Antigens, haptens and

immunogens
Definitions
Immunogen

Antigen (Ag)
Hapten

Epitope or Antigenic Determinant

Antibody (Ab)
 Definitions:

Anantigen is any substance that react with T or B

lymphocytes

simple to macromolecules e.g. carbohydrates,

phospholiplids, nucleic acids and proteins.
 Immunogens are substances that generate
immune response
 Haptens are small molecules which could
never induce an immune response unless
coupled to a carrier molecule (foreign protein)

e.g. dinitrophenyl, aminobenzene sulphonate,

arsonate

Haptein-carrier molecule, unlike free

haptein, can acts as an immunogen.
 Antibodyrecognizes antigen through
complementariness of shapes on paratope and
epitope.

The forces are unspecific between two unrelated

proteins:-
Electrostatic
Hydrogen bonding
Hydrophobic
Van der waals
 The affinity or strength of binding of antigen
and antibody depends on closeness of fitting.

Itrefers to strength of association between

an individual epitope and a paratope, high
affinity antibodies bind strongly to antigen.
 Avidity is the strength of binding two molecules or
cells to one another at multiple sites.
It is determined by

heterogeneity of antibodies in serum

heterogeneity of antigenic determinants

bonus effect = k avid = k1 x k2

Avidity measure of the functional affinity of an

antiserum for the whole antigen.
 The specificity of antigen recognition by an
antibody is not absolute

Cross-reactivity is due to antiserum reacting with

a partially related antigen.

Monoclonal antibodies are directed towards a

single epitope less cross reactivity.
 Any antibody binds to a specific portion of a macromolecule called
epitope or determinant

The part of the hypervariable region on the antibody which contacts

the antigen is termed as the paratope.

If the antigen is a liner peptide or carbohydrate the combining site

can accommodate 5-6 amino acid residues or hexose units.

With globular proteins as many as 16 a. acids may contact the

antibody.

Antigens and antibodies interact by spartial complementarity not by

covalent bonding.

Two types of epitopes:-

Discontinous or assembled
Continous or sequential
 Antigenic determinant is a cluster of
epitopes on the surface of an antigen.
Antigen has several determinants each
structurally different from each other.
A monoclonal antibody reacts with only one
determinant on the same antigen.
The overall configuration of a haptein is
more important than its chemical structure
in determining the strength of binding to an
antibody.
Factors Influencing
Immunogenicity
Contribution of the Immunogen
Foreignness

Size

Chemical Composition
Primary Structure Sequence determinants
Secondary Structure
Tertiary Structure Conformational
determinants
Quarternary Structure
Factors Influencing
Immunogenicity
Contribution of the Immunogen
Foreigness

Size

Chemical Composition
Physical Form
Particulate > Soluble
Denatured > Native
Factors Influencing
Immunogenicity
Contribution of the Immunogen
Foreigness

Size

Chemical Composition
Physical Form

Degradability
Ag processing by Ag Presenting Cells (APC)
Factors Influencing
Immunogenicity
Contribution of the Biological System
Genetics
Species
Individual
Responders vs Non-responders
Age
Factors Influencing
Immunogenicity
Method of Administration
Dose

Route
Subcutaneous > Intravenous > Intragastric
Adjuvant
Substances that enhance an immune response to
an Ag
Chemical Nature of
Immunogens
Proteins

Polysaccharides

Nucleic Acids
Lipids
Some glycolipids and phosopholipids can be
immunogenic for T cells and illicit a cell mediated
immune response
Types of Antigens
T-independent

Polysaccharides
Properties
Polymeric structure
Polyclonal B cell
activation
Yes -Type 1 (TI-1)

No - Type 2 (TI-2)

Resistance to
degradation
Examples
Pneumococcal polysaccharide, lipopolysaccharide
Flagella
Types of Antigens
T-dependent
Proteins

Structure
Examples
Microbial proteins
Non-self or
Altered-self
proteins
Hapten-carrier conjugates
Definition

Structure
Haptenic determinants
native
determinants
haptenic
determinants
Native determinants
Antigenic Determinants
Recognized by B cells and Ab

Composition
Proteins, polysaccharides, nucleic acids
Sequence (linear) determinants
Conformational determinants
Size
4-8 residues
Antigenic Determinants
Recognized by B cells and Ab

Composition
Size
Number
Limited Fe
(immunodominant
epitopes)
Located on the
external surfaces
of the Ag
Antigenic Determinants
Recognized by T cells
Composition
Proteins (some lipids)
Sequence determinants
Processed
MHC presentation (lipid presentation by MHC-like
CD1)
Size
8 -15 residues
Number
Limited to those that can bind to MHC
 What Does The B Cell Immunoglobulin
(Ig) Receptor Recognize?

1. Proteins (conformational determinants,

denatured or proteolyzed
determinants)
2. Nucleic acids
3. Polysaccharides
4. Some lipids
5. Small chemicals (haptens)
What Does the T Cell
Receptor (TCR) Recognize?
1. Only fragments of proteins (peptides)
associated with MHC molecules on
surface of cells
Helper T cells (Th) recognize peptide
associated with MHC class II molecules
Cytotoxic T cells (Tc) recognize peptide
associated with MHC class I molecules
Antigen Processing and
Presentation
Fragmentation of protein into peptides
Association of peptide with an MHC
molecule
Transport to cell surface for expression
Different cellular pathways for
association of peptide with MHC class I
and class II molecules
 Class I MHC Pathway
Peptide is presented
by MHC-I to CD8
cytotoxic T cell Viral protein is made
Plasma membrane on cytoplasmic
ribosomes

Globular viral
protein - intact
Peptide passes
with MHC from Golgi Proteasome
body to surface rER degrades
Peptide associates protein to
with MHC-I complex peptides
Peptide transporter
protein moves
peptide into ER
MHC class I alpha
Golgi body Peptide with MHC
and beta proteins
goes to Golgi body
are made on the rER
 Class II MHC Pathway
Peptide MHC-II
complex is presented CD4 helper T cell Globular
to CD4 helper T cell protein

Endosome fuses with Endosome

Endocytosis
plasma membrane Fusion of endosome
and exocytic vesicle
Immunodominant Lysosome
peptide binds
to class II MHC
Exocytic vesicle fuses
with endosome Protein is processed to
Golgi releasing Ii from dimer peptides in endosome
body or lysosome

Class II MHC
Synthesis
Ii 3 chains: , and Ii
Endoplasmic reticulum
Points Concerning Antigen
Processing and Presentation
1. Location of pathogen
viruses in cytosol, MHC class I pathway,
Tc response

extracellular bacteria, MHC class II

pathway, Th2 response, Ab formation
intracellular bacteria, MHC class II
pathway, Th1 response
Superantigens
Proteins produced by pathogens
Not processed by antigen presenting
cells
Intact protein binds to variable region of
chain on TCR of T cells and to MHC
class II on antigen presenting cells
(APC)
Large numbers of activated T cells
release cytokines having pathological
effects
Conventional Antigen
Superantigen
Antigen presenting cell

CHO 2 2 CHO 2 2
MHC Class II
CHO 1 1 CHO CHO 1 1 CHO
Super
Antigen
antigen
CHO CHO CHO CHO
V V V V
T cell receptor
CHO CHO CHO CHO
C C C C

T lymphocyte
 Summary
antigen, haptein, immunogen
epitope
paratope
Ag-Ab reactions spatial complementarily, not
covalent bonds

Only weak forces are involved

Affinity
avidity bonus effect of multivalency
cross - reactivity