Yalow Radioimmunoaasay
VACCINATION
Frank Macfarlane Burnet, Clonal Selection Theory
➢ Edward Jenner: Smallpox Vaccine
Niels Jerne
➢ Louis Pasteur: Attenuated Vaccines (Cholera, Anthrax, Rabies)
Frank Macfarlane Burnet, Immunologic Tolerance
➢ Attenuation methods: Chemical, Temperature (cold and hot),
Peter Medawar
Aging, In vitro cell passage
Paul Portier, Charles Robert Anaphylaxis ➢ Variolation: First written records of immunological experimentation
Richet Chinese practice of inhaling powder made from smallpox scabs in
Gerald Edelman, Rodney Basic Structure of Immunoglobulins order to produce protection
Porter ➢ The English physician Edward Jenner brought fame to this
Francoise Barre-Sinoussi, HIV procedure when, in 1796, he injected fluid from the cowpox lesions
Luc Motagnier of milkmaid Sarah Nelmes into an 8-year-old boy named James
George Snell, Jean Dausset, Major Histocompatibility Complex Phipps. When Jenner subsequently inoculated the boy with
Baruj Benaceraf smallpox, James did not develop the disease, showing that the
method was a success. Jenner called this procedure “vaccination”
after the Latin word vacca, which means “cow.”
➢ Louis Pasteur was the first person to use the word vaccination in
reference to all immunization procedures (Stevens 4th Ed. p. 456)
DISCOVERY OF HIV
➢ 1983: France (Francois Barre Sinnoussi, Luc Montagnier
➢ 1984: USA (Robert Gallo, Jay Levy)
OTHER SIDENOTES
➢ Mary Mallon was so called “Typhoid Mary” because she was the
cook whom people believed to be a carrier of typhoid, spreading
typhoid fever to over 51 people.
➢ Syphilis was endemic in Haiti (New World) and was subsequently
contracted and carried back to Europe by the crew of Christopher
Columbus.
➢ There are two types of polio vaccines:
✓ Salk Vaccine is a Formalin-killed poliovirus; injected,
stimulates antibody production in serum but not in
mucosa
✓ Sabin Vaccine is a Live-attenuated poliovirus; orally
administered; stimulates production of both IgA
and IgG; 2 week deferral in donors vaccinated
with such

INNATE/NATURAL/NON-SPECIFIC IMMUNITY

Table 2.Lines of Defense

FIRST LINE OF DEFENSE SECOND LINE OF DEFENSE
PHYSICAL CELLULAR COMPONENTS
-Skin, Nasal Hair, Urination, Vaginal -APCs, WBCs (Phagocytes),
Acidity, Cilia of the Respiratory Tract Acute Phase Reactants, Mast
Cells, NK cells
CHEMICAL
-Sweat, Lysozymes, Acids, Tears, HUMORAL COMPONENTS
Saliva -Cytokines, Inflammation,
NORMAL FLORA Complement System
-Competes with pathogens for
nutrient
PHYSIOLOGIC PROCESSES
-Sneezing, Coughing, Vomiting, Gag
reflex, Perspiration, Shivering,
Crying, Urination, Defecation,
Clotting, Yawning, Burping

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IMMUNOLOGY DOCKIE NOTES
MISCELLANEOUS
-Body temperature, Oxygen tension,
Hormonal balance, Age

-Snippet from Robins

-Snippet from Stevens

SIDENOTE:
➢ In 1903, an English physician named Almroth Wright linked the
two theories (Cellular and Humoral) by showing that the immune
response involved both cellular and humoral 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. These serum factors
include specific proteins known as antibodies, as well as other
factors called acute phase reactants that increase
nonspecifically in any infection.

CELLULAR COMPONENTS -Snippet from Stevens

ANTIGEN PRESENTING CELLS 1. Adhesion (Attachment)

-APCs are cellular factors of immunity that process and “present” antigens to a. Direct Interaction
the T-cell receptor -Via Pattern Recognition Receptors/Primitive Pattern
Recognition Receptors
1. Dendritic Cells -Phagocytes use their PRRs/PPRR to recognize and adhere to
-Their main function is to phagocytose antigen and present it to T-helper Pathogen Associated Molecular Pattern (PAMPs are lipid and
cells carbohydrate sequences) on the microbe surface
-Most potent phagocytic cell in the tissue
-Rich in MHC Class II molecules b. Indirect Interaction
-Classified according to their tissue location -Via Opsonization
a. Langerhans cells -Opsonins attach to foreign substance and prepare it for
-Skin and mucous membranes phagocytosis; they may act by neutralizing the surface charge on
b. Interstitial dendritic cells the foreign particle
-Heart, lungs, liver, kidney, and GI tract
c. Interdigitating dendritic cells c. Ingestion (Engulfment)
-T lymphocyte areas of secondary lymphoid tissue and -Pseudopodia surround the pathogen, then fuse to forma a vacuole
thymus called phagosome
-Oxidative burst occurs (Consumption of Oxygen and formation of
2. Macrophages Toxic Products)
-Arise from monocytes -Phagosomes are converted into phagolysosomes by fusion with
-Have specific names according to their particular tissue location lysosomes or cytoplasmic granules
a. Kupffer cells
-Liver d. Digestion (Killing)
b. Microglial cells -Occurs inside the phagolysosomes
-Brain -Nitric Oxide is produced which is toxic to the pathogen
c. Histiocytes
-Connective tissue a. Oxygen-Dependent Mechanism
-NADPH oxidase forms O2- radical (Superoxide) which is
3. B cells toxic to the pathogen
-Superoxide is converted to a more stable oxidative burst
product, H2O2
PHAGOCYTOSIS -Its effect is potentiated by the formation of Hypochlorite
-Physical Contact occurs as neutrophils roll along until they encounter the site ions via the action of myeloperoxidase in the presence of
of injury or infection. They adhere to receptors on the endothelial cell wall of Chloride ions
the blood vessels and penetrate through the tissue by means of diapedesis.
This process is aided by chemotaxis, whereby cells are attracted to the site
of inflammation by chemical substances such as soluble bacterial factors,
complement components or CRP.

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IMMUNOLOGY DOCKIE NOTES
ACUTE PHASE REACTANTS
-Soluble factors that enhance phagocytosis
-Normal serum constituents that increase rapidly by at least 25% due to
infection, injury, or trauma to the tissues
-Produced primarily by hepatocytes (liver parenchymal cells) within 12-24
hours in response to an increase in certain intracellular signaling polypeptides
called cytokines

b. Oxygen-Independent Mechanism
1. C-Reactive Protein
-Involves Defensins which are antibiotic-like peptides made
-Originally thought to be an antibody to the c-polysaccharide of
by phagocytes, especially PMNs
pneumococci
-Involves Digestive enzymes present in the phagolysosomes
-Consists of five identical subunits held together by non-covalent bonds
(e.g. Lysozyme, Lactoferrin)
-Binding with foreign particles is calcium-dependent and non-specific,
and the main substrate is phosphocholine, a common constituent of
SIDENOTES
microbial enzymes
TOLL LIKE RECEPTORS -Can be thought of as a primitive, nonspecific form of antibody molecule
➢ Discovered by Charles Janeway that is able to act as a defense against microorganisms or foreign cells
➢ Toll is a protein originally discovered in the fruit fly Drosophila, until specific antibodies can be produced
where it plays an important role in antifungal immunity in the adult -Most widely used indicator of acute inflammation
fly -A concentration of more than 2.5mg/L has been defined as the threshold
➢ TLR2: Gram + bacteria for high cardiovascular risk. High sensitivity CRP testing has a lower
➢ TLR4: Gram – bacteria detection of 0.01 mg/dL, allowing for measurement of much smaller
➢ The function of TLR10 is not yet known. increases than the traditional latex agglutination screening test
➢ Found in the cell surface: TLR1, TLR2, TLR4, TLR5, and TLR6
➢ Found in the cytoplasm: TLR3, TLR7, TLR8, and TLR9 2. Serum Amyloid A
➢ The highest concentration of these TLRs occurs on monocytes, -Associated with HDL cholesterol, and it is thought to play a role in
macrophages, and neutrophils (Stevens) metabolism of cholesterol; by removing cholesterol-filled macrophages
at the site of tissue injury, serum Amyloid A contributes to the cleaning
up of the area.

