Immunology & Serology

IMMUNOLOGY : study of cells, tissues and organs responsible for the recognition and disposal of non-self antigens

SEROLOGY : study of the constituents of serum

HISTORICAL PERSPECTIVE
 Individuals who recovered from certain infectious diseases were thereafter protected from the disease
 430 BC : Thucydides : after a plague in Athens he wrote that only those who had recovered could nurse the sick
because they would not contract the disease a second time
 15th century : Chinese and Turks : dried crusts from smallpox pustules were either inhaled or inserted into
small cuts in the skin (VARIOLATION)
 1718 : Lady Mary Montagu ; variolation of her own children
 1798 : Edward Jenner : cross-immunity using cowpox to protect against smallpox
 Christopher Columbus
 Louis Pasteur : hypothesized and proved that aging had weakened the virulence of the pathogen hence may be
used to protect against the disease
 Vaccine – attenuated strain
 1885 : administered 1st vaccine to a human boy (rabies vaccine)
 1890 : Emil von Behring and Shibasaburo Kitasato : mechanism of immunity : demonstrated that serum from
animals previously immunized against diphtheria could transfer the immune state to unimmunized animals
 1901 : Emil von Behring received the Nobel Prize for his discovery of serum antitoxins
 1902: Portier and Richet: immediate hypersensitivity reaction
 1905 : Robert Koch received the Nobel Prize for his discovery of cellular immunity in TB
 1930 : Elvin Kabat : immunoglobulin called antibodies contained in body fluids are protective agents (HMI)
 1883 : Elie Metchnikoff : certain WBC (phagocytes) were able to ingest microorganisms and foreign material (CMI)
 1908 : Elie Metchnikoff received the Nobel Prize for his discovery of phagocytosis
Paul Ehrlich received the Nobel Prize for his studies on immunity
 1913 : Charles Richet received the Nobel Prize for his studies on anaphylaxis
 1919 : Jules Bordet received the Nobel Prize for his discovery of the complement cascade
 1930 : Karl Landsteiner received the Nobel Prize for his discovery of the human blood group antigens (ABO)
 1949: Salk and Sabin: polio vaccine
 1950 : lymphocyte : cell responsible for both HMI and CMI
 1951: Reed: yellow fever vaccine
 1960 : Macfarlane Burnet and Peter Medawar received the Nobel Prize for their discovery of immunologic
tolerance
 1972 : Gerald Edelman and Rodney Porter received the Nobel Prize for their discovery of the antibody
structure
 1977 : Rosalyn Yallow received the Nobel Prize for developing the radioimmunoassay (RIA) technique
 1980 : George Snell, Jean Dausset, Baruj Benaceraf received the Nobel Prize for their discovery of the major
histocompatibility complex (MHC)
 1984 : Niels Jerne received the Nobel Prize for his discovery of immunoregulation
Georges Koehler, Cesar Milstein received the Nobel Prize for developing monoclonal antibody production
technique
 1984 : year of discovery of the T cell receptor gene
 1987 : Susumu Tonegawa received the Nobel Prize for his discovery of antibody diversity
 Frazer: human papilloma virus
 Bruce Glick : CMI : T lymphocytes (thymus derived) and HMI : B lymphocytes (Bursa of Fabricius)
 1991 : Edward Donnall Thomas, Joseph Murray received the Nobel Prize for their study on transplantation
 1996 : Peter Doherty, Rolf Zinkernagel received the Nobel Prize for their study on cytotoxic T cell recognition of
virally infected cells
 2008 : Françoise Barré-Sinoussi, Luc Montagnier received the Nobel Prize for their discovery of the human
immunodeficiency virus (HIV)

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OVERVIEW OF THE IMMUNE SYSTEM

 Versatile defense system : protects against invading pathogenic microorganisms and cancer
 Immune responses:
 Recognition
 Response

IMMUNE SYSTEM: INNATE AND ADAPTIVE COMPONENTS

 Immunity – state of protection from infectious disease
 Innate – less specific; broad reactivity
 Disease-resistance mechanisms not specific to a particular pathogen
 Adaptive – more specific
 Antigen exposure is a prerequisite
 High degree of specificity
 Immunologic memory

