11/26/2014

Antibody: Isotype; Allotype & Idiotype

Herd immunity
Monoclonal Antibodies
Autoimmunity: Proposed mechs. of
pathogenesis
Transplantation Immunology
Immunodeficiency Diseases

Antibody: Isotype; Allotype & Idiotype

Dr Debasis Biswas

Isotypic determinants
Determinants located in the constant region
Collectively define:
Heavy- chain class & subclass;
Light chain type & subtype
within a species
All members of a species carry the
same constant region genes

How to obtain antibodies to:

Isotypic determinants

Allotypic determinants

Injecting ab from one

species to another species.
Example: Anti- human IgG
raised in sheep

Injecting ab from one

member of a species to
another member.
Example: Blood transfusion;
Pregnant mother raising
ab to paternal allotypes
present in fetal Ig

Allotypic determinants
Within the same species,
some genes have multiple alternative forms
Certain epitopes are present in some
members of the species.. Allotypic determinant
Aggregate of all the allotypic determinants
displayed by an antibody determines its Allotype

Idiotypic determinants
Epitopes present in the
Variable region of Light & Heavy chains
Each individual epitope present in the
variable region .. Idiotypic determinant
Aggregate of all the idiotypic determinants
displayed by an antibody determines its Idiotype

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A Susceptible population
(Pre- vaccination era)

An Immunized population
(100% coverage)

Herd immunity
An Immunized population
(<100% coverage)
Infected host

Susceptible host

Immunized host

Herd Immunity
Immunizing agents transmitted from the vaccine recipients
to the non- recipients
Non- vaccinated agents derive the benefit of immunization
by sharing the same environment as the vaccinees
Circulation of vaccine strains minimize the chances of
contact between pathogenic wild strains and the
susceptible hosts
Examples: Polio
Measles

Monoclonal Antibodies

Polyclonal antibody response

Epitopes

Antigen molecule
with multiple epitopes

Polyclonal
antiserum
Experimental animal
(mouse, g pig, rabbit, sheep, etc)

Every ag . Multiple epitopes

Every epitope Proliferation & Differentiation of a
clone of B cells . Secretion of antibodies specific
for the epitope
Mixture of ab: Polyclonal ab response ......
Facilitates the localization, phagocytosis, complementmediated lysis of a pathogen in vivo

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Hybridoma Technology

Monoclonal Antibodies: Diagnostic Uses

Epitopes

Detection of:
Pathogens, through identification of specific
antigens (Hepatitis B surface Ag, HBsAg, for HBV; p24 for

Monoclonal
Antibodies
Ag injected into
mouse

HIV)
Hybridomas

Spleen cells
isolated

Pregnancy, through identification of hormonal

markers (HCG)
Tumors, through identification of tumor antigens
(Prostate Specific Ag, PSA)

Hybridize

Specific MHC ags for HLA typing

Measuring of:
Drug levels in blood (Digoxin)
Hormonal levels in blood (T3, T4, TSH)

Fused with
Myeloma cells

Monoclonal Antibodies: Diagnostic Uses

Monoclonal Antibodies: Therapeutic Use

Monoclonal Imaging Technique

Radiolabelled monoclonal antibodies
Targeted towards specific tumor antigens
Early detection of 1ry or metastatic tumors in
patients
Example: I131 labelled monoclonal abs to Breast
Cancer ags, injected into bloodstream, can detect
metastasis to regional LNs

Monoclonal Antibodies: Prophylactic Use

Humanized Murine Monoclonal Ab (Hypervariable
regions from mouse; rest of the molecule: human
ab) to Respiratory Syncitial Virus (RSV) protects
from severe respiratory disease in infants.