3. Complement
-Series of serum proteins that are normally present and whose overall
function is mediations of inflammation

4. Mannose-Binding Protein
-A trimer that acts as opsonin, which is calcium-dependent
-Recognizes foreign carbohydrates from pathogenic microorganisms
-Homologous to C1q

5. Alpha1-Antitrypsin
-Major component of the alpha-band when serum is electrophoresed
-General inhibitor of proteases released from leukocytes, especially
elastase; deficiency of such results to emphysema

6. Haptoglobin
-An alpha-2 globulin that primarily binds irreversibly to free hemoglobin
released by intravascular hemolysis

7. Fibrinogen
-Most abundant coagulation factor; cleaved by thrombin to form fibrin clot

8. Ceruloplasmin
-Principal copper-transporting protein in human plasma
-Acts as ferroxidase, oxidizing iron from Fe2+ to Fe3+; this may serves as
a means of releasing iron from ferritin for binding to transferrin
-Deficiency is associated with Wilson’s Disease

-Snippet from Stevens NATURAL KILLER CELLS

-Also known as Large Granular Lymphocytes, Null cells, “Kiss of death”
-Naturally occurring, not induced by immunologic insult
-Forms rosettes in sheep RBC
-Express receptors for the Fc portion of IgG; CD16 is the Fc receptor for IgG
-They recognize and lyse antibody-coated cells through a process called
Antibody-Dependent Cell Cytotoxicity whereas binding occurs through the
CD16 receptor for IgG
-CD16 is the Fc receptor for IgG, hence, NK cells are able to lyse selectively
those cells that are coated with antibodies (Antibody-Cell-Mediated-
Cytotoxicity [ADCC]), which is important in some hypersensitivity reactions
-Killing Methods: Perforins (Formation of pores), Activated apoptosis
-Snippet from Stevens (Cytolysis), Granzymes

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IMMUNOLOGY DOCKIE NOTES
-Characterized by the surface markers, CD16 and CD56
-Lymphokine Activated Cell (LAK) is an NK cell exposed to IL-2 and IFN ✓ IFN-beta (Fibroblast/Fibroepithelial Interferon)
gamma -Major producer: Fibroblast and Epithelial cell
-Percentage in the circulation: 5-10% (Stevens); 10-15% (Henry)
b. Type II IFN
✓ IFN-gamma (Immune Interferon)
-Produced by immulogically stimulated Lymphocyte
-Major Producer: T cells
-Stimulates Antigen Presentation by MHC I and MHC II
molecules

3. Tumor Necrosis Factor

-Principal mediator of the acute inflammatory response to LPS in Gram
Negatice bacteria
✓ TNF-alpha (Cachectin)
-Produced by Macrophages and NK cells
-Action: Local inflammation

✓ TNF-beta (Lymphotoxin)
-Produced by T cells (CD4 and CD8) and B cells
HUMORAL COMPONENTS -Action: Killing of target cells

CYTOKINES INFLAMMATION
-Small soluble proteins that regulate the immune system, orchestrating both -The overall reaction of the body to injury by an infectious agent
innate immunity and the adaptive response to infection
-Produced by cells of both the acquired immune system (lymphocytes) as well 1. Rubor
as the cells of the innate system (macrophages, mast cells, etc.). Other cells -Redness due to dilation of blood vessels
(e.g., fibroblasts) not considered classically belong to the immune system per 2. Calor
se, can also produce such molecules. -Heat due to increased blood content
3. Tumor
1. Interleukins -Swelling due to increased capillary permeability
a. IL-1 4. Dolor
-Mediator of host inflammatory response to infections and other -Pain due to increased pressure
inflammatory stimuli 5. Functio Laesa
-Acts as an endogenous pyrogen and induces fever in the acute -Loss of function
phase response through its actions on the hypothalamus

b. IL-2
-Formerly “T-cell Growth factor”
-Has high capacity to induce activation of almost all clones of
cytotoxic cells
-For proliferation of T cells and B cells
-Stimulates Lymphokine-Activated Cell (LAK)

c. IL-3
-Formerly “Multicolony Colony-Stimulating Factor
-Promotes differentiation of hematopoietic cells into all known
mature cell types (AKA Pan-specific Hematopoietin)

d. IL-4
-B cell growth factor I
-Governs B-cell isotype switching to IgG1 and IgE
-Key regulator in humoral and adaptive immunity

e. IL-5
-B Cell Growth Factor II
-Activates Eosinophils and serves as link between T cell activation
and eosinophilic inflammation
-Stimulates growth and differentiation of eosinophils and activates
mature eosinophils (IL-5 is expressed on eosinophils)

f. IL-8
-Potent stimulator of Neutrophils in chemotaxis
-Activates respiratory burst

g. IL-10
-Potent suppressor of macrophage functions
-Antagonist to IFN-gamma, it is a down regulator of the immune
response

2. Interferons
-Discovered in virally infected cultured cells; This interference with viral
replication in the cells by another virus led to the name “interferons”
-Antiviral; produced by dendritic cells

a. Type I IFN
✓ IFN-alpha (Leukocyte Interferon)
-Produced by virus-induced Leukocyte culture -Snippet from Robins
-Major producer: NK cells/ Null Cells

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IMMUNOLOGY DOCKIE NOTES
CLASSICAL PATHWAY

-Snippet from Robins

ALTERNATIVE PATHWAY

Figure X. PGs and their role in inflammation (Robins)

COMPLEMENT SYSTEM
-Series of serum proteins that are normally present and whose overall function
is mediation of inflammation
-Measurement of complement components has two major uses in clinical
diagnosis:
1. To detect congenital deficiencies
2. To detect Acquired complement deficiencies THE THREE PATHWAYS

Table 5. Proteins of the Complement System

CLASSICAL PATHWAY
C1q Binds to Fc of IgM and IgG
C1r Activates C1s
C1s Cleaves C4 and C2
C4 Part of C3 convertase
C2 Binds to C4b- forms C3 convertase
C3 Key intermediate in all pathways
C5 Initiates membrane attack complex
C6 Binds to C5b complex
C7 Binds to C5bC6 in MAC
C8 Starts pore formation on membrane
C9 Polymerizes to cause cell lysis
ALTERNATIVE PATHWAY
Factor B Binds C3b to form C3 convertase
Factor D Cleaves factor B
Properdin Stabilizes C3 convertase
C3 through C9 Table 6.Regulatory Proteins of the Complement System
MBL PATHWAY REGULATION OF THE CLASSICAL PATHWAY
MBL (Similar to C1q) Binds to mannose C1 Inhibitor (C1INH) Dissociates the subunits C1q, C1r and
MASP-1 (Similar to Cr) Unknown (Helps cleave C4 and C2) C1s
MASP-2 (Similar to C1s) Cleaves C4 and C2 Deficiency of C1INH results in HANE
C3 through C9 REGULATION OF C3 CONVERTASE
***This table was adapted and modified from Stevens’ and Navarro’s table Factor I Inactivates C4b and C3b
Involved in both classical and alternative
pathway
Factor H Cofactor with factor I to inactivate C3b
Prevents binding of B to C3b
Involved in alternative pathway
C4 binding protein Acts as a cofactor with I to inactivate C4b
Involved in classical pathway
DAF (CD55) Prevents assembly of both C3
convertase by promoting dissociation of
C2a from C4b and Bb from C3b