INNATE IMMUNITY
External Defense System
 Unbroken Skin and Mucous Membranes
 Major anatomic and physical barrier
 Several secretions present discourages the growth of microorganisms
 Mucous secretion
 Lysozyme
 Normal flora keeps pathogens from establishing themselves
 Low pH
 Special surface structures

Internal Defense System

HUMORAL COMPONENTS
 Complement
 Interferon (IFN)
 Type 1: IFN-α

IFN-β

 Type 2: IFN-γ

 Beta-lysin

 Tumor Necrosis Factor (TNF)

 TNF-α

 TNF-β

 Acute Phase Reactants

 PHAGOCYTOSIS
1. Initiation : phagocytosis is initiated as a result of tissue damage
 results to increased surface adherence receptors
 CR3,
 Laminin receptor
 N-formyl-methionyl-leucylphenylalanine receptor

2. Chemotaxis : movement of cells towards the foreign entity

 cells tend to move in a certain direction under the stimulation of a chemical substance
 Neutrophils : first to migrate but short-lived
 Monocytes : second to migrate but long-lived
 Positive Chemotaxis: phagocytes move toward the site of stimulation
 Negative Chemotaxis: phagocytes move away from the site of stimulation
 Job’s Syndrome
 Lazy Leukocyte Syndrome
3. Attachment :
 Direct Interaction
 PAMPs : Pathogen-Associated Molecular Patterns
 PRRs : Pattern Recognition Receptors
1. PRRs in blood and lymph (complement  opsonization)
2. Phagocytic receptors (pathogen binding  release of effector molecules)
3. TLRs : Toll-Like Receptors
 TLR1  lipoprotein
 TLR2  teichoic acid and peptidoglycan
 TLR4  lipopolysaccharide
 Indirect Interaction
 Through OPSONIZATION

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 Opsonins
 Complement factors (C3b, C4b, C5b)
 Antibodies (IgG1 and IgA)
 Acute Phase Reactants

4. Engulfment :
 facilitated by opsonization : coating of foreign entity with plasma factors
 phagosome formation

5. Digestion:
 Mechanisms in Antigen Destruction
A. Oxygen-Dependent Mechanism
 involves increase in oxygen consumption and stimulation of the Hexose
Monophosphate Shunt
 generation of bactericidal metabolites

B. Oxygen-Independent Mechanism
 Production of nitric oxide
 Phagolysosome formation

 INFLAMMATION
 Tissue damage and infection induce leakage of vascular fluid, containing serum proteins with
antibacterial activity, and influx of phagocytic cells into the affected area
 Cardinal signs
1. Rubor: increased blood flow to the area
2. Tumor: leakage of blood plasma to surrounding tissues
3. Calor: IL 1 secreted by mononuclear phagocytes
4. Dolor: lactic acid production by phagocytes
5. Functio laesa : loss of function

Major Events in an Inflammatory Response

VASCULAR RESPONSE
1. Vasodilation
 Increase in diameter of blood vessels
2. Increase in capillary permeability
 Influx of fluid and cells into the tissue

CELLULAR RESPONSE
1. Influx of phagocytes
 Neutrophils
 First to migrate within 30 to 60 minutes and emigration lasts from 24 to 48 hours
 Monocytes/Macrophages
 Second to migrate within 4 hours and peaks at 16 to 48 hours
 Secrete soluble substances
 Humoral Response

RESOLUTION AND REPAIR

 initiated by fibroblasts

 CELLS THAT FUNCTION IN INNATE IMMUNE RESPONSES

A. PHAGOCYTES : main role is to engulf and destroy foreign substances
1. Neutrophils
 polymorphonuclear neutrophilic leukocyte that constitutes 50 to 70 % of the total peripheral
WBC population
 10-15um with nucleus containing 2-5 lobes

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 marginating pool and circulating pool

2. Monocytes
 mononuclear cells that constitute 4 to 10 % of circulating WBCs
 largest cells in the peripheral blood with a diameter of 12 to 22 um and an irregularly folded or
horseshoe-shaped nucleus
3. Tissue Macrophages
 25 to 80 um in size
 part of the monocyte-macrophage system that initiates and regulates immune response
 play a role different immune responses: microbial killing, tumoricidal activity, intracellular
parasite eradication, phagocytosis, secretion of cell mediators, and antigen presentation

