Sie sind auf Seite 1von 50

IMMUNE RESPONSE

Biochemistry
Lecture 5.5

Prof. Dal Vieelien


Department of Biochemistry
E-mail: daleveze@med.kmu.lt

OVERVIEW OF ADAPTIVE IMMUNITY


Uses two basic strategies
Humoral Immunity
- works to eliminate antigens that are extracellular
Cellular Immunity
- deals with antigens within host cell

Humoral Immunity

IMMUNOGLOBULINS Ig
(ANTIBODIES)

Glycoprotein molecules that are produced by plasma cells in


response to an immunogen and which function as antibodies
Receptors for antigen on B-cells
When secreted immunoglobulin (antibodies)
One B-cell makes only one kind of antibody
Immunoglobulins comprise approx. 20 % proteins of blood
plasma

Antigen recognition is mediated


by specific antigen receptors
Antigen - any structure which elicits an immune
response
Recognition is a central feature of the immune
response, required for detection and elimination of
dangerous organisms (pathogens)
This recognition is mediated by specific antigen
receptors which bind to structures (antigens) on, or
derived from, pathogens.
Antigen receptors can be secreted or cell-associated

Antigenic determinant (epitope)

Structure of Immunoglobulins
Disulfide bond

Carbohydrate

CL
VL
CH2

CH1
VH

Hinge Region

Variable & Constant Regions


VL & CL
VH & CH
Hinge Region

CH3

2 identical Light chains


or each about 220
amino acids
2 identical Heavy chains a, d, e, g, or m - each about
450 amino acids
2 + 2
Disulfide bonds:
Inter-chain
Intra-chain

Oligosaccharides

Domains (Domains are folded,


compact, protease resistant
structures)
VL & CL
VH & CH1 - CH3 (or CH4)

A typical antibody molecule


Fab

Carbohydrate

CHO

CHO

Fc

Fab - fragment
antigen binding
Fc - fragment
crystallisable
Fc effector
functions. Fc
interacts with
innate effector
mechanisms and
greatly amplifies
them

Immunoglobulins are
Bifunctional Proteins

Pepsin cleavage sites


Papain cleavage sites

- 1 x (Fab)2 & 1 x Fc
- 2 x Fab 1 x Fc

Antigens can bind in


pockets or grooves or on
extended surfaces in the
binding site of antibodies
by non-covalent forces

Why do antibodies need an Fc region?


The (Fab)2 fragment can:

Detect antigen

Precipitate antigen

Block the active sites of toxins or pathogen-associated


molecules

Block interactions between host and pathogen-associated


molecules

The Fc fragment can activate:

Inflammatory and effector functions associated with cells

Inflammatory and effector functions of complement

The trafficking of antigens into the antigen processing


pathways

Consequences of antibody-antigen
binding
A. Viral Inhibition: virus preventing it from attaching to cell
B. Neutralization: make toxins unable to bind to cells
C. Opsonization: antibodies bind to antigen and facilitate
attachment of phagocytic cells
D. and E. Agglutination and Precipitation: antibodies bind to
antigen and get them into clumps, then one big mouthful
for phagocyte
F. Phagocytosis: Fc portion of antibody encourages
phagocytosis
Complement Activation: binding of antigen to antibody can
trigger one pathway of complement cascade

The complement system


Interacting set of enzymes (proteases) that upon activation give
rise to cascade of reactions culminating in the destruction of
pathogens and infected cells

Classes of immunoglobulins
(depend on H chain)

ANTIBODY CLASSES AND SUBCLASSES


(ISOTYPES)
Classes: IgM, IgG, IgD, IgA, IgE
Subclasses - IgG1, IgG2, IgG3, IgG4

MONOMER

PENTAMER

MONOMER

MONOMER WHEN
ACTING AS B-CELL
RECEPTOR FOR ANTIGEN

DIMER

MONOMER

IgG
(Monomeric)
IgG1, IgG2 and IgG4

IgG3

Major serum Ig
Major Ig in extravascular spaces 70-80% of total serum
antibody
Half-life in serum 23 days
Placental transfer (IgG2)
Fixes complement (IgG4)
Binds to Fc receptors (IgG2, IgG4)
Phagocytes - opsonization

The pentameric IgM molecule


Extra domain (CH4)
J chain
First Ig made by fetus and B cells
5-10% of total serum antibody
Half-life in serum 5 days
Fixes complement
Agglutinating Ig
Binds to Fc receptors
B cell surface Ig

First line of defence

Dimeric IgA- protection of the mucosa

Serum monomer
Secretions (sIgA) - major secretory Ig (Mucosal or Local Immunity)
10-15 % of total serum antibody (if mucous membranes and body secretions
included, percentage is much higher)
Half-life in serum 6 days
Tears, saliva, gastric and pulmonary secretions
Dimer: J chain; Secretory component
Does not fix complement (unless aggregated)
Binds to Fc receptors on some cells

IgD (Monomer)

0,2% of total serum antibody


Half-life in serum 3 days
B cell surface Ig, blood, lymph
Does not bind complement, serum function
not known

Immunoglobulin StructureFunction Relationship


Secreted antibody
Neutralisation

Arming/recruiting effector cells


Complement fixation
Cell surface antigen receptor on B cells
Allows B cells to sense their antigenic
environment