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IMMUNOLOGY DOCKIE NOTES
DAF is deficient or absent in cells from -Because all proteins from C1 to C9 are necessary for this to
people with PNH, the RBCs of these occur, absence of any one component will result in an
patients have increased sensitivity to abnormal CH50, essentially reducing this number to zero
complement mediated lysis -Lytic activity can also be measured by radial hemolysis in
Membrane Cofactor Protein Serves as a cofactor of I to inactivate C4b agarose plates. Rabbit red blood cells that have been
(CD46) and C3b sensitized with antibody are implanted in agarose, and patient
CR1/Complement Receptor Acts as a receptor on platelets and RBCs serum is added to wells punched in the gel. Lysis appears as
Type 1 and to mediate transport of C3b-coated a clear zone around each well, and if complement standards
(CD35) immune complexes to the liver and are run, the size of the zone can be related to complement
spleen concentration.
REGULATION OF THE MAC
MIRL (CD59) Binds to C8 and prevents the interaction
***Other reference call this: of C8 with C5b67
Protectin/HRF
S protein (Vitronectin) Prevents attachment of the C5b67
complex to cell membranes

Table 7.Deficiencies of Complement Components

DEFICIENT ASSOCIATED DISEASE
COMPONENT
C1 (q,r,s) Lupuslike Syndrome; Recurrent infections
C2 Lupuslike Syndrome; Recurrent infections;
Atherosclerosis
C3 Severe recurrent infections;
Glomerulonephritis
(Henry: Glomerulonephritis: deficiency in
C1qrs, C2, C4 and Factor H)
C4 Lupuslike Syndrome
C5-C8 Neisseria infections
C9 No known disease association (Stevens and
Henry)
C1INH Hereditary Angioedeme (HANE)
DAF (CD 55) Paroxysmal Nocturnal Hemoglobinuria
MIRL (CD 59) Paroxysmal Nocturnal Hemoglobinuria
Factor H or Factor I Recurrent Pyogenic Infections
MBL Pneumococcal diseases, sepsis, Neisseria
infections
Properdin Neiseria infections
MASP-2 Pneumococcal diseases

LABORATORY DETERMINATION OF COMPLEMENT ABNORMALITIES

1. Immunologic Assays for Individual Components

-Components that are usually measured and for which there are
standardized reagents include C1q, C4, C3, C5, factor B, factor H,
factor I, and C1 inhibitor. Kits are available for C3a, C4a, and C5a.
-None of the assays for individual components are able to
distinguish whether the molecules are functionally active.
a. RID
-uses agarose gel into which specific antibody is incorporated
-Serum serves as the antigen and is placed in wells that are
cut in the gel.
-Diffusion of the antigen from the well occurs in a circular
pattern.
-The radius of the resulting circle can be related to antigen
concentration.
-This is a sensitive technique when performed correctly, but
it takes at least 24 hours before test results are available.
b. Nephelometry
-Measures concentration according to the amount of light
scattered by a solution containing a reagent antibody and a
measured patient sample
-Generally, the more antigen–antibody complexes that are
present, the more a beam of light will scatter as it passes
through the solution
-Such systems have a high degree of accuracy, results are
available quickly, and processing is easy because of the use
of automation. However, it is necessary to purchase
expensive equipment.
2. Assays for the Classical Pathway
a. CH50 (Hemolytic Titration Assay)
-Measures the amount of patient serum required to lyse 50
percent of a standardized concentration of antibody-
sensitized sheep erythrocytes.
6 MARCJAYGAGARIN, RMT, MD :3 “A human’s belief can make the impossible, possible. “ –Akihisa Yoshi
b. ELISA
-Solid-phase IgM attached to the walls of microtiter plates is
used to initiate complement activation. Antihuman antibody to
C9 conjugated to alkaline phosphatase is the indicator of
complement activation. When a substrate is added, if any C9
is present and the antibody conjugate has attached, a color
change will be evident.
3. Assays for the Alternative Pathway
a. AH50
-Performed in the same manner as the CH50, except
magnesium chloride and ethylene glycol tetraacetic acid are
added to the buffer, and calcium is left out; This buffer
chelates calcium, which blocks classical pathway activation;
Rabbit red cells are used as the indicator, because these
provide an ideal surface for alternative pathway activation
b. ELISA
-Microtiter wells are typically coated with bacterial
polysaccharide to trigger activation of the alternative pathway
4. Assay for the Three Pathways
a. Reagent Strip
-Strips used for the classical pathway are coated with IgM,
strips for the alternative pathway are coated with
lipopolysaccharide, and strips for the mannose-binding
lectin pathway are coated with mannose.
-Such testing is easy to perform and is not dependent upon
the use of animal erythrocytes, which may be hard to obtain.
IMMUNOLOGY DOCKIE NOTES
-Deficiencies can be detected using the combined test Table 11.Examples of Active and Passive Immunity
results. ACTIVE IMMUNITY PASSIVE IMMUNITY
NATURAL
5. Complement Fixation Test
-Detects complement consumption in any cellular or non-cellular ARTIFICIAL
antigen-antibody reaction to which complement is bound
-Used in the detection of viral, fungal, and rickettsial antibody
-When antigen-antibody occurs, complement is ‘inactivated’ or
fixed such that it will be unavailable to cause lysis upon addition of
indicator cells (SRBCs coated with amboceptor/hemolysin) PRIMARY LYMPHOID ORGANS
1. BONE MARROW
-B cells received their name because they were originally found to
mature in birds in an organ called the bursa of Fabricius, which is
similar to the appendix in humans. After searching for such an
organ in humans, it was discovered that B-cell maturation takes
place within the bone marrow itself. Thus, the naming of these cells
was appropriate (Stevens 4th Ed.)

2. THYMUS
-T cells develop their identifying characteristics in the thymus,
which is a small, flat, bilobed organ found in the thorax, or chest
cavity, right below the thyroid gland and overlying the heart. In
humans, the thymus reaches a weight of 30 to 40 g by puberty and
then gradually shrinks in size
-Although the thymus diminishes in size as humans age, it is still
capable of producing T lymphocytes, although at a diminished rate
-Maturation of T cells takes place over a 3-week period (Stevens
4th Ed.)

SECONDARY LYMPHOID ORGANS

1. SPLEEN

SIDENOTES: 2. LYMPH NODES

➢ The RICE test or indirect complement fixation test serves to
detect non-complement-binding antibodies (antibodies that
do not bind with complement)
➢ The non-lytic complement fixation test or conglutinating
adsorption test uses conglutinin which is a beta-globulin derived
from cattle serum. The Ag-Ab complexes are aggregated by
conglutinin

ADAPTIVE/ACQUIRED/SPECIFIC IMMUNITY
-Self/Non-self-Discrimination
-Memory, Specific
-Immunity can be Active or Passive

Table 9. Adaptive Immunity

THIRD LINE OF DEFENSE
CELLULAR COMPONENTS: HUMORAL COMPONENTS:
-T cels, B cells*, Plasma cells* -Antibodies, Cytokines
*Classified under humoral components by other medical literatures

Table 10. Two Types of Adaptive/Acquired Immunity

ACTIVE IMMUNITY PASSIVE
IMMUNITY
Source 3. MALT

Immunizing agent
Relative length of
immunity
Effectiveness
Undesirable effect

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IMMUNOLOGY DOCKIE NOTES
-Rearrangement of genes coding for Heavy chain at
chromosome 14
-Expression of CD19 and CD45R

b. Pre-B cell (Precursor B cell)