NAME LOCATION
Lungs
Connective Tissues
Liver
Kidneys
Brain
Bone
Spleen
Placenta
Synovium
Blood
Skin
Lymph Nodes

B. NATURAL KILLER CELLS

 main role is destruction of abnormal autologous cells
 ability is enhanced by the presence of cytokines

C. INFLAMMATORY CELLS
1. Basophils
 less than 1% of circulating WBCs
 10-15 um
 contain granules that induce and maintain immediate hypersensitivity reactions
2. Eosinophils
 constitutes 1 to 3 % of circulating WBCs
 12-15 um in diameter with bilobed or ellipsoidal nucleus
 neutralizes basophil and mast cell products and killing certain parasites
3. Mast Cells
 connective tissue cells of mesenchymal origin
 larger than basophils and contain more granules
 life span is between 9 to 18 months
 plays a role in hypersensitivity reactions

D. ANTIGEN PRESENTING CELLS

1. Dendritic Cells
 main function is to process and present antigens to T helper lymphocytes
 most potent phagocytic cell in tissues

COMPLEMENT
Major Functions
1. OPSONIZATION
 Coating of pathogenic organisms or immune complexes prior to phagocytosis
2. INFLAMMATION
 Induction of histamine release from mast cells and basophils
3. CYTOLYTIC FUNCTION
 Membrane attack of target cells leading to cell lysis/death

PROTEINS OF THE COMPLEMENT CASCADE

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 to form C3 convertase
C3 Key intermediate in all pathways

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C5 Initiates membrane attack complex

C6 Binds to C5b in MAC
C7 Binds to C5b6 in MAC
C8 Starts pore formation on membrane of target cell
C9 Polymerizes to cause cell lysis
ALTERNATIVE PATHWAY
Factor B Binds to C3b to form C3 convertase
Factor D Cleaves factor B
Properdin Stabilizes C3 convertase (C3bBb)
MBL PATHWAY
MBL Binds to mannose
MASP-1 Helps cleave C4 and C2
MASP-2 Cleaves C4 and C2

CLASSICAL ALTERNATIVE MBL

PLASMA COMPLEMENT REGULATION

C1 Inhibitor (C1INH) Dissociates C1r and C1s from C1q
Factor I Cleaves C3b and C4b
Cofactor to factor I in the inactivation of C3b
Factor H
Prevents binding of B to c3b
C4-binding protein Cofactor to factor I in the inactivation of C4b
S protein Prevents attachment of C5b67 complex to cell membranes

DEFICIENCIES OF COMPLEMENT COMPONENTS

Lupus-like syndrome
C1(q, r, or s)
Recurrent infections

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Lupus-like syndrome
C2 Recurrent infections
Atherosclerosis
Severe recurrent infections
C3
Glomerulonephritis
C4 Lupus-like syndrome
C5-C8 Neisseria infections
C9 No known disease association
C1INH Hereditary angioedema
DAF
Paroxysmal nocturnal hemoglobinuria
MIRL/CD59
Factor H or Factor I Recurrent pyogenic infections
Pneumococcal diseases
MBL Sepsis
Neisseria infections
Properdin Neisseria infections
MASP-2 Pneumococcal diseases

ADAPTIVE IMMUNITY
Characteristics of Adaptive Immunity
1. Antigenic specificity – can distinguish between 2 protein molecules that differ in only a single amino acid
2. Diversity – capable of recognizing a wide variety of antigens
3. Immunologic memory – second encounter with the same antigen induces a heightened immune response
4. Self/non-self recognition – capable of distinguishing self from non-self antigens

Types of Adaptive Immunity

1. Cell-Mediated Adaptive Immunity
2. Humoral-Mediated Adaptive Immunity
Involvement of the Body in Manner of Administering Examples
Antibody Production Immunogen or Antibody
NATURAL ACTIVE Infection
PASSIVE Breast feeding
Transplacental transfer
ARTIFICIAL ACTIVE Immunization
PASSIVE Administration of serum
Immunoglobulins

