IMMUNOLOGY

 Study of the reactions of a host when foreign substances are introduced into the
body
 Study of all aspects of body defenses, such as antigens and antibodies, allergy and
hypersensitivity

NOBEL PRIZE WINNERS IN IMMUNOLOGY

YEAR SCIENTIST RESEARCH
1901 Emil von Behring Serum antitoxins
1905 Robert Koch Cellular immunity in TB
1908 Elie Metchnikoff, Phagocytosis
Paul Ehrlich Immunity
1913 Charles Richet Anaphylaxis
1919 Jules Bordet Complement
1930 Karl Landsteiner Human blood group antigens
1960 Macfarlane Burnet, Peter Discovery of immunologic tolerance
Medawar
1972 Gerald Edelman, Rodney Porter Structure of antibodies
1977 Rosalyn Yalow Radioimmunoassay
1980 George Snell, Jean Dausset, Baruj Major histocompatibility complex
Benaceraf
1984 Niels Jerne, Immunoregulation
Georges Koehler, Cesar Milstein Monoclonal antibody
1987 Susumu Tonegawa Antibody diversity
1991 Edward Donnall Thomas, Joseph Transplantation
Murray
1996 Peter Doherty, Rolf Zinkernagel Cytotoxic T cell recognition of virally
infected cells
2008 Francoise Barré-Sinoussi, Luc Human immunodeficiency virus
Montagnier

SIGNICANT MILESTONES IN IMMUNOLOGY

1798 Jenner Smallpox vaccination 1953 Graft versus host
reaction
1862 Haeckel Phagocytosis 1957 Burnet Clonal selection theory
1880 Pasteur Live, attenuated 1957 Interferon
- chicken cholera and
1881 anthrax vaccines
1883 Metchniko Cellular theory of 1958- HLAs
- ff immunity through 1962
1905 phagocytosis
1890 Von Humoral theory of 1964- T cell and B cell
behring, immunity proposed 1968 cooperation in
kitasata immune response
1891 Koch Demonstration of 1972 Identification of
cutaneous (delayed- antibody molecule
type) hypersensitivity
1900 Ehrlich Antibody formation 1975 Köhler First monoclonal
theory antibodies
1902 Portier, Immediate- 1985- Identification of genes
richet hypersensitivity 1987 for T cell receptor
 anaphylaxis
1903 Arthus 1986 Monoclonal hepatitis B
vaccine
1938 Marrack 1986 Mosman Th1 versus Th2 model
n of T helper cell
function
1944 1996- Identification of toll-
1998 like receptors
1949 Salk, sabin Development of polio 2001 FOXP3, the gene
vaccine directing regulatory T
cell development
1951 Reed Vaccine against yellow 2005 Frazer Development of
fever human papillomavirus
vaccine

HISTORICAL PERSPECTIVES (STANLEY)

1798 Edward Jenner, an English countryside physician demonstrated that
protection from cowpox could be generated by the transfer of postural
material from a cowpox lesion instead of the more hazardous smallpox
lesion
1880 Louis Pasteur demonstrated that injection of killed microbes provided
protection upon subsequent exposure to live counterpart
1888 Elie Metchnikoff demonstrated that certain blood cells ingest foreign
material
1894 Jules Bordet discovered complement
1897 Robert Kaus discovered precipitins
1901 Emil von Behring had the distinction of being awarded as the first
immunology-related Nobel Prize for his works on serum therapy
1984 Discovery of the T cell receptor gene
1987 Susumu Tonegawa was awarded the Nobel Prize for his 1978 discovery of
the genetic principles underlying the generation of antibodies with
different specificities

PROPERTIES OF NATURAL AND ACQUIRED IMMUNE SYSTEMS

 Natural immunity is the ability of the individual to resist infection by means of
normally present body functions
 Acquired immunity, in contrast, is a type of resistance that is characterized by
specificity for each individual pathogen and the ability to remember a prior
exposure, which result in an increase response upon repeated exposure

NATURAL /INNATE ACQUIRED /ADAPTIVE/ SPECIFIC

1. Present at birth 1. Not present at birth (acquired)
2. Standardized response 2. Diverse response for each antigen
3. Lacks memory (adaptive)
3. Recalls previous antigen/with memory

NATURAL ADAPTIVE
FIRST LINE OF DEFENSE SECOND LINE OF THIRD LINE OF
DEFENSE DEFENSE
1. Skin and mucous 1. Phagocytosis 1. Specialized
membrane 2. Inflammation and fever lymphocytes
2. Secretions - contain 3. Natural antimicrobial a. Tells
lysozyme that attacks substances b. B cells - plasma cells
bacterial cell wall a. Complement - ab 2. Antibodies
3. Acidity of the git and independent
vagina pathways
4. Cilia lining the b. Properdin
respiratory tract c. Interferon
d. Tumor necrosis
factor
e. Betalysin

COMPONENTS OF THE NATURAL IMMUNE SYSTEM

CELLULAR 1. Phagocytic cells ex. neutrophils, monocytes/macrophages
2. Basophils, mast cells
3. Eosinophils
4. Natural killer cells / Null lymphocyte
HUMORAL 1. Complement – ab independent pathways
2. Interferon
3. Lysozymes
4. 4. Natural antimicrobial substances

COMPONENTS OF THE ADAPTIVE SYSTEM

CELLULAR 1. T cells
2. B cells – plasma cells
HUMORAL 1. Lymphokines
2. Antibodies

PHAGOCYTOSIS
 The engulfment of cells or particulate matter by leukocytes, or other cells.