-Rearrangement of genes coding for Light chains at
chromosomes 2 and 22
-Expression of mu chain in the cytoplasm

c. Immature B cell
-Expression of IgM on the surface, CD21, CD35 and CD40
-Expression of MHC class II

d. Mature B cell (Marginal Zone B cells)

-Expression of IgD in the surface in addition to IgM
-Resting (naïve)

e. Activation of B cell
-Occurs in the secondary lymphoid organ (e.g. In the
Germinal center of the secondary follicle of lymph node)
-Contains CD25 which is a receptor of IL-2

f. Plasma cell
CELLULAR COMPONENTS -Result of antigenic stimulation and transformation of activated B
cells
T CELLS -Secrete Antibodies
1. Development/Maturation
a. In the Cortex: Table 12.T Lymphocytes versus B Lymphocytes
• Double-Negative Lymphocyte Stage T CELLS B CELLS
-Rearrangement of the genes that code for TCR (CD3) Site of Maturation Thymus Bone Marrow
-Rearrangement of the beta chain (Bursa of Fabricius
-Expression of CD2, CD5 and CD7 in bird)
Mature stage of Th, Tc, Ts, Tm Plasma cell
• Double-Positive Lymphocyte Stage Differentiation
-Rearrangement of the alpha chain Location in Lymph Cortex Paracortex
-Expression of CD4 and CD8 Nodes
o Positive Selection Present in the following Blood, Thoracic duct Bone Marrow,
-Only Double-Positive cells with functional organs fluid, Spleen, Lymph Spleen, Lymph
TCR are allowed to survive/remain Nodes Nodes
-Any thymocytes that are unable to Relative Concentration 60-80% 10-20%
recognize self-MHC antigens die without in Peripheral Blood
leaving the thymus Products of Activation Cytokines: Antibodies
Lymphokines
o Negative Selection Portion of conjugated Carrier Hapten
-Takes place among the “surviving” double- Ag to which reactivity is
positive T cells primarily detected
-Most T cells that would be capable of an Relative Life Span Long (4-6 years) Short (3-4 days)
autoimmune response are eliminated in this General Function Cell-mediated Humoral-mediated
manner immune response immune response
-Only 1-3% of the double-positive T cells in APC
the cortex survive CD markers/Antigens CD2, CD3, CD4, CD8 CD19, CD20,
-“Survivors of selection exhibit only one type CD2: SRBC receptor, CD21, CD40,
of marker, either CD4 or CD8 and they The classical T cell MHC class II
migrate to medulla surface marker)
CD3: Part of TCR
b. In the Medulla (1984)
-Mature T cells CD4: Receptor for
• CD4+ cells MHC II
-Recognize antigens along with MHC class II CD8: Receptor for
-Termed helper or inducer cells MHC I
-2/3 of the peripheral T cells Antigen Receptor TCR BCR
• CD8+ cells ***This table was adapted and slightly modified from Turqueza’s table
-Recognize antigen along with MHC class I
-Termed cytotoxic or suppressor cells
-1/3 of the peripheral T cells LABORATORY IDENTIFICATION OF LYMPHOCYTES
• T-helper subsets 1. FICOLL-HYPAQUE DENSITY GRADIENT CENTRIFUGATION
o Th1 produces IFN-gamma and TNF-beta; -Whole blood diluted with buffer (Defibrinated blood or Heparinized
Activates CD8+ cells and Delayed type blood) is layered onto Ficoll-Hypaque medium in a plastic
hypersensitivity reaction centrifuge tube. Tubes are spun at 400 x g for 30 minutes. Red
o Th2 produces IL-4 and IL-5; Activates B cells to blood cells and granulocytes settle to the bottom of the tube, while
produce Ab (IgE) mononuclear cells (monocytes and lymphocytes) form a band at
o Treg the interface of the Ficoll-Hypaque and plasma.
-Ficoll-Hypaque is available commercially and has a specific
B LYMPHOCYTES gravity that varies between 1.077 and 1.114, depending on the
1. Development/Maturation manufacturer

a. Pro-B cell (Progenitor B cell)

-Occurs in the bone marrow

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IMMUNOLOGY DOCKIE NOTES
LABORATORY EVALUATION OF LYMPHOCYTE FUNCTION

1. LYMPHOCYTE TRANSFORMATION ASSAYS

A. MITOGEN-INDUCED BLASTOGENESIS
-In vitro lymphocyte transformation in response to a mitogen
was first reported by Nowell (1960).
-Mitogens and IL-2 in general are the most potent stimuli, as
they induce the rapid proliferation of the largest proportion of
lymphocytes. For this reason, proliferation can be detected in
3–4 days and thus provides an effective screening tool. Of the
mitogens, phytohemagglutinin and concanavalin A (Con
A) induce T cell blastogenesis, and pokeweed mitogen
and staphylococcal protein A trigger B cell activation.
-Lymphocytes are isolated with Ficoll-Hypaque density
gradient centrifugation and cultured for several days with a
mitogen or a specific soluble antigen in media enriched with
human AB serum. Tritiated thymidine is added to the culture
media, and the plates are incubated for an additional 4–24
A. AUTOMATED METHOD hours. The cells are isolated (harvested) from the plates using
• Flow Cytometry a special device (cell harvester) and are transferred to disks
-Once a lymphocyte population has been obtained, of filter paper.
segregation into subsets is accomplished via flow cytometry. -These disks are placed in scintillation fluid, and radioactive
Cell flow cytometry is an automated system for emissions are measured in a liquid scintillation counter. This
identifying cells based on the scattering of light as cells technique is classified as a bulk assay method because the
flow in single file through a laser beam. response of the entire cell population is measured.
***Intrinsic Parameters: FSC, SSC -The results are expressed as a stimulation index, which is
FORWARD-ANGLE LIGHT ORTHOGONAL RIGHT ANGLE the ratio of radioisotope incorporation into the test versus
SCATTER (FSC) LIGHT SCATTER control cells. The relative proliferation index is the ratio of the
[90-degree-angle light scatter Δcounts per minute (cpm) of the test subject (cpm of
(SSC)] stimulated cells minus unstimulated control cells) to the Δcpm
-FSC, or light scattered at less -SSC signal is indicative of of a panel of normal individuals tested simultaneously.
than 90 degrees, is considered granularity or intracellular
an indicator of size complexity of the cell B. MIXED LYMPHOCYTE CULTURE
***Extrinsic Parameter -Mixed lymphocyte culture assay (MLC) is a special type of
Unlike FSC and SSC, which represent light-scattering properties that are lymphocyte stimulation assay based on the ability of
intrinsic to the cell, extrinsic parameters require the addition of a fluorescent histoincompatible lymphocytes from one individual to
probe for their detection. Fluorescent-labeled antibodies bound to the cell can stimulate the lymphocytes of another individual (mixed
be detected by the laser. lymphocyte reaction)
-The D locus of the HLA system is the major determinant of
• IFA the MLC phenomenon. When two cells share common D loci,
-Fluorescent antibodies are used to screen for they are not able to stimulate each other, but when the D loci
subpopulations, such as B cells, T helper cells, and T are different, the cells are stimulated. The MLC is unilateral
cytotoxic cells. The antibodies used are monoclonal, and (one way) when one group of cells are made incapable of
each has a different fluorescent tag. responding (by treatment with radiation or mithromycin); it is
bidirectional (two way) when no radiation or mithromycin is
utilized. In the one-way MLC, untreated cells are termed the
B. MANUAL METHOD responder population, and treated cells are the stimulator
• Rosette Technique population
-Rosette test uses sheep red blood cells that attach to -The principal clinical use of the MLC assay is to assist in the
lymphocytes (Sheep red cells attach to CD2 Ag which is selection of a compatible donor for a bone marrow transplant,
only found in T cells) because the MLC is a predictor of host response to a
-Lymphocytes are separated from the Whole blood and mixed transplanted organ. To prevent graft rejection or graftversus-
with sheep red cells. host disease, donor and recipient cells must be mutually non-
stimulatory