LYMPHOID ORGANS
TWO MAIN GROUPS:
1. Primary Lymphoid Organs
 Provide appropriate microenvironments for the development and maturation of lymphocytes
 Releases mature lymphocytes
 Thymus and bone marrow
2. Secondary Lymphoid Organs
 Trap antigen from defined tissues or vascular spaces and are sites where mature lymphocytes
can interact effectively with the antigen
 Site for antigen-dependent lymphopoiesis
 Lymph nodes, spleen, MALT : GALT

PRIMARY LYMPHOID ORGANS

BONE MARROW
 Largest lymphoid organ (1300 to 1500 g)
 Center for antigen-independent lymphopoiesis
 Primary source of pluripotent stem cells that give rise to all HSC
 Major organ for B cell maturation and gives rise to the precursor cells of the thymic lymphocytes
 Functions :
1. Produces large numbers of B cells, each with unique antigen receptors (antibodies) such that, overall,
there is sufficient B cell diversity to recognize millions of microbial antigens in the environment
2. Eliminates B cells with antigen receptors having high affinity for self-molecules

THYMUS
 Site of T-cell development and maturation
 Flat bilobed organ situated above the heart
 Each lobe is surrounded by a capsule and divided into lobules
 Each lobule is composed of the following areas/regions:
1. Outer cortex : densely packed with immature T cells : thymocytes
2. Inner medulla : sparsely populated with thymocytes
 Progenitor T cells appear as early as 8 weeks of gestation
Thymocytes  Cortex  Medulla  Mature T cells are released

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SECONDARY LYMPHOID ORGANS

SPLEEN
 Large, encapsulated bean-shaped organ with a spongy interior situated on the left side of the body below the
diaphragm
 Largest secondary lymphoid organ
 Filters blood by trapping blood borne microorganisms and producing an immune response to them
 Removes damaged RBCs and immune complexes
 Reservoir of RBCs and platelets
 Divided into:
1. Red Pulp
 Consists majority of splenic tissue
 Contains RBCs and numerous arterioles
 Carries out hematologic function of the spleen
2. White Pulp
 Consists approximately 20% of splenic tissue
 Contains white blood cells and lymphoid tissue
 PALS(periarteriolar lymphoid sheath)
 Lymphoid tissue arranged around arterioles
 Contains T cells
 Primary Follicles
 Contains unstimulated B cells
 Germinal Centers
 Where stimulated B cells migrate to
 Marginal Zone
 Area that surround PALS
 Contains dendritic cells
 Splenectomy : increases risk of infection with encapsulated bacteria and severe malaria

LYMPH NODES
 Small solid structures found at varying points along the lymphatic system
 2-10mm in size, spherical and possess an enveloping capsule
 Filters lymph and then produces an immune response against any microbe they trap
 Divided into 3 parts or areas:
1. Outer Cortex
 Contains macrophages
 Primary Follicles
 Mature, resting B cells
 Follicular Dendritic cells
 Secondary Follicles
 Contains Ag-stimulated proliferating B cells
 Plasma cells: produce and secrete antibodies
 Memory B cells
2. Paracortex
 Region between follicles and medulla
 Contains T lymphocytes in close proximity to interdigitating cells
3. Medulla
 Less densely populated with T cells and B cells
 Contains macrophages and numerous plasma cells
 Transit time through a lymph node: 18 hours
 In the presence of Ag contact lymphocyte traffic shuts down

OTHER LYMPHOID TISSUES

MUCOSA-ASSOCIATED LYMPHOID TISSUE (MALT)
 50% of lymphoid tissue in the human body is located lining the major tracts : respiratory, digestive and
genitourinary

Four related but distinct cell membrane molecules are responsible for antigen recognition by the immune system:
1. Membrane-bound antibodies on B cells
2. T-cell receptors
3. CD8 Receptor for Class I MHC molecules
4. CD4 Receptor for Class II MHC molecules