1. INITIATION
→ Phagocytosis is initiated as a result of tissue damage, either trauma or as a
result of microorganism multiplication
→ Activated phagocyte has increased surface receptors that allow for adherence
a. CR3
b. Laminin receptor
c. Leucyl-formyl-methionyl-phenylalanine receptor

2. CHEMOTAXIS
→ Process by which cells tend to move in a certain direction under the stimulation
of a chemical substance
a. Positive chemotaxis – towards the stimulus
b. Negative chemotaxis – away from the stimulus
→ Chemotactic substances: complement components ex. C5a
→ Without the influence of these chemotactic substances, cell motion is RANDOM

3. ENGULFMENT
→ By active amoeboid motion
→ Final structure is known as phagosome or phagocytic vacuole
→ Opsonins (antibodies and complement components) interact with the surfaces
of bacteria, rendering them acceptable to the phagocyte

4. DIGESTION
→ Minute cell particle that contain hydrolytic enzymes and peroxidase approach
the phagosome, fuse with it, rupture, and discharge contents to it
→ Cells become degranulated as foreign materials are digested
5. EXCRETION

DESTRUCTION OF THE ANTIGEN

1. Formation of the phagolysosome leading to the release of lysosomal contents
including defensins, lactoferrin and lysozyme.
2. Nitric oxide produced by activated macrophages, which is toxic to microorganisms.
3. Activation of the NADPH oxidase, which is present in the phagosome membrane,
leading to the production of reactive oxygen intermediates, superoxide anion,
hydrogen peroxide and hydroxyl radicals, which are cytotoxic for microorganisms.

INFLAMMATION
 Tissue reaction to injury (redness, tenderness, pain, swelling) to injury by physical
or chemical agents, including microorganisms.

CARDINAL SIGNS
1. Redness – rubor
2. Heat – calor
3. Tumor – swelling
4. Dolor – pain
5. Functio laesa – loss of function

STAGES
1. VASCULAR RESPONSE - injured cells release histamine: promotes
VASODILATION
a. Increased blood flow to the injured area (hyperemia)
b. Increased capillary permeability - plasma leakage to tissues

2. CELLULAR RESPONSE
a. Neutrophils - first to migrate at site of cell injury, short-lived (acute
inflammation)
b. Monocytes/macrophages - second to migrate at site of cell injury, long lived
(chronic inflammation)

Characteristics of Acute-Phase Reactants

PROTEIN RESPONS NORMAL INCREAS FUNCTION
E TIME CONCENTRATI E
(hr) ON (mg/dL)
C-reactive protein 6-10 0.5 1000x Opsonization,
complement
activation
Serum amyloid A 24 3.0 1000x Removal of
cholesterol
Alpha1-antitrypsin 24 200–400 2-5x Protease inhibitor
Fibrinogen 24 110–400 2-5x Clot formation
Haptoglobin 24 40–200 2-10x Binds hemoglobin
Ceruloplasmin 48–72 20–40 2x Binds copper and
oxidizes iron
Complement C3 48–72 60–140 2x Opsonization, lysis
Mannose-binding ? 0.15–1.0 ? Complement
protein activation

3. RESOLUTION AND REPAIR – initiated by fibroblast proliferation, as a result:

a. Affected area may be totally repaired
b. Injury may lead to the formation of an abscess with some loss of function
 c. Granuloma may be formed, typical of delayed hypersensitivity or cell-mediated
immunity

INTERFERON (IFN)
→ Family of glycoproteins produced by all animal cells that exert a VIRUS-
NONSPECIFIC but HOST-SPECIFIC antiviral activity.

TYPE 1 IFN: ALPHA AND BETA

→ Non-immune, produced primarily in the initial innate response to viral infection
IFN - ALPHA Produced by virus-induced leukocyte culture
Major producers: null lymphocytes
Also known as the LEUKOCYTE INTERFERON
IFN - BETA Produced by dsRNA fibroblast cells
Major producers: fibroblasts and epithelial cells
Also known as the
FIBROBLAST/EPITHELIAL/FIBROEPITHELIAL INTERFERON

TYPE 2 IFN: GAMMA

→ Immune, primarily produced as a component of the specific immune response to
viral and other pathogens
IFN - GAMMA Produced by immunologically-stimulated lymphocytes
Major producers: T cells
Also known as the IMMUNE INTERFERON

TUMOR NECROSIS FACTOR (TNF)

→ TNF – ALPHA: produced by macrophages, also known as CACHECTIN
→ TNF – BETA: produced by CD4+ and CD8+ cells, also Known as
LYMPHOTOXIN

COMPLEMENT
→ Series of proteins that are normally present in serum whose overall function is
the mediation of inflammation

PROPERDIN
→ Serum protein that exerts bactericidal and viricidal effects in the presence of C3
and MAGNESIUM

BETALYSIN
→ Heat-stable cationic substance with bactericidal activity found in the serum of
many animal species including humans

ACQUIRED/ADAPTIVE/SPECIFIC IMMUNITY

TYPE MODE OF ACQUISITION

ACTIVE Natural Infection (Ag)
Artificial Vaccination (Ag)
a. Live organisms (smallpox)
b. Attenuated or weakened organisms (BCG vaccine
for MTB - attenuated M. bovis) BACILLUS OF
CALMETTE AND GUERIN
c. Dead organisms (cholera, typhoid)
d. Toxoids (tetanus) - toxin teals) W chemical
e. Modified virus (poliovirus)
PASSIVE Natural Transfer in vivo (IgG) Ab
 Colostrum (IgA) Ab
Artificial Immune Serum (Igs) Abs

