Antigen, Antibody and Antigen-Antibody Reactions
Antigen, Antibody and Antigen-Antibody Reactions
Antigen, Antibody and Antigen-Antibody Reactions
ANTIGEN-ANTIBODY REACTIONS
ANTIGENS
• Definition : An antigen is a high molecular weight protein or
polysaccharide which when introduced into an immunocom -
petent host, generates an immune response stimulating T
cells or B cells or both .
• It should satisfy 2 properties –
Immunogenicity : Ability of an antigen to induce immune
response in the body
Antigenicity : Ability of an antigen to combine specifically with
antibodies or T cell surface receptors .
• Epitope / antigenic determinant : It is the smallest unit of
antigenicity that specifically binds to the antibody ( paratope).
• Hapten : These are low molecular weight molecules that lacks
immunogenicity but retain antigenicity . They become
immunogenic when combined with a larger protein molecule
called carrier . They are of 2 types :
Complex haptens : Contain 2 or more epitopes .
Simple haptens : Usually contain only one epitope .
• Antigen-Host relationship :
Self / autoantigens : They belong to the host itself and due to
immunological tolerance they are not immunogenic . But if
they are biologically altered , they can become
immunogenic ,e.g. in autoimmune diseases .
Non-self or foreign antigens : They are immunogenic and are
of 3 types based on their phylogenetic distance to the host –
Alloantigens – species specific antigens .
Isoantigens – present only in subsets of a species e.g. blood
group antigens .
Heteroantigens – antigens belonging to 2 different species e.g.
Heterophile antigen , exists in unrelated species like – Weil-
Felix reaction, Paul-Bunnell test and cold agglutination test .
• Factors influencing immunogenicity :
1. Size and chemical nature of antigen
2. Susceptibility of antigen to tissue enzymes
3. Structural complexity and foreignness to the host
4. Genetic factor
5. Optimal dose or route of antigen administration
6. Repeated doses of antigen
7. Multiple antigens ( like the use of Adjuvants )
8. Effect of prior administration of antibody
• Adjuvants : Any substance that enhances the immunogenicity
of an antigen . These are used in vaccines to enhance the
immunogenicity of vaccine antigens .e.g. LPS of Bordetella
pertussis acts as an excellent adjuvant for diphtheria and
tetanus toxoids .
• Biological Classes of antigens :
1. T- dependent antigen : most of the normal antigens .
2. T-independent antigen : like bacterial capsule , flagella and
LPS .
3. Superantigens : like toxic shock syndrome toxin ( TSST-1),
Streptococcal pyrogenic exotoxin (SPE)- A and C ,etc.
• Superantigens :
The variable beta region of T cell receptor is the receptor for
superantigens
They can activate T cells directly without being processed by
antigen presenting cells . Nonspecific activation of T cells
leads to polyclonal B cell activation leading to
hypergammaglobulinemia .
Diseases associated with superantigens are : Toxic shock
syndrome ,food poisoning ,scalded skin syndrome , atopic
dermatitis ,kawasaki syndrome ,etc.
ANTIBODY
• IgG :
1. Constitutes 70-80% of total Ig in the body .
2. It can cross placenta so provides immunity to the fetus and
new born .
3. It can activate the classical pathway of complement system
4. Plays role in phagocytosis by binding to the Fc receptors on
phagocytes .
5. Raised IgG levels represents chronic or past infections
6. Plays major role in neutralization of toxins
7. IgG subclass ( except IgG3) mediates coagglutination reaction
by binding to protein-A of S.aureus .
• IgM :
1. It has highest molecular weight and maximum sedimentation
coefficient . It is pentameric ( 10 Fab regions , 10 valencies ).
2. Present only in intravascular compartment , not in body fluids or
secretions . Protection against blood invasion by harmful
microorganisms .
3. Raised in primary immune response ( in acute infection )
4. Most potent activator of classical complement pathway .
5. Acts as an opsonin so enhances phagocytosis
6. First antibody to be synthesized in fetal life ( 20 weeks) , so
provides immunity to the fetus .
7. Mediates agglutination .
• IgA :
1. Exists in 2 forms : serum IgA(monomeric ) and secretory
IgA( dimeric ). Constitutes 10-15% of total serum Ig .
2. Serum IgA plays role in antibody-dependent cell mediated
cytotoxicity (ADCC) and degranulation of immune cells .
3. Secretory IgA is predominant antibody found in body
secretions , mediates local or mucosal immunity .
4. Breast milk is rich in secretory IgA , provides good protection
to the immunologically immature infant’s gut .
• IgE :
1. Lowest serum concentration, shortest half life and minimum
daily production .
2. It is heat labile ( inactivated at 56 degree centigrade for 1 hr)
3. Has affinity for surface tissue cells of the same species
(homocytotropism )
4. Mainly extravascular in distribution
5. It is highly potent and mediates type 1 hypersensitivity
reactions , its response is seen in asthma , anaphylaxis ,etc.
6. It is elevated in helminthic infections .
• IgD :
1. It is found as membrane Ig on the surface of B cells and acts
as a B cell receptor along with IgM .
2. It has the highest carbohydrate content among all Ig .
• Antigenic determinants of immunoglobulins :
1. Isotypes : Vary from each other in amino acid sequences of
constant region of their heavy chains . E.g. 5 classes of Ig . It
is similar in all members of same species .
2. Idiotypes : The unique amino acid sequence present in
paratope region of one member of a species acts as antigenic
determinant to other members of the same species .
3. Allotypes : The antigenic determinants present in the isotype
genes in constant region of H and L chains , encoded by
multiple alleles called as allotypes .
Examples of abnormal immunoglobulins :
• CONVENTIONAL TECHNIQUES :
1. Precipitation reaction
2. Agglutination reaction
3. Complement fixation test
4. Neutralization test
• NEWER TECHNIQUES :
1. ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA)
2. Enzyme-linked fluorescent assay (ELFA) – Ag-Ab complex
detected by fluorometric methods .
3. Immunofluorescence assay (IFA)
4. Radioimmunoassay (RIA)
5. Chemiluminescence –linked immunoassay (CLIA)
6. Immunohistochemistry
7. Rapid tests –
Lateral flow assay ( immunochromatographic test )
Flow through assay
8. Western blot
9. Immunoassays using electron microscope
PRECIPITATION REACTION :