CELLS INVOLVED IN ADAPTIVE IMMUNITY

T LYMPHOCYTES
 E-Rosette Assay: uses reagent sheep RBC
 Represent ≈ 80% of lymphocytes in the circulation
 Surface markers of circulating T lymphocytes include:
 CD2: sheep RBC receptor
 CD3: forms part of the TCR
 CD4: receptor for MHC Class II molecule
 CD8: receptor for MHC Class I molecule
 T cell subsets or populations include:

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 T helper cells: CD4+

 Inducer T cells: CD4+
 Cytotoxic T cells: CD8+
 Suppressor T cells: CD8+
T CELL ONTOGENY
 THYMOCYTES: lymphocyte precursors produce by the bone marrow
 Early thymocyte markers: CD44 and CD25

1. DOUBLE-NEGATIVE STAGE
 Lacks CD4 and CD8 surface markers
 Thymic stromal cells: epithelial cells, macrophages, fibroblasts and dendritic cells
 IL 7: for growth and differentiation
 Rearrangement of genes coding for Ag receptor
 Antigen receptor: T CELL RECEPTOR (TCR)
 CD3
 8 chains common to all T cells
 2 chains responsible for variability
 Rearrangement of beta chains occurs first
 Beta chain signal: triggers thymocytes to express surface markers
**Other thymocytes: express gamma and delta
become T cell population in skin, intestinal epithelium and pulmonary epithelium
 Additional Markers: CD2, CD5 and CD7

2. DOUBLE-POSITIVE STAGE
 Thymocytes have CD4 and CD8 markers
 Begin rearrangement for alpha chains
 Complete TCR expressed on the surface
 SELECTION PROCESS:
 POSITIVE SELECTION
(+) functional TCR to survive
(+) ability to recognize foreign antigens in association with MHC I and II
**inability to recognize self-MHC presented antigens  DEATH
 NEGATIVE SELECTION
(-) strong reactions with self-antigens
**ability to react with self-antigens  APOPTOSIS

3. SINGLE-POSITIVE STAGE
 T cells exhibit either CD4 or CD8 surface marker
 NEGATIVE SELECTION: if cells are reactive to self-antigens  DEATH
 CD4 cells: recognize antigens presented via MHC II
 T helper cells (Th)
A. Th 1: intracellular pathogens  cytotoxic T cell activation
 produce interferon gamma and tumor necrosis factor beta
B. Th 2: extracellular pathogens  B cell activation
 produce interleukins (IL 4, IL 5, IL 10 and IL 13)
 T regulatory cells (Treg)
 Contains CD4 and CD25 surface markers
 Produces IL 10 and transforming growth factor B
 Suppress immune response to self-anitgens
 CD8 cells: recognize antigens presented via MHC I

4. ACTIVATED T CELL
 Requires interaction with an antigen
 Antigen interaction  T cells transform and express IL 2 receptors
 T lymphoblasts  differentiate to active lymphocytes
 Effector cells: cytotoxic T cells (Tc)
 Memory T cells: persist for years and produce a broader array of cytokines
 Cell-mediated immunity starts:
 Commencement of Ab production by B cells
 Killing of tumor or other target cells
 Rejecting grafts
 Stimulation of hematopoiesis
 Initiation of delayed hypersensitivity reaction

B LYMPHOCYTES
 EAC Rosette Assay
B CELL ONTOGENY
 B cells remain in the microenvironments provided by bone marrow stromal cells
 GROWTH FACTORS: E2A, Early B cell Factor (EBF), Paired Box Protein 5 (PAX)
 CYTOKINE: Interleukin 7 (IL 7)

1. PRO-B CELLS
 Rearrangement of genes on chromosome 14

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 C-KIT interacts with stem cell factor

 RAG1 and RAG2 cleave DNA at recombination sites
 TdT helps join the pieces back together by incorporating additional nucleotides
 Surface markers present: CD19, CD45R, CD43, CD24, C-Kit
2. PRE-B CELLS
 Synthesis of heavy chains occurs (mu heavy chain)
 Mu chains accumulate in the cytoplasm
 Loss of surface markers
 Formation of pre-B cell receptor

3. IMMATURE B CELLS
 Appearance of complete IgM molecules on the surface
 Rearrangement of genes for light chains
 Completion of light chain rearrangement
 Mu chains in the cytoplasm are no longer detectable
 Surface Markers: CD21, CD40, MHC Class II