LYMPHOID ORGANS
1. PRIMARY LYMPHOID ORGANS
a. Thymus
b. Bone marrow
2. SECONDARY LYMPHOID ORGANS
a. Spleen
b. Lymph nodes
c. MALT, Peyer's patches, appendix
CELLS INVOLVED IN SPECIFIC IMMUNITY
T CELLS B CELLS
Develop in the thymus Develop in the bone marrow
End products of activation are cytokines End product of activation is antibody
Antigens include CD2, CD3, CD4, CD8 Antigens include CD19, CD20, CD21,
CD40, MHC class Il
Identified by rosette formation with Identified by surface immunoglobulins
sRBCs
Located in paracortical region of lymph Located in cortical region of lymph nodes
nodes

T LYMPHOCYTES
 Represent approximately 80% of the circulating lymphocytes in the peripheral
blood
 Most circulating T cells express three of the following CD markers:
 CD2: sheep red blood cell receptor, which is the classical T-cell surface marker
 CD3: part of T-cell antigen-receptor complex
 CD4: receptor for MHC class II molecule
 CD8: receptor for MHC class I molecule

Helper-inducer T-cells: CD4+ cells

Suppressor-cytotoxic T-cells: CD8+ cells
Normal ratio CD4+:CD8+ cells 2:1
In HIV, ratio of CD4+:CD8+ cells 0.5:1 or 1:2

T CELL DEVELOPMENT

1. Double-  Lack CD4 and CD8 markers, which are important to their later
negative function
thymocytes  Actively proliferate in the outer cortex under the influence of
interleukin-7
 Rearrangement of the genes that code for the antigen
receptor known as TCR begins at this stage CD3, the complex
that serves as the main part of the T-cell antigen receptor
2. Double-  Express both CD4 and CD8 antigens
positive
thymocytes
3. Mature T  Survivors of selection exhibit only one type of marker, either
cells CD4 or CD8, and they migrate to the medulla
 Not certain why one marker is down-regulated, but it may
depend on which MHC protein the cell interacts with, and
exposure to certain cytokines
 T cells recognize antigen along with MHC class Il protein, while
 CD8+ T cells interact with antigen and MHC class I proteins
 T cells bearing the CD4 receptor are termed helper, or inducer,
cells, while the CD8-positive population consists of cytotoxic T
cells
 Approximately two-thirds of peripheral T cells express CD4
antigen, while the remaining one-third express CD8 antigen
 These mature T cells are released from the thymus and seed
peripheral lymphoid organs
 Resting T cells have a life span of up to several years in these
peripheral organs
 T helper cells consist of two subsets, termed Th1 and Th2 cells
a. Th1 cells produce interferon gamma (IFN-Y) and tumor
necrosis factor-beta (TNF-B), which protect cells against
intracellular pathogens
b. Th2 cells produce a variety of interleukins, including IL-4, IL-
5, IL-10, and IL-13; the essential role of the Th2 cells is to
help B cells produce antibody against extracellular
pathogens
4. Activated T  Activated T lymphocytes express receptors for IL-2, just as
cells activated B cells do
5. Sensitized T  T lymphoblasts differentiate into functionally active small
cells lymphocytes that produce cytokines. Activities of specific
cytokines include assisting B cells in commencing antibody
production, killing tumor and other target cells, rejecting
grafts, stimulating hematopoiesis in the bone marrow, and
initiating delayed hypersensitivity allergic reactions
 Immune response is known as cell-mediated immunity

In addition to effector cells, T memory cells are also generated. They have a higher
affinity for antigen than unstimulated T cells, and they may represent precursors in
the later stage of the primary response. In a similar manner to memory B cells, they
are able to proliferate sooner than naive T cells. They also express a broader array
of cytokines and appear to persist for years.

B LYMPHOCYTES
• Bone-marrow derived lymphocytes
• Precursor cells in antibody production