-Snippet from Stevens

9 MARCJAYGAGARIN, RMT, MD :3 “A human’s belief can make the impossible, possible. “ –Akihisa Yoshi
IMMUNOLOGY
Figure X. One-way mixed lymphocyte culture assay (MLC). Irradiated
stimulator cells, which are incapable of cell division, are mixed with non-
irradiated responder cells, which can proliferate and divide. After incubation
under cell culture conditions for approximately 1 week, responder cells
proliferate in the presence of a disparity in the human leukocyte antigen (HLA)-
D locus (right side of diagram), but no proliferation occurs if the responder and
stimulator cells are HLA-D compatible (left side of diagram). Tritiated
thymidine is added to the reaction mixture, and the degree of proliferation is
determined by liquid scintillation counting.(Henry)
2. CYTOTOXICITY ASSAYS
Cell death (cytotoxicity) is the endpoint commonly used in
functional assays of the cellular immune system. In these assays,
cell cytotoxicity may occur as the result of complement activity
(complement-mediated cytotoxicity) or may be due to the direct
effect of one cell on another (cell-mediated cytotoxicity).
Conventionally, target cell lysis is determined by the release of a
substance such as 51chromium (51Cr) from the target cell upon
death, or by the incorporation of a vital dye such as eosin or tryptan
blue. A variation of these assays, the microlymphocytotoxicity
assay, is widely used in the clinical histocompatibility laboratory for
human organ transplantation.
A. CYTOTOXIC T CELL (CTL) ASSAYS
-Cytotoxic T cells (CTLs, T killer cells, cytolytic T cells, CD8+
T cells, killer T cells) constitute a subset of T cells important
in the host defense against altered class I MHC-positive self
cells, especially cells infected with a virus or other pathogen.
CTLs recognize altered target cells by the presence of
peptide fragments produced by processing of the foreign
antigen and presented by the class I MHC receptor to the T
cell antigen receptor. Recognition of an MHC complex on a
target cell by a CTL induces cell death by apoptosis by the
granule-exocytosis pathway (granule endocytosis) or the
FAS-FAS ligand pathway.
-The chromium release assay has become the standard
technique for the measurement of complement- or cell-
mediated cytotoxicity.
10 MARCJAYGAGARIN, RMT, MD :3 “A human’s belief can make the impossible, possible. “ –Akihisa Yoshi
DOCKIE NOTES
Figure X. Principle of the antibody-dependent cell-mediated cytotoxicity
(ADCC) assay. This assay is performed with mononuclear cells from a kidney
organ donor or from several unrelated individuals (panel cells), which are
stimulated with phytohemagglutinin (PHA) and labeled with 51Cr. Following
incubation with eluate from another nephrectomy specimen or from normal
human serum, labeled cells are washed and incubated with effector cells from
a normal unrelated individual. Radioactivity (γ-emissions) in the cells at the
end of the experiment represents
unlysed cells. The results are expressed as % cytotoxicity. (Henry)
A. NATURAL KILLER CELL-MEDIATED CYTOTOXICITY
NK cells are critical members of the innate immune system
characterized by their capability to lyse target cells without
prior sensitization, termed spontaneous cell-mediated
cytotoxicity. NK cells can be subdivided into two functional
and immunophenotypically distinct groups defined by the
surface density of CD56. Release of 51Cr from labeled target
cells is the standard method for the characterization of NK
cells.
B. MICROLYMPHOCYTOTOXICITY ASSAY
-The dye exclusion lymphocytotoxicity assay is the standard
technique for the detection of an antibody–antigen interaction
on a cell surface. The lymphocytotoxicity assay was
introduced by Terasaki and McClelland in 1964.
-Viable cells (usually lymphocytes) are incubated with serum-
containing antisera. If a cell surface antigen is present that is
recognized by antibodies in the sera, an antigen–antibody
complex will form on the surface. These complexes are
detected by the sequential addition of rabbit complement and
a vital dye, such as eosin, to the reaction mixture. The
occurrence of complement fixation on the cell membrane
leads to activation of the terminal complement components,
and eventually to cell lysis and death. Dead cells are detected
and counted by phase microscopy after differential uptake of
the eosin dye and fixation with formalin. Antibody-bound
lymphocytes will die, take up the eosin dye, and give a
positive reaction; unbound lymphocytes will remain viable,
exclude the eosin dye, and give a negative reaction (dye
exclusion).
IMMUNOLOGY DOCKIE NOTES
d. Fragments

3. Treatment with Proteolytic Enzymes

a. Cleavage with Pepsin (Alfred Nisonoff)

b. Cleavage with Papain (Porter)

Figure X. Principle of microlymphocytotoxicity by dye exclusion. Cells

and serum are incubated together in the wells of a microtiter tray. Mineral oil
at the top of the wells prevents fluid evaporation. Complement is added, and
a second incubation is performed. The cells are washed, eosin is added, and
the cells are fixed with formalin. Cells that fixed antibody and were killed
through the action of complement take up eosin and appear dark and
nonrefractive under a phase-contrast microscope. In contrast, living cells are
refractive and are easily identified by phasecontrast microscopy. Variations of
the microlymphocytotoxicity assay are used for human leukocyte antigen
(HLA) typing, HLA crossmatch, and detection of anti-HLA antibodies. (Henry)

HUMORAL COMPONENTS

ANTIBODIES
1. Theories on Antibodies
a. Side Chain Theory
-Certain cells had specific surface receptors for antigen that were
present before contact with antigen occurred; Once antigen was 4. Properties of Immunoglobulins
introduced, it would select the cell with the proper receptors, Table 14.Properties of the Five Immunoglobulins
combination would take place, and then receptors would break off IgG IgM IgA IgD IgE
and enter the circulation as antibody molecules Constant Gamma Mu Alpha Delta Epsilon
Heavy chain
Structure/For Monomer Pentamer Dimer Monomer Monome
b. Template Theory/Instructive Theory m Monomer Monomer r
-Antibody producing cells are capable of synthesizing a Abundance Most Least
generalized type of antibody, and when contact with an antigen abundant Abundan
occurs, the antigen serves as a mold or template and alters protein t
MW Lightest Heaviest
synthesis so that antibody with a specific fit is made and will enter (150 000 Da) (900 000
the circulation as the antigen remains behind to direct further Da)
synthesis Sedimentation 7S 19S 7S 7S 8S
Coefficient
Serum Half- Longest Shortest
c. Clonal Selection Theory life Half-life Half-life
-Individual lymphocytes are genetically preprogrammed to produce Protein J chain J chain
one type of immunoglobulin, and that a specific antigen finds or Attachments
selects those particular cells capable of responding to it, causing it Distribution Intravascular Intravascula Intravascular Membran Fc
Extravascula r , e Epsilon
to proliferate r Mucosa (Naïve B Receptor
cells) on mast
2. General Structure cells,
a. Chain basophil
s
Subclasses 4 2
Breast Milk Present Absent Present Absent Absent
b. Domain (Colostrum) (Majority)
Placental Yes No No No No
Transfer
Stable at Yes Yes Yes No No
c. Disulfide bond 56deg C
Complement Yes Yes No No No
Fixation (Stronger)

11 MARCJAYGAGARIN, RMT, MD :3 “A human’s belief can make the impossible, possible. “ –Akihisa Yoshi
IMMUNOLOGY DOCKIE NOTES
*Can
Activate
Complement
via Alternate
Pathway
Agglutination Moderate Strong Weak ? No
Precipitation Strong Variable Weak ? No
Opsonization Weak Strong ? ? ?