4. MATURE B CELLS
 Marginal Zone B cells: remain in the spleen to provide a quick response to blood-borne pathogens
 Follicular B cells: found in lymph nodes and other tissues

5. ACTIVATED B CELLS
 Activation Site: primary follicles of lymphoid tissues and organs
 Expression of CD25

6. PLASMA CELLS AND MEMORY B CELLS

 Characterized by presence of cytoplasmic Ab rather than surface Ab
 Found in germinal centers of peripheral lymphoid organs
 Memory Cells: longer lifespan than resting B cells
capable of responding to Ag with increased speed and intensity

MITOGENS FOR LYMPHOCYTES

 T CELLS: Concanavalin A, Phytohemagglutinin, Pokeweed Mitogen
 B CELLS: Lipopolysaccharides, Pokeweed Mitogen

THIRD POPULATION LYMPHOCYTES

 Mediate cytolytic reactions and kill without prior exposure
 Function as transitional cells bridging innate and acquired immunity
 IL 15: plays a critical role for NK developmet
 Locations:
 Maternal-fetal interface
 Peripheral blood
 Spleen
 Liver
 NK cell activity is stimulated by IL 12, IFN gamma, IFN beta

MECHANISM OF NK CELL ACTIVITY

1. Ab-INDEPENDENT CYTOTOXICITY
 Response governed by receptors
 Inhibitory Receptors
 recognize MHC Class I on the surface of healthy cells
 recognition is facilitated by KIRs, ILT/LIR, CD94/NKG2A

 Activating Receptors
 diseased and cancerous cells lose the ability to produce MHC proteins
 CD94/NKG2B and CD94/NKG2C
 MICA and MICB: proteins expressed by stressed cells
 Two Substances for Destruction
 Perforins: pore-forming substance that acts on abnormal cells in the presence of
calcium
 Granzymes: packets of serine esterases that mediate cell lysis

2. Ab-DEPENDENT CYTOTOXICITY
 NK cells recognize and lyse Ab-coated cells
 CD 16: receptor for the Fc portion on IgG

IMMUNOGENS AND ANTIGENS

IMMUNOGENICITY
 Ability to induce a humoral and/or cell-mediated immune response
 IMMUNOGEN  substance capable of inducing an immune response

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ANTIGENICITY
 Ability to combine specifically with the final products of humoral and/or cell-mediated immune response
 Complete antigen: carrier + hapten
 Incomplete antigen: hapten only

FACTORS THAT INFLUENCE IMMUNOGENICITY

A. NATURE OF IMMUNOGEN
1. FOREIGNNESS
 The greater the phylogenetic distance between two species, the greater the structural disparity,
the more immunogenic.
 Autoantigen
 Alloantigen
 Heteroantigen
 Heterophile antigens
**Influences GRAFT REJECTION
Most Immunogenic:
Skin
Islets if Langerhans
Heart
Kidney
Liver
Bone
Xenogeneic Valve Replaacements
Least Immunogenic:

2. MOLECULAR SIZE
 The larger the size, the more immunogenic.
 Potent antigen: MW greater than 10000 Daltons
 Good immunogen: MW greater than 40000 Daltons
 Excellent immunogen: MW greater than 1000000 Daltons

3. CHEMICAL COMPOSITION AND HETEROGENEITY

 The more complex the structural and biochemical make-up, the more immunogenic.
 Protein
 Polysaccharides
 Lipids and Nucleic acids

4. SUSCEPTIBILITY TO ANTIGEN PROCESSING AND PRESENTATION

 The larger and more insoluble, the more immunogenic.