1. Pro-B  Have distinctive markers that include surface antigens CD19,

cells CD45R, CD43, CD24, and c-Kit
 C-Kit on the pro-B cell interacts with a cell surface molecule called
stem cell factor, which is found on stromal cells
 Intracellular proteins found at this stage are terminal
deoxyribonucleotide transferase (TdT) and recombination-
activating genes RAG-1 and RAG-2, which code for enzymes
involved in gene rearrangement
 Rearrangement of genes on chromosome 14, which code for the
heavy-chain part of the antibody molecule
2. Pre-B  First heavy chains synthesized are the µ chains, which belong to
cells IgM
 The µ chains accumulate in the cytoplasm
 Pre-B cells may also express µ chains on the cell surface,
accompanied by an unusual light chain molecule called a
surrogate light chain
  Surrogate light chains consist of two short polypeptide chains that
are noncovalently associated with each other; they are not
immunoglobulin proteins but are thought to be essential for
regulating B-cell development
3. Immature  Distinguished by the appearance of complete IgM molecules on
B cells the cell surface; this indicates that rearrangement of the genetic
sequence coding for light chains on either chromosome 2 or 22
has taken place by this time
 Completion of light chain rearrangement commits a cell to
produce an antibody molecule with specificity for a particular
antigen or group of related antigens
 Variable regions, which occur on both the light and heavy chains,
determine this specificity
 Other surface proteins that appear on the immature B cell include
CD21, CD 40, and MHC class Il molecules
 CD21 acts as a receptor for a breakdown product of the
complement component C3, known as C3d; this enhances the
likelihood of contact between B cells and antigen, because
antigen frequently becomes coated with complement fragments
during the immune response
 CD40 and MHC class II are important for interaction of B cells with
T cells
4. Mature B  In addition to IgM, all mature B cells exhibit IgD
cells  Unless contact with antigen occurs, the life span of a mature B
cell is typically only a few days
 If, however, a B cell is stimulated by antigen, it undergoes
transformation to a blast stage, which eventually forms memory
cells and antibody-secreting plasma cells. This process is known
as the antigen-dependent phase of B-cell development. These B
cells have a half-life of more than 6 weeks.
5. Activated  Antigen-dependent activation of B cells takes place in the primary
B cells follicles of peripheral lymphoid tissue
 Exhibit identifying markers that include CD25, which is found on
both activated T and B cells and acts as a receptor for IL-2
 Transform into blasts that will give rise to both plasma cells and
so-called memory cell
6. Plasma  Characterized by the presence of abundant cytoplasmic
cells immunoglobulin and little to no surface immunoglobulin
 Represent the most fully differentiated lymphocyte, and its main
function is antibody production
 Not normally found in the blood but are located in germinal
centers in the peripheral lymphoid organs
 Nondividing, and after several days of antibody production, they
die without further proliferation.

SURFACE MARKERS ON T AND B CELLS

ANTIGE CELL TYPE FUNCTION
N
CD2 Thymocytes, T cells, NK Involved in T cell activation
cells
CD3 Thymocytes, T cells Associated with T cell antigen receptor; role
in TCR transduction
CD4 Helper T cells, monocytes Co-receptor for MHC class II; receptor for HIV
and macrophages
 CD5 Mature T cells, Positive or negative modulation of T and B
thymocytes, subset of B cell receptor signaling
cells (B1)
CD8 Thymocyte subsets, Co-receptor for MHC class I
cytotoxic T cells
CD10 B and T cell precursors, Protease; marker for pre-B CALLA
bone marrow stromal cells
CD16 Macrophages, NK cells, Low affinity Fc receptor, mediates
neutrophils phagocytosis and ADCC
CD19 B cells, follicular dendritic Part of B cell co-receptor, signal transduction
cells molecule that regulates B cell development
and activation
CD21 B cells, follicular dendritic Receptor for complement component C3d;
cells, subset of immature part of B cell co-receptor with CD19
thymocytes
CD23 B cells, monocytes, Regulation of IgE synthesis; triggers release
follicular dendritic cells of GM-CSF from monocytes
CD25 Activated T, B cells, Receptor for IL-2
monocytes
CD44 Most leukocytes Adhesion molecule mediating homing to
peripheral lymphoid organs
CD45R Different forms on all Essential in T and B cell antigen-stimulated
hematopoietic cells activation
CD56 NK cells, subsets of T cells Not known
CD94 NK cells, subsets of T cells Subunit of NKG2-A complex involved in
inhibition of NK cell cytotoxicity

LABORATORY IDENTIFICATION OF LYMPHOCYTES

1. CELL FLOW CYTOMETRY
→ Automated system for identifying cells based on scattering of light as cells flow
in single file through a laser beam
→ Forward LS: CELL SIZE
→ Side LS: CELL GRANULARITY

2. FLUORESCENCE MICROSCOPY
→ Both techniques rely on the use of labeled monoclonal antibodies against
specific surface antigens

3. OTHER METHODS: ROSETTE TEST

MAJOR HISTOCOMPATIBILITY COMPLEX

→ Genes that control expression of a large group of proteins
→ Regulate the immune response and play a role in graft rejection
→ Genes coding for the MHC molecules in humans are found in the short arm of
CHROMOSOME 6

CLASS I CLASS II CLASS III

Genetic loci HLA – A, B, C HLA – DP, DQ, DR C2, C4
Chain structure Alpha chain Alpha and beta Factor B
& Beta2- chain Tumor necrosis
microglobulin factor
Cell distribution All nucleated cells B cells and APCs (TNF)
Presents Ag To: CD8+ or Tc To: CD4+ or Th
cells cells
IMMUNOGEN
→ Substance that is capable of inducing an immune response

ANTIGEN
→ Substance with the ability to combine with an antibody

EPITOPE: The key portion of the immunogen against which the immune response is
directed; also known as the DETERMINANT SITE.

FACTORS AFFECTING IMMUNOGENECITY

1. FOREIGNNESS
 Autoantigen - from the same individual
 Alloantigen - from different individuals, but of the same species
 Heteroantigen- from different species
 Heterophile antigens- antigens from unrelated plants and animals, cross-
react with antibody of another

GRAFT
o Autograft - from the same individual
o Isograft/syngraft- from different but identical individuals
(IDENTICALTWINS)
o Allograft- from different individuals, but of the same species
o Heterograft - from different species
NOTE: TRANSPLANTATION IMMUNOLOGY
Examples of the Immunogenicity of Different Transplant Tissues
Most BONE MARROW
Immunogenic Skin
Islets of Langerhans Heart
Kidney
Liver
Bone
Xenogeneic valve replacements
Least CORNEA (PRIVILEGE SITE, AVASCULAR)
Immunogenic