CYTOKINES

12 MARCJAYGAGARIN, RMT, MD :3 “A human’s belief can make the impossible, possible. “ –Akihisa Yoshi
IMMUNOLOGY DOCKIE NOTES
c. Heteroantigens are from other species such as other animals,
MISCELLANEOUS TOPICS plants, or microorganisms

ANTIGENS d. Heterophile antigens are heteroantigens that exist in unrelated

1. TERMINOLOGIES plants or animals but are either identical or closely related in
a. Immunogens structure so that antibody to one will cross-react with antigen of the
-Macromolecules capable of triggering an adaptive immune other; From unrelated plants or animals that can cross react with
response by inducing the formation of antibodies or sensitized T Ag of another
cells in an immunocompetent host
MAJOR HISTOCOMPATIBILITY COMPLEX (HLA)
b. Antigens -Ability of the immune system to recognize/discriminate self from non-self is
-Substance that reacts with antibody or sensitized T cells but may the role of MHC
not be able to evoke an immune response in the first place -Antigen presentation in association with MHC is an absolute requirement for
T cell activation in an immune response
c. Epitope -Genes coding for MHC molecules in humans are found on the short arm of
-Antigenic determinant; Part of an antigen that reacts specifically chromosome 6
with an Antibody or T-lymphocyte receptor
d. Haptens Table 17.MHC Classes
-Non-immunogenic materials that, when combined with a carrier, CLASS I CLASS II CLASS III
create new antigenic determinants Loci

e. Schlepper Molecule
-Carrier Molecule; Couples with haptens which will confer new
antigenic specificities Structure made
up of
f. Allergen Distribution
-Special class of immunogen that induces hypersensitivity
reactions
Function
g. Adjuvant
-Substance administered with an immunogen that increases the
immune response

2. FACTORS AFFECTING IMMUNOGENECITY Table 18.Relationship of Certain HLA and Diseases

Table 16.Factors Affecting Immunogenecity Ankylosing Spondylitis B27
Foreignness Foreignness is the degree to which antigenic Reiter’s Syndrome B27
determinants are recognized as nonself by an Congeinital Adrenal Hyperplasia B47
individual’s immune system. The immunogenicity of Behcet’s Disease B5 (Bw51)
a molecule depends to a great extent on its degree
Psoriasis Vulgaris Cw6
of foreignness.
Type 1 Diabetes DR3
Molecular Weight The higher the MW, the better the molecule will
Rheumatoid Arthritis DR4
function as an antigen. Usually, an immunogen
must have a molecular weight of at least 10,000 Gold-induced Nephropathy DR5
Daltons to be recognized by the immune system. Chronic Lymphatic Leukemia DR5
Chemical Proteins and polysaccharides are the best Kaposi’s Sarcoma (Mediterranean) DR5
Composition immunogens. Carbohydrates are somewhat less ***This table was adapted from Turgeon’s
immunogenic than protein, because the units of
sugars are more limited than the number of amino Table 19. Relationship of Human Leukocyte Antigens to Risk of Disease
ANTIGEN RELATED DISEASES RISK
acids in protein. Pure nucleic acids and lipids are not PRESENT
immunogenic by themselves except when they are B27 Ankylosing Spondylitis 100x
attached to a suitable carrier molecule. Reiter’s Syndrome 40x
Physical form In general, particulate antigens are more Anterior Uveitis 25x
immunogenic than soluble ones. Denatured Arthritic infection with Salmonella or Yersinia 20x
Psoriatic arthritis with spinal involvement 11x
antigens are more immunogenic than the native Spondylitis associated with inflammatory bowel 9x
form. disease 5x
Degradability Antigens that are easily phagocytosed are generally Juvenile chronic arthritis with spinal
more immunogenic. If a macromolecule can’t be involvement
degraded and presented with MHC molecule, then B8 Celiac Disease 9x
Addison’s Disease 6x
it would be a poor immunogen. Myasthenia Gravis 5x
Antigen For a substance to elicit an immune response, it Dermatitis Herpetiformis 4x
Presentation must be subject to antigen processing, which Chronic Active Hepatitis 4x
involves enzymatic digestion to create small Sjogren’s Syndrome 3x
peptides or pieces that can be complexed to MHC DM type I 2x
Thyrotoxicosis 2x
molecules to present to responsive lymphocytes. B5 Behcet’s Syndrome (Disease) 6x
Route, Dosage Generally, intravenous and intraperitoneal routes BW38 Psoriatic Arthritis 7x
and Timing are effective; the intradermal route offers stronger Bw15 DM type I 3x
stimulus than the subcutaneous or intramuscular DR2 Goodpasture’s Syndrome 16x
route. Multiple Sclerosis 4x
Generally, the smaller the dose, the less likely a DR3 Gluten-Sensitive Enteropathy 21x
Dermatitis Herpetiformis 14x
response. Subacute Cutaneous Lupus Erythematosus 12x
Addison’s Disease 11x
3. RELATIONSHIP OF ANTIGENS TO THE HOST Sjogren’s Syndrome (Primary) 10x
a. Autoantigens are those antigens that belong to the host (Self DR4 Pemphigus (In Jewish Persons) 32x
antigens) Giant Cell Arthritis 8x
Rheumatoid Arthritis 6x
DM type I 5x
b. Alloantigens are from other members of the host’s species and DR5 Pauciarticular Juvenile Arthritis 5x
these are capable of eliciting an immune response Scleroderma 5x
Hashimoto’s Thyroiditis 3x
***This Table was adapted from Turgeon’s
13 MARCJAYGAGARIN, RMT, MD :3 “A human’s belief can make the impossible, possible. “ –Akihisa Yoshi
IMMUNOLOGY DOCKIE NOTES
HYPERSENSITIVITY Individual was
-Normal but exaggerated or uncontrolled immune response to an antigen previously
that can produce inflammation, cell destruction or tissue injury exposed to donor
MHC antigens
Accelerated 2-5 days Cell-Mediated Occurs after
Table 20.Types of Hypersensitivity Reactions
transplantation of
TYPE I TYPE II TYPE III TYPE IV
Alternate Anaphylactic/ Cytotoxic/ Immune Delayed/
second graft
name Immediate Antibody- Complex/ Cell-Mediated which shares a
Mediated Complex- significant number
Mediated of antigenic
Immune IgE IgG or IgM IgG or IgM T cells determinants with
Mediator the first one,
Complement No Yes Yes No results in a rapid
Involvement rejection;
Effector Cells Basophils, Red cells, Host Tissue Macrophage and T
Mast cells WBCs, cells cells
Cause is due to
Platelets (Macrophag presence of
e, Mast sensitized T cells
Cells) during the first
Immune Release of Cytolysis due Deposits of Release of graft
Mechanism mediators from to antibody Antigen- cytokines Acute Weeks (7-21 Cell-Mediated Due to Helper and
mast cells and and Antibody days) Cytotoxic T cell
basophils complement complexes
activation when
Examples Atopic Allergy: HDN, HTR, Serum Contact dermatitis
Hay fever, AIHA, DIHA Sickness, (Rubber, Nickel, they recognize
Asthma, (Penicillin) Arthus Poison Ivy, MHC proteins as
Hives/Urticaria Organ specific Reaction, Cosmetics); foreign
, Angioedema; autoimmune SLE, Tuberculin test Chronic Months to Cell-Mediated Associated with a
Atopic diseases: Dengue, (Mantoux- years notable increase
Dermatitis/Ecz Goodpasture, Post strep Intracutaneous- (>3months) in levels of non T
ema; Hashimoto, GN, RA gold standard for
cell derived non-
Rhinitis (Most Myasthenia Tb, Purified Protein
common gravis, IDDM, Derivative, Von specific growth
atopy/Allergy); Wegeners pirquet-Scratch, factors
Insect sting as Granulomatosi Volmer-Patch- ***This table was adapted and modified from Turqueza’s, Navarro’s and
in bee sting s; standard for Stanley’s
Thrombocytop contact dermatitis);
enia Pneumonitis/Farme
r’s lung/Pigeon SIDENOTES:
breeder’s
disease/Humidifier ➢ Graft-versus-host-disease is a condition that results from
lung disease; transplantation of immunocompetent cells into an immunodeficient
Schistosomiasis, host. The transfused cells attack the tissues of the recipient within
Leprosy, Celiac
disease, Crohn’s the first 100 days post-transplant. Most common with bone marrow
disease, transplantation; Donor cells attack recipient tissue; Key players are
sarcoidosis T cells
➢ Prevention of GVHD:
TRANSPLANTATION IMMUNOLOGY 1. Delete all T cells and supplement graft with T cell derived
1. Transplanted Organs cytokines
-Bone Marrow, Skin, Islets of Langerhans, Heart, Kidney, Liver, 2. Immunosuppressive treatment: Cyclosporin A, FK506
Bone, Xenogeneic Valve Replacements, Cornea (Tacrolimus)
2. Types of Graft 3. Irradiation
Table 21.Types of Graft
GRAFT SOURCE EXAMPLE CD MARKERS
Autograft Tissue is from the Autologous skin Table 23.CD Markers
individual grafting ANTIGEN CELL TYPE FUNCTION
himself/herself CD2 Thymocytes, T cells, NK cells Involved in T cell proliferation
Syngraft/Isograft Tissue is from Kidney transplant, and activation
genetically identical bone marrow CD3 Thymocytes, T cells Associated with T cell antigen
individual (Identical transplant receptor; role in TCR
twins) transduction
Allograft Tissue is from Most organ CD4 Helper T cells, Monocytes Co-receptor for MHC class II;
genetically distinct transplants: and Macrophages Receptor for HIV
(Different) Kidney, heart, CD5 Mature T cells, Thymocytes, Positive or negative
individuals of the lungs etc Subset of B cells (B1) modulation of T and B cell
receptor
same species Fetus on mother’s
CD8 Thymocyte subsets, Co-receptor for MHC class I
womb
Cytotoxic T cells
Xenograft/Heterograft Tissue from Baboon heart
CD10 B and T cell precursors, Protease; marker for pre-B
different transplanted into a Bone marrow stromal cells CALLA
species(e.g. animal) human CD16 Macrophages, NK cells, Low affinity Fc receptor,
Neutrophils Mediates phagocytosis and
ADCC
CD19 B cells, Follicular dendritic Part of B cell co-receptor,
cells, signal transduction molecule
that regulates B cell
development and activation
CD21 Subset of Immature Receptor for complement
thymocytes component C3d; Part of B cell
co-receptor with CD19
3. Graft Rejection CD23 B cells, Monocytes, Follicular Regulation of IgE synthesis;
Table 22.Classification of Graft Rejection Dendritic cells Triggers release of GM-CSF
TYPE TIME FRAME PREDOMINANT CAUSE from monocytes
MECHANISM CD25 Activated T, B cells, Receptor for IL-2
Hyperacute Minutes to Humoral-Mediated Preformed Monocytes
Hours cytotoxic CD44 Most Leukocytes Adhesion molecule mediating
Antibodies to homing to peripheral lymphoid
donor Antigens organs