B. BIOLOGICAL SYSTEM
1. GENOTYPE
 Difference in the response due to genetic variability among individuals

2. DOSAGE AND ROUTE

 The higher the dose, the greater the immune response.
 Immune response depends on the route of administration.
 Intravenous
 Intradermal
 Subcutaneous
 Intramuscular
 Intraperitoneal

3. ADJUVANTS
 Substances that enhance immunogenicity
 Examples
 Complete Freund’s Adjuvant (CFA)
 water in oil emulsion (Mycobacterium butyricum, Bordetella pertussis,
Mycobacterium tuberculosis)
 Lipopolysaccharide (LPS)
 Alum adjuvant
 Squalene (MF59)
 Effects
 Antigen persistence is prolonged
 Co-stimulatory signals are enhanced
 Local inflammation is increased
 Non-specific proliferation of lymphocytes is stimulated

EPITOPES
 Immunologically active regions of an immunogen that bind to antigen-specific membrane receptors on
lymphocytes or to secreted antibodies
 B and T cells recognize different epitopes on the same antigenic molecule

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CHARACTERISTIC B CELLS T CELLS

Interaction with antigen
Binding of soluble antigen
Involvement of MHC
molecules
Chemical nature of antigens
Epitope properties
Involves binary complex of
membrane Ig and Ag
Yes
None required
Protein, polysaccharide, lipid
Accessible, hydrophilic, mobile
peptides
Involves ternary complex of T
cell receptor, Ag and MHC
molecule
No
Display of processed antigen
required
Mostly proteins presented on
MHC molecules
Internal linear peptides
bound to MHC molecules

HAPTENS
 Small organic molecules that are antigen but not immunogenic
 Needs a carrier to become accessible to the immune system

MAJOR HISTOCOMPATIBILITY COMPLEX

 Tightly linked cluster of genes whose products play roles in intercellular recognition and in discrimination
between self and nonself
 Participates in the development of both HMI and CMI
 Set of genes that control tissue compatibility in man (HLA)
 MHC genes are polymorphic : large numbers of alleles for each gene
 Collection of genes arrayed within a long continuous stretch of DNA on the SHORT ARM of CHROMOSOME 6
 Organized into three regions
 Class I MHC genes – encode glycoproteins expressed on the surface of nearly all nucleated cells; major
function : presentation of peptide antigens to Tc cells
 HLA A and B: most significant
 HLA C: low concentration
 HLA E and F: not expressed
 HLA G: expressed in trophoblastic cells
 Class II MHC genes – encode glycoproteins expressed primarily on APC; function : present processed
antigenic peptides to Th cells
 Class III MHC genes – encode components of the complement system and molecules involved in
inflammation

ANTIGEN PROCESSING AND PRESENTATION

 T cells recognize antigen displayed within the cleft of a self-MHC molecule on the membrane of a cell
 In general CD4 Th cells recognize antigen with class II MHC molecules on antigen processing cells
 CD8 Tc cells recognize antigen with class I MHC molecules on target cells

Processing of Endogenous Antigens Processing of Exogenous Antigens

IMPORTANCE OF MHC/HLA
1. Disease Association/Susceptibility
 Autoimmune disorders, certain viral diseases, disorders of the complement system, some neurologic disorders
and several different allergies
 Ankylosing spondylitis : B27
 Goodpasture’s syndrome : DR2
 Type 1 DM : DR4/DR3
 Multiple sclerosis : DR2
 SLE : DR3

peeweebells Chance favors those who are prepared. 11

 Immunology & Serology

2. Tissue/Organ Transplant
 Siblings: 50%
 Parents: 25%
3. Paternity Testing

ANTIBODIES
• Glycoproteins present in serum
• Gamma globulins produced by plasma cells

MONOMER: 4 polypeptide chains

1. HEAVY CHAINS
 MW: 50,000 to 75,000
 Approx. 400 AA
 determines Ig class
 with SUBCLASSES: IgG, A, M
2. LIGHT CHAINS
 MW: approx. 23,000
 200-220 AA
 Kappa: 65%
 Lambda: 35%
3. DISULFIDE BONDS: Hold the 4 polypeptide chains together
 INTERCHAIN BONDS: H-H, H-L, L-L
 INTRACHAIN BONDS: stronger and occur within individual chain type
4. HINGE REGION
 Flexible region of the Ig
 AA component: PROLINE
5. DOMAINS
VARIABLE REGION
 first 100 AA in the N-terminal
 contains hypervariable region
CONSTANT REGION
 demonstrates an unvarying AA sequence in the carboxy terminal
LIGHT CHAIN
 2 domains (1VLC + 1CLC)
HEAVY CHAIN
 4 domains (1VHC + 3CHC)
FRAGMENTS
 proteolytic degradation products of Ig molecules
 PAPAIN: attack HINGE region
 PEPSIN: digestion of most part of Fc