CATEGORIES AND CHARACTERISTICS OF GRAFT REJECTION

TYPE TIME OF TISSUE PREDOMINANT CAUSE
DAMAGE MECHANISM
Hyperacut W/in minutes Humoral Preformed cytotoxic Abs to
e donor Ags
Accelerat 2-5 days Cell-Mediated Previous sensitization to
ed donor Ags
Acute 7-21 days Cell-Mediated Dev’t of allogeneic reaction
to donor Ags
Chronic >3 months Cell-Mediated Disturbance of host/graft
tolerance

2. SIZE
→ POTENT ANTIGEN: MW > 10, 000 DALTONS
→ Albumin: MW 40, 000 daltons, good immunogen
→ Hemocyanin: MW 1M daltons, excellent immunogen

3. CHEMICAL COMPOSITION AND COMPLEXITY

 → PROTEINS – MOST IMMUNOGENIC
→ Polysaccharides – second
→ Lipids, nucleic acid – least

4. ROUTE, DOSAGE AND TIMING

→ Generally, intravenous and intraperitoneal routes are effective; the intradermal
route offers stronger stimulus than the subcutaneous or intramuscular route
(although there are exceptions to this).
→ Dose response may be partially dependent on the nature of immunogen
processing. Generally, the smaller the dose, the less likely a response

5. ADJUVANTS
→ Substances added to vaccines and less immunogenic substances TO ENHANCE
IMMUNE RESPONSE
a. Complete Freund's Adjuvant (CFA): stimulates T cells
b. Lipopolysaccharides (LPS): stimulate B cells
c. Alum adjuvant: stimulates phagocytic cells

ANTIBODIES (IMMUNOGLOBULINS)
→ Substances produced in response to antigenic stimulation that is capable of
specific interaction with provoking immunogen

GENERAL FUNCTIONS OF IMMUNOGLOBULINS

1. Neutralize toxic substances
2. Facilitate phagocytosis and kill microbes
3. Combine with antigens on cellular surfaces and thereby cause the destruction of
these cells either extravascularly (outside of the blood vessels within the
mononuclear-phagocyte system) or intravascularly (within the blood vessels
through the action of complement)

THEORIES OF ANTIBODY DIVERSITY

1. Ehrlich's Side-Chain Theory

→ Paul Ehrlich formulated the side-chain theory in the early 1900s. Ehrlich
postulated that certain cells had specific surface receptors for antigen that were
present before contact with antigen occurred. Once antigen was introduced, it
would select the cell with the proper receptors, combination would take place,
and then receptors would break off and enter the circulation as antibody
molecules. New receptors would form in place of those broken off and enter the
circulation as antibody molecules

2. Template Theory
→ Felix Haurowitz put forth a second major theory, the instructive or template
theory in the early 1930s. According to this theory, antibody-producing cells are
capable of synthesizing a generalized type of antibody, and when contact with
an antigen occurs, the antigen serves as a mold or template and alters protein
synthesis so that antibody with a specific fit is made. This now-specific antibody
enters the circulation, while the antigen remains behind to direct further
synthesis

3. Clonal Selection
→ Niels Jerne and Macfarlane Burnet independently supported the idea of clonal
selection process for antibody formation. The key premise is that individual
 lymphocytes are genetically preprogrammed to produce one type of
immunoglobulin, and that a specific antigen finds or selects those particular
cells capable of responding to it, causing it to proliferate.

PROPERTIES OF IMMUNOGLOBULINS

IgG IgM IgA IgD IgE

Structure Monomer Pentamer Serum – Monomer Monomer
Monomer – monomer
B cells Sec – dimer
Percent of 70-75 10 10-15 <1 0.002
total
immunoglobu
lin
MW (Daltons) 150,000 900,000 160,000 180,000 190,000
Sedimentatio 7S 19 S 7S 7S 8S
n
coefficient
Serum half- 23 6 5 1-3[ 2-3
life (days)
C' fixation YES YES ALT NO NO
Crosses YES NO NO NO NO
placenta

TYPES OF IMMUNOGLOBULIN

IMMUNOGLOBULIN G

The major immunoglobulin in normal serum is IgG. It diffuses more readily than
other immunoglobulins into the extravascular spaces and neutralizes toxins or binds
to microorganisms in extravascular spaces. IgG can cross the placenta. In addition,
when IgG complexes are formed, complement can be activated. IgG accounts for
70% to 75% of the total Ig pool. It is a 7S molecule, with an MW of approximately
150,000 Daltons. One of the subclasses, IgG3, is slightly larger (170,000 Da) than
the other subclasses.

→ Predominant immunoglobulin in humans

→ Longest half-life of any immunoglobulin class
Subclasses: differ in the number and arrangement of disulfide bonds
IgG1 67% (2)
lgG2 22% (4)
lgG3 7% (15)
IgG4 4% (2)

MAJOR FUNCTIONS OF IgG:

1. Providing immunity for the newborn
 Cross placenta: except IgG2 (Henry, Stevens)
 Most efficient: IgG1
2. Fixation of the complement
 Classical pathway except IgG4
 Best subclass IgG3
  IgM > IgG3 > IgG1 > IgG2
3. Opsonization
4. Neutralization of toxins and viruses
5. Participation in agglutination and precipitation reactions