Autoimmunity: Proposed
mechanisms of Pathogenesis

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Overview
Failure of self- tolerance
Earlier theory: Failure to eliminate self- reactive
lymphocytes in the Thymus or Bone marrow (Central
Tolerance)
Presently: Failure of the phenomenon of clonal anergy
or clonal suppression that regulates self- reactive
lymphocytes (Peripheral Tolerance)
Mode of tissue destruction: Antibody- mediated (e.g.
Autoimmune Hemolytic Anemia; Hashimotos
Thyroiditis) or Cell mediated (Rheumatoid Arthritis;
IDDM; Multiple Sclerosis)
Categories: Organ- specific (Thyroiditis; Goodpasture
Syndrome) or Systemic (SLE; RA)

II. Molecular mimicry

Resemblance in conformation or 1ry sequence
between a self- ag and a pathogen protein
Cross- reactivity of host abs or T cells with self ag s
Example: Post- rabies Encephalitis (cross- reactivity
between rabbit brain- cell ag s present in rabies
vaccines and host brain cell ag s)
Post- Streptococcal Rheumatic fever (cross- reactivity
between Streptococcal ag s and host cardiac ags)

I. Release of sequestered antigens

Thymus: Clonal deletion of self- reactive immature T
cells.. Self Tolerance
Ag s sequestered from circuluation would not induce
self tolerance
Exposure of mature T cells to such sequestered ags
may induce their activation
Example: Autoab formation resulting from.
Release of lens proteins following injury, or
Release of heart muscle ag s following MI

III. Inappropriate expression of MHC

Class II molecules
Trauma or viral infection in an organ
Localized inflammation in the organ .
Increased secretion of IFN .
Induces inappropriate expression of MHC Class II
molecules in the organ
Sensitization of Th cells to peptides derived from that
organ
Example: Pancreatic Beta cells in IDDM patients;
Thyroid acinar cells in Graves disease patients express
MHC Class II molecules

IV. Polyclonal B cell activation

molecules
Non- specific activation of numerous B cell clones
following infection with agents, like Cytomegalovirus,
Epstein Barr virus, Gram- negative bacteria
Activation of self- reactive B cell clones .
Production of autoantibodies
Example: Infectious mononucleosis (EB virus) leads to
production of autoantibodies to T cells, B cells,
Rheumatoid factor, Antinuclear antibodies
HIV. Opportunistic infection with EBV or CMV.
Autoantibody production against RBCs, platelets

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Overview

Transplantation Immunology

Overview
MHC genes= Major, but not the only, determinants of
histocompatibility.
Matching of MHC Class II ag is more important
determinant of graft survival than MHC Class I.
Minor Histocompatibility genes also involved.
Mismatch at no. of minor loci can also lead to graft
rejection
Graft rejection mediated by both CD4 and CD8 T cells
MHC proteins: Recognized directly by recipent Th &
CTLs
Minor HC proteins: Recognized only when they are
presented by self MHC molecules

Mechanism of graft rejection ..

Effector phase

Autograft= Self tissue. Example: Transferring healthy

skin to a burnt area. Healthy blood vessels replacing
blocked coronary arteries
Isograft= Tissue transfer between genetically identical
individuals. Example: Homozygotic twins
Allograft= Tissue transfer between genetically
different individuals of same species
Xenograft= Tissue transfer between members of
different species

Mechanism of graft rejection ..

Sensitization phase
CD4 and CD8 T cells of recipient recognize disparate
Major and Minor Histocompatibility ags (alloantigens)
within the graft tissue
Alloantigens presented by:
APCs (Dendritic Cells)
Endothelial cells
Stimulation of Recipient Th cells:
Th cell proliferation
Induction of effector mechanisms

Hyperacute Rejection...
Pre-formed antibodies to graft ags

Pre-existing abs
carried to the graft

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Primary Immunodeficiency Secondary Immunodeficiency

Genetic or developmental
Acquired loss of immune
defect in immune system
function secondary to various
factors like HIV/AIDS or
Present at birth; though may
immunosuppressive t/t
manifest later

Immunodeficiency Diseases

Manifestations
Phagocytic Deficiency: Bacterial infections
B cell deficiency: Bacterial infections
(Most commonly, infections with encapsulated bacteria like
Staphylococcus, Streptococcus & Pneumococcus, as antibody is
critical for opsonization and clearance of these bacteria).