14 MARCJAYGAGARIN, RMT, MD :3 “A human’s belief can make the impossible, possible. “ –Akihisa Yoshi
IMMUNOLOGY DOCKIE NOTES
CD45R Different forms on all Essential in T and b cell Oncogene HER-2/neu Breast Cancer
hematopoietic cells antigen-stimulated activation Predictive marker for
CD56 NK cells, Subsets of T cells Not known response to
CD94 NK cells, Subsets of T cells Subunit of NKG2-A complex Herceptin/Trastuzumab
involved in inhibition of NK cell therapy
cytotoxicity Expression of this
***This table was adapted from Navarro’s indicates poor prognosis
Others HMB-45 Melanoma
TUMOR MARKERS (Human Melanoma
black 45)
1. TERMINOLOGIES MAGE
(Melanoma
a. Benign Tumor
Associated
-Tumor that does not invade surrounding tissue and normal
Antigen)
body function is preserved
S-100 Melanoma,
b. Malignant Tumor Neuroendocrine Tumors)
-Tumor that continues to grow and can invade surrounding HE4
tissues and greatly disrupts normal body function (Human Ovarian Cancer
-Malignant cells typically differ from visually from normal cells, Epididymis Protein
are metabolically more active to support their growth, and 4)
express different genes or different levels of gene products Cathepsin-D Breast Cancer
as compared to normal cells
Beta-2- Lymphoma
c. Proto-oncogene
microglobulin
-Central regulators of the growth in normal cells that code for
NRLU-10 (Ab Small Cell Lung Cancer
proteins involved in growth and repair processes in the body name)
d. Oncogene
-Product of genetic mutations in proto-oncogenes
3. LAB DETERMINATION
-Oncogen activation causes overexpression of growth
-RIA, IFA, EIA (EIA-most common)
promoting proteins, result in in hypercellular proliferation and
tumorigenesis
AUTOIMMUNE DISEASES
e. Oncofetal Antigens
-Conditions in which damage to organs or tissues results from the presence of
-Antigens that are expressed in the developing fetus and in
autoantibody or autoreactive cells
rapidly dividing tissue, such as that associated with tumors,
but that are absent in normal adult tissue
1. Autoimmune Diseases
Table 25.Spectrum of Autoimmune Diseases
2. TUMOR MARKERS
Table 24.Categories of Protein/Antigen Tumor Markers
TUMOR MARKER EXAMPLES DISEASE
CLASS ASSOCIATIONS
Cell Surface Markers ER/PR Breast Cancer
CD Markers on WBC Neoplasm
WBCs
Immunoglobulins Bence Jones Multiple Myeloma
(Protein) Protein
(Ig Light Chains)
Other Proteins NMP-22, BTA Bladder Cancer
(bladder tumor-
associated
antigen)/CFHrp
(Complement
Factor H-related
protein) ORGAN-SPECIFIC AUTOIMMUNE DISEASES
Oncofetal Antigens AFP Germ Cell Carcinoma
Hepatocellular Carcinoma
CEA Colorectal Cancer
Carbohydrate Antigens CA 125, CA 19-9 Ovarian Cancer
CA15-3, CA 27.29, Breast Cancer
CA 50
CD25 (Il-2 Hairy Cell Leukemia
Receptor)
CD20 Lymphoma
CD45 Hematopoietic
Malignancies
Blood Group Antigens CA 19-9 (Related Pancreatic Cancer
to Lewis Ag)
Enzymes/Isoenzymes PSA Prostate Cancer
ALP Bone and Liver Cancer
***Placental ALP Lung Cancer
AMS Pancreatic Cancer
LDH Lymphoma
GGT Hepatoma
NSE (Neuron Neural Tissue Neoplasms
Specific Enolase)
Hormones hCG Germ Cell Carcinoma
Trophoblastic Carcinoma
Calcitonin Medullary Thyroid Cancer
Gastrin Pancreatic Gastrinoma
SYSTEMIC AUTOIMMUNE DISEASES
15 MARCJAYGAGARIN, RMT, MD :3 “A human’s belief can make the impossible, possible. “ –Akihisa Yoshi
IMMUNOLOGY DOCKIE NOTES
160 is generally considered to be clinically
significant.
-HeLa is an immortal cell line used in scientific
research. It is the oldest and most commonly used
human cell line. The line was derived from cervical
cancer cells taken on February 8, 1951 from
Henrietta Lacks, a patient who died of cancer on
October 4, 1951.
-George Otto Gey found that they could be kept
alive, and isolated one specific cell, multiplied it,
and developed a cell line. Previously, cells
cultured from other human cells would only
survive for a few days.