IMMUNOGLOBULIN M
It accounts for about 10% of the Ig pool and is largely confined to the intravascular
pool because of its large size. This antibody is produced early in an immune
response and is largely confined to the blood. IgM is effective in agglutination and
cytolytic reactions. In humans, IgM is found in smaller concentrations than IgG or
IgA. The molecule has five individual heavy chains, with an MW of 65,000
Daltons; the whole molecule has an MW of 900,000 Daltons and sedimentation
coefficient of 19s.
→ Known as the macroglobulin
→ Most primitive
→ First to appear in phylogeny and the last to leave in senescence
→ First to appear after a primary antigenic stimulus
→ Antibody most often formed in response to stimulus by gram negative bacteria

FUNCTIONS OF IgM:
1. Complement fixation: BEST/PENTAMERIC
→ Most efficient in triggering the classical complement pathway because a single
molecule can initiate the reaction due to its multiple binding sites
2. Agglutination
3. Opsonization
4. Neutralization of toxins
 Wasserman antibodies, heterophil antibodies, RF, cold agglutinins and
allohemagglutinins characteristically occur as IgM

IMMUNOGLOBULIN A
It represents 15% to 20% of the total circulatory Ig pool. It is the predominant
immunoglobulin in secretions such as tears, saliva, colostrum, milk, and intestinal
fluids. IgA is synthesized largely by plasma cells located on body surfaces. If
produced by cells in the intestinal wall, IgA may pass directly into the intestinal
lumen or diffuse into the blood circulation. As IgA is transported through intestinal
epithelial cells or hepatocytes, it binds to a glycoprotein called the secretory
component. The secretory piece protects IgA from digestion by gastrointestinal
proteolytic enzymes. It forms a complex molecule termed secretory IgA, which is
critical in protecting body surfaces against invading microorganisms because of its
presence in seromucous secretions (e.g., tears, saliva, nasal fluids, colostrum).
→ Subclasses IgA1 and IgA2
→ Primarily IgA2 is found as a dimer in secretions
→ Secretory component - prevents enzymatic degradation of IgA, produced by
epithelial cells near IgA-producing plasma cells

IMMUNOGLOBULIN D
It is found in very low concentrations in plasma, accounting for less than 1% of the
total lg pool. IgD is extremely susceptible to proteolysis and is primarily a cell
membrane Ig found on the surface of B lymphocytes in association with IgM.
→ Function IMMUNOREGULATION
→ Primarily a cell membrane immunoglobulin found on the surface of B
lymphocytes in association with IgM
→ Postulated to be an anti-idiotypic antibody (antibody to antibody) and as such
may be involved in a feedback mechanism to switch off B-cells.
IMMUNOGLOBULIN E
A trace plasma protein found in the blood plasma of unparasitized individuals (MW,
188,000 Da). IgE is crucial because it mediates some types of hypersensitivity
(allergic) reactions, allergies, and anaphylaxis and is generally responsible for an
individual's immunity to invading parasites. The IgE molecule is unique in that it
binds strongly to a receptor on mast cells and basophils and, together with antigen,
mediates the release of histamines and heparin from these cells
→ Least abundant immunoglobulin in the serum
→ HEAT-LABILE antibody, originally called REAGINIC ANTIBODY
→ Binds strongly to a receptor on mast cells and basophils and together with
antigen, mediates the release of histamine and heparin from these cells
→ Mediates some types of hypersensitivity (allergic) reactions, allergies, and
anaphylaxis and is generally responsible for an individual's immunity to
invading parasites

MONOCLONAL ANTIBODY (Georges Kohler and Cesar Milstein)

A very specific antibody derived from a single antibody-producing cell that has been
cloned or duplicated. Initially used for in vitro diagnostic testing. Recently however,
there has been an emphasis on the use of monoclonal antibodies as therapeutic
agents
A mouse is immunized, and spleen cells are removed. These cells are fused with
non- secreting myeloma cells and then plated in a restrictive medium. Only the
hybridoma cells will grow in this medium, where they synthesize and secrete a
monoclonal immunoglobulin specific for a single determinant of an antigen.

COMPLEMENT
→ A humoral mechanism of nonspecific immune response consisting of 14 distinct
serum proteins (nine components) that proceed in a cascading sequence of
activation, resulting in cell lysis.
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 - form C3 convertase
C3 Key intermediate in all pathways
C5 Initiates membrane attack complex
C6 Binds to C5b in MAC
C7 Binds to C5C6 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
MBL PATHWAY
MBL Binds to mannose
MASP-1 Helps cleave C4 and C2
MASP-2 Cleaves C4 and C2

The complement system proteins are named with a capital C followed by a number.
A small letter after the number indicates that the protein is a smaller protein
resulting from the cleavage of a larger precursor by a protease. Several complement
proteins are cleaved during activation of the complement system; the fragments are
designated with lower case suffixes, such as C3a and C3b. Usually, the larger
fragment is designated as "b" and the smaller fragment as "a." The exception is the
designation of the C2 fragments; the larger fragment is designated C2a and the
smaller fragment is C2b.

