Antigens and Immunoglobulins

Suzan Matar (PhD Medical Microbiology and Immunology)

Department of Clinical Laboratory Sciences
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Introduction

Antibodies are circulating proteins that are

produced in vertebrates in response to
exposure to foreign structures known as
antigens.

An antigen is any substance that may be

specifically bound by an antibody molecule or
T cell receptor.

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 An antigen is any agent capable of binding
specifically to B cell receptor (BCR) or T-cell
receptor (TCR).

The compound that can evoke an immune

response can be called either Antigen or
Immunogen.

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 Typical immunogens include pathogenic
microorganisms (e.g. bacteria, viruses, and
parasites), foreign tissue graft, proteins in pollen,
grass, or food.

A microorganism may contain several

immunogens.

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Epitope (antigenic determinant): is the specific
antigenic site of the antigen molecule.

An epitope is a peptide of
just 4 or 5 amino acids, or a
monosaccharide in size. An
antigen molecule may have
one or more epitopes

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 Haptens are usually small molecules, e.g. penicillin.

Haptens are antigenic but not immunogenic.

Hapten becomes immunogen when combines with a carrier

protein.

Hapten-carrier protein complex triggers production of anti-

hapten antibody with which the hapten reacts specifically.

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Hapten: a low molecular weight molecule that can be
made immunogenic by conjugation to a suitable carrier

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 CHEMICAL NATURE OF IMMUNOGENS

A. Proteins
The vast majority of immunogens are proteins. These
may be pure proteins or they may be glycoproteins or
lipoproteins. In general, proteins are usually very good
immunogens. They contain multiple epitopes.

B. Polysaccharides
- Pure polysaccharides and lipopolysaccharides are weak
immunogens.
- Glycoproteins
- Teichoic acids and ABO.
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 C. Nucleic Acids (need carrier)
- Nucleic acids are usually poorly immunogenic. However,
they may become immunogenic when single stranded or
when complexed with proteins.

D. Lipids (need carrier)

- In general lipids are non-immunogenic, although they
may be haptens.
- Could be glycolipids

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Cross reactivity of Antigen
Results from structural
homology with certain parts
of two different antigens.
Sharing enough epitops to
allow the immune response.

Homologous e.g.: toxin vs toxoid

Heterologous e.g. : heterophil antigens (share same 3-D

characteristics.

1. Blood Group A pneumococcal capsular polysaccharide

type XIV
2. Blood Group B certain E. coli Antigen.
3. Smallpox cowpox vaccine.
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 Antigenicity and specificity
Common antigen and cross-reaction

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Primary and secondary immune responses

The first exposure to an immunogen is

referred to primary immunization which
generates a primary response.

A second exposure to the same immunogen

results in a secondary phase.

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Antibody Structure and Function

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Antibodies are synthesized only by cells of the B
lymphocyte lineage and exist in two forms:

1. membrane bound antibodies on the surface of B

lymphocytes function as antigen receptors,

1. and secreted antibodies neutralize toxins,

prevent the entry and spread of pathogens, and
eliminate microbes. (plasma cells )

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 When blood or plasma forms a clot, antibodies
remain in the residual fluid, which is called
serum.

Any serum sample that contains detectable

antibody molecules that bind to a particular
antigen is commonly called an antiserum.

The study of antibodies and their reactions with

antigens is therefore classically called serology.

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Immunoglobulins
change during infection
and in certain diseases
such as Myeloma.

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 The most two important features of immunoglobulins:
- Specificity
- Biological activity

Neutralize toxins and viruses,

Opsonize microbes so they are more easily
phagocytosed,
Activate complement,
Prevent the attachment of microbes to mucosal
surfaces.

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 Antibody structure
Antibodies Are Made Up Of:
2 Light Chains (identical) ~25 KDa
2 Heavy Chains (identical) ~50 KDa
Each Light Chain Bound To Heavy Chain By Disulfide (H-
L)
Heavy Chain Bound to Heavy Chain (H-H)
Globular domains bind by hydrophobic interactions
First 100 a/a Of amino terminal vary of Both H and L
Chain Are Variable, referred To As VL andVH
CDR (Complementarity Determining Regions) Are What
Bind Ag = hypervariable regions
Remaining Regions Are Very Similar Within Same Class
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Isotypes/classes (Heavy chains, Constant
region)
IgM
IgG
IgD
IgE
IgA
- Subclasses (differ in number of disulfide bonds)
- IgG1,2,3,4
- IgA1,2

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Antibody structure

CDR (Complementarity Determining

Regions) Are What Bind Ag =
hypervariable regions

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Binding affinity

Two antibodies with different CDR to have

specificity for the same epitopes but different
affinities due to differences in the number
and type of binding forces .