T cell deficiency: May affect both

Orange box: Phagocytic deficiency
Green box: Humoral deficiency
Red box: CMI deficiency
Purple box: Combined deficiency

Humoral immunity: Bacterial infections

Cell- mediated immunity: Viral, protozoan & fungal infections
(Most commonly, infections with intracellular pathogens like
Candida albicans, Pneumocystis carinii, M. tuberculosis).

Combined deficiency: Most critical

Chronic Granulomatous Disease (CGD)

Inheritance: X linked or Autosomal Recessive
Defect in the oxidative pathway, which generates H2O2
and other reactive products like HOCl in phagocytes
Inability of phagocytes to kill phagocytosed bacteria
Susceptible to bacterial & fungal infections.
Exception: Pneumococcus (generates H2O2 )
Excessive inflammation: Gingivitis; Swollen LNs;
Chronic granulomas (subcutaneous lumps)
Also: deficiency of Mes to function as APCs ed CMI
IFN: Possible therapy to restore function of granulocytes

Leukocyte Adhesion Deficiency

Integrin molecules:
Adhesion of WBCs to Vasc. Endoth. .. Recruitment to
inflammatory sites
Interaction between T & B cells
Three Integrins on monocytic cells
LFA-1
CD11a/CD18

Mac-1
CD11b/CD18

gp150/95
CD11c/CD18

Defect in CD18. Defective recruitment of leukocytes

Susceptibility to Gram+ve & Gram-ve bacteria; fungi;
viruses (due to defective T & B cell interaction)

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Severe Combined Immunodeficiency

(SCID)

Severe Combined Immunodeficiency

(SCID)

Various forms of defects

Affect T cells/ both T & B cells
Myeloid & Erythroid cells appear normal in no.
Thymus does not develop
Low nos. of circulating T cells; which cannot
proliferate in response to ags
Severe recurrent infections (usually fungal & viral. Bacterial

infections late: maternal ab)

Fatal early in life

Even Live attenuated vaccines can cause infection and
disease

Wiskott Aldrich Syndrome

ADA/ PNP Def:

Interferes with purine
Metab. & DNA synthesis

X- linked Agammaglobulinemia

X- linked
Involves a cytoskeletal glycoprotein, sialophorin, reqd
for assembly of actin filaments
Initially T & B cells are present in normal nos.
Severity increases with age
Ds manifests itself by defective responses to bacterial
polysaccarides and lower levels of IgM
Gradually: recurrent bacterial infections;
thrombocytopenia
lymphoid malignancy
eczema

X- linked Agammaglobulinemia

X- linked Hyper IgM

X- linked
Defect in signal transduction molecule, Brutons
Tyrosine kinase (Btk)
No peripheral B cells
Extremely low IgG levels & absence of other Ig classes
Recurrent bacterial infections , beginning at about 9
months of age

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X- linked Hyper IgM

X- linked recessive
Defect in CD40L molecule in T cells, which leads to failure
of co-stimulation of B cells through CD40-CD40L
interaction. Failure of:
Class switching to IgG, IgA, IgE classes
Memory B cell production
Formn of Germinal Centers during humoral response
Elevated levels of IgM with:
Severe recurrent infections
Autoantibodies to neutrophils, platelets, RBCs

Common Variable Immunodeficiency

Genetic defect, though manifests later in life

Underlying defect not known
B cells fail to differentiate into plasma cells
Low levels of most Ig clases
Recurrent bacterial infections
T/t: Administration of Ig

Selective IgA deficiency

Commonest type of selective Ig deficiency
IgA- secreting B cells fail to differentiate into plasma cells
IgG2 & IgG4 may also be deficient
May be asymptomatic
Manifest as recurrent RTI/ GUTI, reflecting the role of IgA
in mucosal defence
Other manifestations:
Intestinal malabsorption
Allergy
Autoimmune diseases