PLASMA COMPLEMENT REGULATION

C1 Inhibitor (C1INH) Dissociates C1r and C1s from C1q
Factor I Cleaves C3b and C4b
Factor H Cofactor with I to inactivate C3b; prevents binding of B
to C3b
C4-binding protein Acts as a cofactor with I to inactivate C4b
S protein (vitronectin) Prevents attachment of the C5b67 complex to cell
membranes

DEFICIENCIES OF COMPLEMENT COMPONENTS

DEFICIENT COMPONENT ASSOCIATED DISEASE
C1 (q, r or s) Lupuslike syndrome; recurrent infections
C2 Lupuslike syndrome; recurrent infections;
atherosclerosis
С3 Severe recurrent infections; glomerulonephritis
C4 Lupuslike syndrome
C5-C8 Neisseria infections
C9 No known disease association
C1 INH Hereditary angioedema
DAF Paroxysmal nocturnal hemoglobinuria
MIRL Paroxysmal nocturnal hemoglobinuria
Factor H or factor I Recurrent pyogenic infections
MBL Pneumococcal diseases, sepsis, Neisseria infections
Properdin Neisseria infections
MASP-2 Pneumococcal diseases

HYPERSENSITIVITY
→ Heightened state of immune responsiveness

COMPARISON OF HYPERSENSITIVITY REACTIONS

Type I Type II Type III Type IV
ANAPHYLACTI СУТОТОХІС IMMUNE DELAYED
C COMPLEX CELL-
MEDIATED
Immune IgE IgG, IgM IgG, IgM T cells
mediator
C’ No Yes Yes No
involvement
(classical)
Effector cells BASOPHILS, RBCs, WBCs, HOST TISSUE MACROPHAGES,
MAST CELLS PLTs CELLS TCELLS
Immune RELEASE OF CYTOLYSIS DEPOSITS OF RELEASE OF
mechanism MEDIATORS DUETO AB AG-AB CYTOKINES
MAST CELLS AND C’ COMPLEXES
AND
 BASOPHILS
Examples ANAPHYLAXIS TRANSFUSION SERUM CONTACT
HAY FEVER REACTIONS, SICKNESS DERMATITIS
FOOD AIHA, HDN ARTHUS TUBERCULIN
ALLERGIES REACTION TEST
ASTHMA SLE PNEUMONITIS

In the skin, histamine is responsible for local erythema or redness and wheal and
flare formation

TUMOR-ASSOCIATED ANTIGENS AND THEIR AREAS OF CLINICAL USE AND THE

ASSOCIATED TUMORS
ANTIGEN USES ASSOCIATED TUMOR/S
Bence Jones Protein 1 Multiple myeloma
AFP 1,2,3 Nonseminomatous testicular cancer, primary
hepatoma
hCG 1,2,3,4 Nonseminomatous testicular cancer,
choriocarcinoma
Calcitonin 1,2,3,4 Familial medullary thyroid carcinoma
PSA 1,2,3,4 Prostate cancer
CA- 125 1,2,3,4 Ovarian adenocarcinoma
Cytokeratins 2 Sarcomata, hematopoietic origin
HMB-45 2 Malignant melanoma
Neuron-specific enolase 2 Small cell cancers of the lung, endocrine
tumors
S-100 2 Neuroendrocrine tumors, melanoma
Beta-2 microglobulin 3 Lymphoma
Lactate dehydrogenase 3 Lymphoma
CEA 2,3,4,5 Tumors of the gastrointestinal tract
Estrogen receptor 3 Breast adenocarcinoma
CD45 2 Hematopoietic malignancies
IL-2 receptor (CD25) 4 T cell leukemia
Monoclonal Ig 1,2,3,4 Clonality indicates B cell malignancy
CA-19.9 4 Colonic and pancreatic adenocarcinoma
CA-15.3 4 Breast adenocarcinoma
Prostate specific 5 Prostatic adenocarcinoma
membrane antigen
NRLU-10 (Ab name) 5 Small cell lung cancer
B-1 5 Lymphoma
CD20 6 Lymphoma
MAGE 6 Melanoma
1= Screening 4= Monitoring
2= Pathologic diagnosis 5= Localization of metastasis
3= Staging 6= Therapy

TUMOR MARKERS (from Dean Rodriquez Clinical Chemistry Review

Handbook,2012)
TUMOR MARKERS ASSOCIATED CANCERS
AFP Hepatic and testicular cancers
ALP (placental-ALP) Lung cancer
Amylase Pancreatic cancer
BRCA-1 Breast or ovarian cancer
CA-125 Ovarian cancer (treatment and recurrence)
CA-15.3 Breast cancer (treatment and recurrence)
CA-19.9 Gastric, pancreatic and colorectal cancers
CA-50 Gastric and pancreatic cancers (treatment and recurrence)
CA-27.29 Breast cancer (treatment and recurrence)
Calcitonin Medullary thyroid cancer
Cathepsin-D Breast cancer
CEA Colorectal, stomach, breast, lung cancer (treatment and
recurrence)
CK-1 Small cell lung cancer, prostate cancer
Estrogen receptor (ER) Breast cancer
GGT Hepatoma
HER-2/neu Breast cancer (efficiency of trastuzumab or herceptin
therapy)
Nuclear matrix protein Urinary bladder cancer
(NMP)

AUTOIMMUNE DISEASES
→ A condition in which damage to body organs results from the presence of
autoantibodies or autoreactive cells