Table 28.ANA Staining Patterns

Homogenous -Characterizes anti-deoxyribonucleoprotein
(Solid, Diffuse) antibodies (i.e.,antibodies to nDNA, DsDNA,
ssDNA, DNP or histones
-This pattern is typically seen in rheumatoid
disorders
-High titers of homogenous ANA are
suggestive of SLE
-Low titers may be found in SLE, RA,
Sjogren’s syndrome and MCTD
Peripheral -The pattern results from antibodies to DNA
(Ring, Membranous, -Associated with SLE in the active stage of the
Shaggy or Thread) disease and in Sjogren’s syndrome
Speckled -Pattern occurs in the presence of antibody to
(Mottled/Pepper dot) any extractable nuclear antigen devoid of
DNA or histone
-Anti-Sm antibodies have been shown to be
specific with SLE
-Anti-RNP has been found in patients with
wide variety of rheumatic diseases including
SLE, RA, Sjogren’s Syndrome, Progressive
Systemic Sclerosis, MCTD and
Dermatomyositis
Nucleolar -Reflects an antibody to nucleolar RNA
-Present in about 50% patients with
scleroderma (Progressive Systemic
Sclerosis), Sjogren’s Syndrome and in SLE
Anti-centromere -Antibody reacts with the centromeric
Antibody chromatin of metaphase and interphase cells
-Appears to be highly selective for the CREST
variant of progressive systemic sclerosis

3. Anticytoplasmic Antibodies
a. Anti-smooth muscle antibody (anti-SMA)
b. Anti-mitochondrial antibody (AMA)

2. Antinuclear Antibodies
-ANAs are a heterogenous group of circulating immunoglobulins
that include IgG, IgA and IgM. These immunoglobulins react with
the whole nucleus or nuclear components (e.g.,proteins, DNA,
histones) in host tissues
-Present in more than 95% of SLE patients; Present in 4% of
healthy people
-Not specific for SLE since they are also found in other dieases
a. LABORATORY TESTING OF ANA
• Fluorescent Antinuclear Antibody (FANA)
-An example of Indirect Immunofluorescence
-Most widely used and accepted test
-Mouse Kidney or Human Epithelial HEp-2 cells
are fixed to a slide and allowed to react with
patient serum. After careful washing to remove all
unreacted antibody, an antihuman
immunoglobulin with a fluorescent tag or enzyme
label such as horseradish peroxidase is added.
Fluorescent staining pattern is then examined.
The slide is mounted and viewed under a
fluorescent microscope using 400X magnification
(40X objective and 10X eyepiece)
-The screening test is commonly performed with a
1:40 or 1:80 dilution of patient serum in order to
avoid detecting low positive titers that may be
seen in healthy persons, although the exact
dilution used for screening may vary with the
laboratory and population being tested. A titer of

16 MARCJAYGAGARIN, RMT, MD :3 “A human’s belief can make the impossible, possible. “ –Akihisa Yoshi
IMMUNOLOGY
IMMUNODEFICIENCY DISORDERS
-Failure of the immune system to protect against disease or malignancy
1. Primary Immune Deficiencies
-Caused by genetic or developmental defects in the immune
system
-These defects are present at birth but may show up later on in
life
Table 29.Primary Immune Deficiencies
HUMORAL DEFICIENCIES
DISEASE
Bruton X-linked
Agammaglobulinemia
Selective IgA deficiency
Isolated IgG subclass
deficiency
Ataxia Telangiectasia
(Louis-Bar syndrome)
Transient
Hypogammaglobulinemia of
infancy
CELLULAR IMMUNE DEFICIENCIES
DiGeorge Syndrome
(Congenital Thymic
Hypoplasia)
HUMORAL AND CELLULAR DEFICIENCIES
Severe Combined Immune
Deficiency (SCID)
Nezelof Syndrome
Wiskott Aldrich Syndrome
(WAS)
Duncan’s Syndrome
(X-linked Lymphoproliferative
Disease)
DOCKIE NOTES
IMMUNOPROLIFERATIVE DISORDERS
1. Polyclonal Gammopathy
-Tremendous amounts of several classes of immunoglobulins to
several specific antigens are produced, resulting in a broad spike
in the gamma region on serum protein electrophoresis
a. Infectious Diseases
b. Inflammation
c. Liver disease
DEFECIENCY/ CLINICAL FEATURES/ 2. Monoclonal Gammopathy
DEFECT DESCRIPTION
All classes of Igs -Genetic defect in long arm of -Malignant transformation of a clone of B cells that produce
chromosome X resulting in a block identical antibodies
in the maturation of pre B cells into a. Multiple Myeloma
lymphocytes with surface IgM
leading to a complete absence of B
b. Waldenstrom’s Macroglobulinemia
cells and plasma ells in the
circulation
IgA -Usually caused by a genetic
defect or by drugs (phenytoin and
penicillin)
-Caused by impaired differentiation
of lymphocytes to become IgA-
producing plasma cells
-Most common primary immune
deficiency
-Causes Anaphylaxis when
administered or transfused to
someone with Selective IgA
deficiency
IgG -Most antibodies directed against
IgG4: Most protein antigens are of the IgG1
common and IgG3 subclasses
IgG1: Least -Those directed against
common carbohydrate antigens are of the
IgG2 and IgG4 subclasses
IgA and IgE -Ataxia: Uncoordinated muscle
movement especially on the
earlobes and conjuctiva
Telangiectasia: Dilation of small
blood vessels on the earlobes and
conjunctiva
Igs Many infants go through a period of
(Particularly IgG) hypogammaglobulinemia between
the 5th-6th month of life
T cells -Faulty development of 3rd and 4th
pharyngeal pouches during
embryogenesis
-Aplasia or hypoplasia of thymus
and parathyroid glands
-Associated with microsomal
deletion on chromosome 22
-Abnormally high CD4+/CD8+
ratio is present because of a
decrease in CD8+ cells
Igs and T cells 2 forms:
a. Autosomal
Recessive Form
-ADA deficiency
b. X-linked Form
-Mutation in the
gene encoding the
IL-2 receptor gamma
chain
Igs and T cells -Thymic dysplasia
-Primary Immunodeficiency most
commonly confused with AIDS in
the pediatric group
IgM -Mutation in WASP, a protein
Motility of T cells involved with cytoskeletal
are defective reorganization necessary for
delivering cytokines
-Characterized by a triad of:
Eczema, Thrombocytopenia,
Recurrent Infections
-Platelets are small and defective
-Absence of isohemagglutinins is
the most consistent lab finding and
is often used diagnostically
Weak humoral and -Mutation in SH2DIA/SAP gene
cell-mediated -Particularly vulnerable to EBV
immune response infection

2. Secondary Immune Deficiencies (Acquired)

-Loss of immune function as a result of exposure to disease
agents, environmental factors, immunosuppression or aging
a. MALIGNANCY
-Cancers, Lymphoma, CLL, Multiple Myeloma
b. VIRAL DISEASE
- HIV, EBV, CMV
c. NUTRITIONAL DEFICIENCY
-Marasmus

17 MARCJAYGAGARIN, RMT, MD :3 “A human’s belief can make the impossible, possible. “ –Akihisa Yoshi