OTHER TERMS
• ISOTYPE: HC that determines Ig class
• ALLOTYPE: variation in constant region of HC and LC
• IDIOTYPE: variation in the variable region of HC and LC

STRUCTURE OF Ig CLASSES
• MONOMER
 IgG, IgD, IgE, most serum IgA
• DIMER
 Secretory IgA
• POLYMER
 Serum IgM

PROPERTIES OF IMMUNOGLOBULINS
IMMUNOGLOBULIN GAMMA
 predominant Ig in humans
 4 subclasses: IgG1, 2, 3, 4
 provide immunity to NB except IgG2
 Complement fixation except IgG4
 Best precipitin
IMMUNOGLOBULIN MU
 Most primitive
 Best agglutinin
 Complement fixation

IMMUNOGLOBULIN ALPHA
 IgA1: serum
 IgA2: secretions
 J chain
 Secretory piece/component
 For mucosal immunity
 Prevents attachment of pathogen

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 Immunology & Serology

IMMUNOGLOBULIN DELTA
 Unstimulated, immunocompetent B cells
 Immunoregulation
 anti-idiotypic Ab
IMMUNOGLOBULIN EPSILON
 Heat-labile originally called REAGIN
 High affinity for basophils and mast cells
 Recognition of parasites
 RIST
 RAST

THEORIES ON THE SPECIFICTY OF ANTIBODY-ANTIGEN REACTIONS

1. Selective theory
 Paul Ehrlich 1900
 Cells in the blood express a variety of receptors that could react with infectious agents and inactivate
them
 Infectious agent and a cell bound receptor will induce the cell to produce and release more receptors
with the same specificity
 Specificity of receptor is determined before antigen exposure

2. Instructional theory
 1930(Breinl and Haurowitz) -1940 (Pauling)
 Antigen : determines the specificity of the antibody molecule
 Antibody molecule would then assume a configuration complementary to that of the antigen template
 Antigen will serve as the template around which antibody would fold
3. Clonal Selection Theory
 1950 : Jerne, Talmadge and Burnet
 Individual lymphocyte expresses membrane receptors that are specific for a distinct antigen
 Unique receptor specificity is determined before the lymphocyte is exposed to the antigen
 Binding of antigen to specific receptor activates it to proliferate into a clone of cells with the same
immunologic specificity as the parent cell

KINETICS OF HUMORAL RESPONSE

1. LAG PHASE: no detectable Ab
2. LOG PHASE: Ab titer increases logarithmically
3. PLATEAU PHASE: Ab titer stabilizes
4. DECLINE: Ab is catabolized

PRIMARY RESPONSE SECONDARY RESPONSE

LAG 7-10 days 1-2 days
Slow exponential increase in Ab Ab concentration increases more
LOG
titer rapidly and reaches a higher titer
PLATEAU Short lived Longer
DECLINE More gradual decline
PREDOMINANT Ab IgM IgG
MAGNITUDE OF RESPONSE Stronger response

IMMUNE DYSFUNCTION AND ITS CONSEQUENCES

- Failure to protect the host adequately
- Misdirection of immune activities
- Discomfort, debilitating disease or death

HYPERSENSITIVITY

COMPARISON OF HYPERSENSITIVITY REACTIONS

TYPE I TYPE II TYPE III TYPE IV
Immune Mediator IgE IgG IgG or OgM T Cells
Autologous or Autologous or Autologous or
Antigen Heterologous
Heterologous Heterologous Heterologous
Complement
Involvement
Release of mediators Cytolysis due to
Deposits of antigen-
Immune Mechanism from mast cells and antibody and Release of cytokines
antibody complexes
basophils complement
Anaphylaxis
Transfusion reactions Serum sickness Contact dermatitis
Hay fever
Examples AIHA Arthus reaction Tuberculin test
Food allergies
HDFN SLE Pneumonitis
Asthma

peeweebells Chance favors those who are prepared. 13