General Features of Some Autoimmune Disorders That Have Systemic, Multi-

Organ, or Multi-Tissue Involvement
Autoimmune Disease Ratio HLA
F/M
RHEUMATOID ARTHRITIS 3:1 HLA-DR4
 Mixture of organ specific with systemic symptoms
 Inflamed, swollen, and painful joints
 RF- IgM to the Fc region of IgG
 Peak age of onset is 40-50 years of age
SYSTEMIC LUPUS ERYTHEMATOSUS 10:1 HLA-DR2
 Systemic and multi-organ HLA-DR3
 Immune complexes are formed in serum
 Immune complexes are trapped in basement
membrane of glomeruli, skin/endothelium, synovial
joints, kidney
 ANA, anti-ENA and anti-dsDNA may be present
 Anti-phospholipid antibodies (lupus anticoagulant) are
present
MYASTHENIA GRAVIS 4:1 Complex
 Tissue sp (nerve and muscle)-with systemic effects Age
 Neuromuscular transmission disorder dependent
 Antibodies are inhibitory and block acetylcholine
binding
 Some evidence of prior infection with poliovirus
SCLERODERMA 3:1 HLA-DR3
• Primarily in skin/ectodermal tissues, but can cause multi- (weak)
organ pathology
• Skin fibroblasts reproduce faster and produce more
collagen
SJOGREN' S SYNDROME 10:1 HLA-DR3
 Clinically presents as dry eyes and mouth
 It can affect any of the body's glands that produce
sweat, saliva or oil
IMMUNOLOGICAL AND PATHOLOGICAL FEATURES OF SOME AUTOIMMUNE
DISORDERS
DISORDER PATHOLOGY ANTIBODIES
Type 1 Diabetes Selective destruction of the Anti-glutamic acid
insulin producing β cells of decarboxylase (GAD)
the islets of Langerhans in antibodies (B islet cell
the pancreas antigen)
Anti-insulin antibodies
Rheumatoid Arthritis Chronic systemic RF: IgM or IgG antibodies
inflammatory disorder, in specific for the Fc region of
which joint cartilage, IgG
ligaments and tendons are
destroyed
Systemic Lupus Immune complexes are ANA
Erythematosus formed and these lodge in Anti-ENA
the basement membrane Anti-dsDNA
of kidney, skin and joints Anti-phospholipid (lupus
anticoagulant)
Graves' Disease Unregulated secretion of Anti-TSH receptor antibodies
T3 and T4 due to
stimulation of TSH receptor
by antibody
Hashimoto's Destruction of the thyroid Anti-thyroid peroxidase
Thyroiditis gland antibodies
(anti-microsomal antibodies)
Anti-thyroglobulin antibodies
Pernicious Anemia Destruction of the parietal Anti-parietal cell antibodies
cells of the stomach Anti-intrinsic factor antibodies
mucosa leading to intrinsic
factor deficiency

IMMUNODEFICIENCIES
DEFICIENCIES OF THE B CELL SYSTEM
1. Transient Hypogammaglobulinemia of Infancy
→ Infants experience low levels of immunoglobulins at approximately 5 to 6
months of age, but in some babies the low levels persist for a longer time
→ Cell-mediated immunity is normal, and there may be normal levels of lgA and
IgM. IgG tends to be the most affected.
→ Immunoglobulin levels usually normalize spontaneously, usually by 9 to 15
months of age

2. X-linked Bruton' s Agammaglobulinemia

→ X-chromosome-linked, affects males almost exclusively
→ Patients lack circulating B cells and exhibit deficiency or lack of
immunoglobulins of all classes

3. IgA Deficiency
→ Selective IgA deficiency is the most common congenital immunodeficiency
→ Most patients are asymptomatic. Those with symptoms usually have infections
of the respiratory and gastrointestinal tract and an increased tendency to
autoimmune diseases such as SLE, RA and thyroiditis. Allergic disorders and
malignancy are also more common.

4. Common Variable Immunodeficiency (CVI)

 → Characterized by hypogammaglobulinemia that leads to recurrent bacterial
infections, particularly sinusitis and pneumonia
→ There is usually a deficiency of both IgA and IgG, but selective IgG deficiency
may occur

5. Isolated IgG Subclass Deficiency

→ Isolated deficiency of any one of the four IgG subclass

DEFICIENCIES OF CELLULAR IMMUNITY

1. DiGeorge Anomaly
→ Developmental abnormality of the third and fourth pharyngeal pouches that
affects thymic development
→ Quantitative defect in thymocytes; not enough mature T cells are made but
those that are present are functionally normal

2. Purine Nucleoside Phosphorylase (PNP) Deficiency

→ PNP deficiency affects an enzyme involved in the metabolism of purines
→ It produces moderate to severe defect in cell-mediated immunity with normal or
mildly impaired humoral immunity

COMBINED DEFICIENCIES OF CELLULAR AND HUMORAL IMMUNITY

1. Severe Combined Immunodeficiency (SCID)
→ Most serious of the congenital immunodeficiencies
→ Group of related diseases that all affect T and B cell function

2. Wiskott-Aldrich Syndrome (WAS)

→ Triad of immunodeficiency, eczema and thrombocytopenia

3. Ataxia-Telangiectasia (AT)
→ Characterized by cerebellar ataxia and telangiectasias, especially on the
earlobes and conjunctiva
→ Levels of IgG2, IgA and IgE are often low or absent; in addition the number of
circulating I cells is often decreased

DEFECTS OF NEUTROPHIL FUNCTION

1. Chronic Granulomatous Disease (CGD)
→ Most common and best characterized of the neutrophil abnormalities
→ Genetic defect in any of the several components of the NADPH oxidase system
can result in the CGD phenotype by making the neutrophils incapable of
generating an oxidative burst

2. Leukocyte Adhesion Deficiency (LAD)

→ In LAD, a protein (CD18) that is a component of adhesion receptors in
neutrophils or monocytes (with CD11b or CD11c) and on T cells (with CD11a) is
defective
→ Defect leads to abnormal adhesion, motility, aggregation, chemotaxis and
endocytosis of the affected leukocytes