Binding affinity: measure of the strength of

the binding.

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Cross-reactivity : antibody of a given specificity can
exhibit binding with two different epitopes (Ag1 and Ag2)
(Redundancy)

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I. IgG
u Structure: Monomer
u Percentage serum antibodies: 80%
u Location: Blood, lymph, CSF and peritoneal fluid
u Half-life in serum: 23 days
u Complement Fixation: Yes
u Placental Transfer: Yes

u Known Functions:
- Enhances phagocytosis,
- neutralizes toxins and viruses,
- protects fetus and newborn. 35
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3. IgG can do Opsinization

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 Antivirus Ab

Ab

FcRg Apoptosis
Programmed
NK
CTL Cell Death
Perforins = (pore forming protein)+
Granzymes
4. Antibody Dependent Cell-mediated Cytotoxicity
ADCC

5. Activation of complement
6. Immunity/ neutralization of toxins
7. Neutralization of viruses
8. Immobilization of bacteria
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II. IgM
u Structure: Pentamer
u Percentage serum antibodies: 5-10%
u Location: Blood, lymph, B cell surface (monomer)
u Half-life in serum: 5 days
u Complement Fixation: Yes
u Placental Transfer: No

u Known Functions:
- First antibodies produced during an infection. First
Ig made by fetus and B cells.
- Effective against microbes and agglutinating
antigens.
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Biological Functions of IgM

IgM is BCR

Primary response

Agglutination / Problems in
blood transfusion.

Activation of complement
(classical pathway)

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III. IgA
u Structure: Dimer
u Percentage serum antibodies: 10-15%
u Location: Secretions (tears, saliva, intestine, milk)
as sIgA, blood and lymph.
u Half-life in serum: 6 days
u Complement Fixation: No
u Placental Transfer: No
u Known Functions: Localized protection of mucosal
surfaces. Provides immunity to infant digestive
tract.
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 Transcytosis of dimeric IgA
Biological Functions of IgA across epithelia

Most produced antibody, yet

mostly in secretions.

Role in mucosal infections.

(agglutination).

Bactericidal activity especially

against gram-negative in the
presence of lysozyme.

Antiviral activity.

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IV. IgD
u Structure: Monomer
u Percentage serum antibodies: 0.2%
u Location: B-cell surface, blood, and lymph. Not
secreted by plasma cells. It is membrane Ig.
u Half-life in serum: 3 days
u Complement Fixation: No
u Placental Transfer: No
u Known Functions: In serum function is unknown. On B
cell surface, initiate immune response.
u IgD may be in silencing autoreactive B cells.
u In mature B cells it serves as an antigen- binding
surface immunoglobulin with coexpressed IgM
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V. IgE (reaginic antibody)
u Structure: Monomer
u Percentage serum antibodies: 0.002%
u Location: Bound to mast cells and basophils throughout
body. Blood.
u Half-life in serum: 2 days
u Complement Fixation: No
u Placental Transfer: No
u Known Functions: Allergic reactions (histamine, heparin,
leukotrienes). Possibly lysis of worms.

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IgE's main function is
Immunity to parasites
such as parasitic worms like
Schistosoma mansoni,
Trichinella spiralis, and
Fasciola hepatica.

IgE may also be important

during immune defense
against certain protozoan
parasites such as
Plasmodium falciparum.

IgE also plays an essential

role in type I
hypersensitivity, such as
allergic asthma, allergic
rhinitis, food allergy, and
dermatitis.

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Monoclonal Antibodies

A tumor of plasma cells (myeloma or

plasmacytoma).

The resultant fused cells that grow out are

called hybridomas; each hybridoma makes
only one Ig, derived from one B cell from the
immunized animal.

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Monoclonal Ab applications

Identification of phenotypic markers unique

to particular cell types. (CD of lymphocytes)
Immunodiagnosis.
Tumor identification.
Therapy. (complications serum sickness)
Functional analysis of cell surface and
secreted molecules.

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ISOTYPES, ALLOTYPES, & IDIOTYPES

Because immunoglobulins are proteins, they

are antigenic, and that property allows them
to be subdivided into isotypes, allotypes, and
idiotypes.

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(1) Isotypes are defined by antigenic (amino acid)
differences in their constant regions.

(2) Allotypes, on the other hand, are additional

antigenic features of immunoglobulins that vary
among individuals.

(3) Idiotypes are the antigenic determinants formed

by the specific amino acids in the hypervariable
region.

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Ig variants

Differences in
the constant
region of H
and L chains=
Gm, Km, Am

Iso similar
Allo different
Idio- private, own

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Antibody Functions

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