Connects extracellular space with intracellular


signalling machinery

IgE (Monomer)

Extra domain CH4


Least common serum Ig
0,002% of total serum antibody
Half-life in serum 2 days
Binds to basophils and mast cells
Allergic reactions
Parasitic infections (Helminths)
Binds to Fc receptor on eosinophils
Does not fix complement

Diversity: Antibody diversity is


generated by:
Multiple genes encoding both VL and VH

Segmental joining of additional DNA segments to form the


mature VL and VH genes
Addition of nucleotide bases during the joining event
Random association of Heavy and Light chains (facilitated by
the VC, CH1 disulfide bond)
Somatic (body) mutation of mature heavy and light chain
genes after antigen stimulation generate higher affinity
antibodies

Variable domains in immunoglobulins are made


by combinatorial joining of gene segments

Immunoglobulins and Development:


Fetal synthesis of IgM and IgA begin during the 5th month.
Immature B-cells express IgM on surface. IgM is monomeric on
surface.
Mature B-cells express IgM and IgD on surface.

Plasma cells can secrete IgM, IgG, IgA, IgE (all but IgD)
Memory cells display IgG, IgA, IgE, alone or combined with
IgM. These are usually higher affinity than the orginal B-cell
clone because of affinity maturation via somatic cell mutation

The immunoglobulin superfamily of


proteins
The structural hallmark of
immunoglobulin structure is the
existence of globular domains with
-pleated sheet folding pattern.

Variety of other proteins


which exhibit amino acid
sequence homology with Ig
also contain Ig-like domains
and are considered as
members of the
immunoglobulin superfamily

The genes that encode these


proteins have evolved from a
common ancestor gene coding
for a single domain.

The immunoglobulin superfamily of proteins

B Cell Antigen Receptor (BCR)

Ig-

Ig-
Ig-

Ig-

B-cells and Antibody Response


B-cell receptor binds to antigen
One of two things happen:
B-cell needs confirmation by T-cell to begin
responding
B-cell does not need confirmation by T-cell to
begin responding

When B-cell does not need


confirmation from T-cell
B-cell receptors bind epitopes
B-cells respond by proliferating, producing
antibody and differentiating into memory Bcells

When B-cell needs confirmation from T-cell

CELLULAR IMMUNITY

Types of T cells (cellular immune


response)

Also called CD8 T cells


Once activated, induce apoptosis in
self cells infected with virus; destroy
cancerous host cells
Distinguish infected self cells, because
these cells present peptides on surface in
MHC class I molecule

Also called CD4 T-cells


Antigen presenting cells, present
antigen to T-helper cells in MHC
class II
If recognize antigen presented as
foreign, activate macrophages,
release cytokines that recruit other
cells of immune system, stimulate
NK cells, activate B cells

T Cell Receptor (TCR)


TcR molecule resembles Fab and is monovalent

The TCRs are


transmembrane,
insoluble proteins.
Approximately 105
TcR molecules are
present on the surface
of a T cell.
Consists of an and
chain, or a and
chain.

In human majority of T cells express the heterodimer;


the remaining T cells (small percentage) express the
heterodimer (no , or T cells exist).
Each chain has a variable (V) and a constant domain (C).

TCR Structure
The variable domains in both chains contain three
hypervariable regions which are equivalent to CDRs present in
Ab light and heavy chains.
Transmembrane domains of both chains contain positively
charged amino acid residues
The most TCRs interact with peptide antigens presented by
MHC molecules.
Certain T cells react with nonpeptide antigens
(carbohydrate and lipids)
In contrast to TCR, TCR exhibits limited diversity
The T cells react with antigen (e.g. phospholipid antigen of
Mycobacterium tuberculosis) that is neither processed or
presented in the context of a MHC molecule.

TCR chain receptor gene

rearrange similar to antibody genes

Comparison of TCR and BCR


(antibody)
BCR
Any structure
Ligand
Bind native ligand Yes
No
Ag processing
MHC restriction No
Somatic mutation Yes
Affinity for ligand Low to very high
No
Co-receptors

TCRa
peptide & lipid
No
Yes
Yes
No
Low
Yes

T cells recognize the antigen only when it is


presented by self-MHC molecule.
This phenomenon, called self-MHC
restriction, differentiates recognition of
antigen by T cells and B cells.
The MHC is highly polymorphic from individual
to individual

Single receptor
recognizes
an alteration in selfMHC molecules induced
because of association
with foreign antigen.

Class I molecules are expressed on the surfaces of virtually all


nucleated cells at varying densities
Class II molecules are more restricted to cells of the immune
system, primarily B lymphocytes and monocytes.
Class I molecules function at the effector phase of immunity by
presenting antigens to CD8+ T cells, which generally have cytotoxic
or suppressor function
Class II molecules present antigenic fragments to the CD4+ inducer
(or helper) T cells on antigen presenting cells such as macrophages
MHC class I molecules present peptides derived from cytosolic
proteins; the pathway is called the cytosolic or endogenous pathway
MHC class II molecules present peptides derived from extracellular
proteins; the pathway is called the endocytic or exogenous pathway.

Epitopes recognized by TCR does not


need to lie on the surface of the protein

Interplay between T-cells and B-cells