4 - Antigen Recognition - 3ed (Compatibility Mode)

ANTIGEN RECOGNITION IN THE
ADAPTIVE IMMUNE SYSTEM

Sitti Wahyuni, MD, PhD
Department of Parasitology,
Medical Faculty, Hasanuddin University
5/13/2014
S.Wahyuni/lecture/immunology/S1
DURING LECTURE/DISCUSSION
5/13/2014
The principal function of cellular receptors is to detect external

stimuli and trigger responses of the cells on which the receptors
are expressed.
Antigen receptors of lymphocytes must be able to bind to and
distinguish between many, often closely related, chemical
structures.
Antigen receptors on lymphocyte B and T are clonally distributed:
each clone having a particular specificity has a unique receptor
The total number (repertoire) of lymphocyte specificities is very
large
Antigen receptors transmit biochemical signals that are
fundamentally the same in all lymphocytes and are unrelated to
specificity
5/13/2014
3
Questions to be addressed
How do the antigen receptors of lymphocytes recognize
extremely diverse antigens and transmit quite conserved
activating signals to the cells?
How is the vast diversity of receptor structures generated in
lymphocytes?
5/13/2014
Antigen Receptors of Lymphocytes

Features of antigen receptors of B and T lymphocytes
The antigen receptors of B and T lymphocytes recognize
chemically different structures.
B lymphocyte antigen receptors (membrane-bound
antibodies) are able to recognize the shapes, or
conformations, of native macromolecules (proteins, lipids,
carbohydrates, and nucleic acids and simple small chemical
groups)
T cells see only peptides that are displayed on antigenpresenting cells (APCs) bound to MHC
5/13/2014
Antigen receptor molecules consist

The antigen-recognizing portions: variable (V)
regions
The conserved portions: constant (C) regions.
Within the V regions there are variability which
are called hypervariable regions, or
complementarity-determining regions (CDRs)
5/13/2014
Antigen receptors are noncovalently attached to other

invariant molecules whose function is to deliver signal to the
inside of the cell
The collection of antigen receptors and signaling molecules
in B lymphocytes: B cell receptor (BCR) complex,
in T lymphocytes: T cell receptor (TCR) complex.
When bind to antigen molecules, the receptors are pulled
together into an aggregate (cross-linking)
Cross linking will catalyze enzymes that are attached to the
cytoplasmic portions of the signaling proteins and leads the
phosphorylation of other proteins.
Phosphorylation triggers complex signaling cascades that
culminate in the transcriptional activation of many genes
5/13/2014
Properties of antibodies and T cell antigen receptors (TCRs).

5/13/2014
Antibodies
Antibody molecule
is composed of 4 polypeptide chains=
2 identical heavy (H) chains & 2 identical light (L) chains
Each chain containing variable region & constant region
The four chains are assembled to form a Y-shaped molecule.
Each light chain is attached to one heavy chain
2 heavy chains are attached to each other by disulfide bonds.
A light chain is made up of one V and one C domain
A heavy chain has one V and three or four C domains.
Each domain folds into a characteristic three-dimensional
shape, which is called the immunoglobulin (Ig) domain.
5/13/2014
The structure of antibodies: immunoglobulins G (IgG) and M (IgM).

5/13/2014
10
Variable region
Regio of heavy & light chain that recognize & bind
to antigen
In heavy chain (called VH) or of the light chain
(called VL)
Contains three hypervariable regions (CDRs), the
greatest variability is in CDR3, which is located at
the junction of the V and C regions
Constant region
Region that is not attached to
antigen
5/13/2014
11
Regions of antibody based on the properties

of proteolytic fragments of immunoglobulins.
Fab (fragment antigen binding)
Contains a whole light chain (with its single V and C
domains) attached to the V and first C domains of a
heavy chain
Required for antigen recognition
Fc (fragment cystalline)
Contain the remaining heavy chain
Hinge region
Between the Fab and Fc regions
A flexible portion called the hinge region & allows the 2
antigen-binding Fab regions of each antibody molecule
to move independently of each other
5/13/2014
12
Light chain
There are two types of light chains, called and ,
that differ in their C regions but do not differ in
function.
Each B cell expresses either or , but not both
Heavy chain
There are five types of heavy chains, called , , , ,
and , that also differ in their C regions
Antibodies that contain different heavy chains are
said to belong to different isotypes, or classes, and
are named according to their heavy chains (i.e., IgM,
IgD, IgG, IgE, and IgA)
5/13/2014
13
Features of the major isotypes (classes) of antibodies

5/13/2014
14
Antibodies are capable of binding a wide variety of

antigens, (macromolecules and small chemical)
because antigen-binding regions of antibody
molecules form flat surfaces capable of
accommodating many different shapes
Antibodies bind to antigens by reversible,
noncovalent interactions, including hydrogen bonds
and charge interactions.
The parts of antigens that are recognized by
antibodies are called epitopes, or determinants.
5/13/2014
15
Binding of an antigen by an antibody

5/13/2014
16
Affinity
The strength with which one antigen-binding surface of an
antibody binds to one epitope of an antigen
Is expressed as the dissociation constant (Kd) = is the molar
concentration of an antigen required to occupy half the
available antibody molecules in a solution; the lower the Kd,
the higher the affinity.
In a primary immune response the Kd between 10-6 to 10-9 M
In a secondary immune response, the Kd between 10-8 to 1011 M
Affinity maturation: the increase in antigen-binding strength
5/13/2014
17
Antigen-binding site
The site where antibody bind to epitope
Each IgG, IgD, and IgE antibody molecule has two
antigen-binding sites
IgA has four antigen-binding sites
IgM has 10 antigen-binding sites
The total strength of binding is much greater than the
affinity of a single antigen-antibody bond and is called
the avidity of the interaction.
Antibodies produced against one antigen may bind
other, structurally similar, antigens cross-reaction.
5/13/2014
18
Monoclonal antibodies
Antibody that has similar spesificity
Has far-reaching implications for clinical medicine and research
Methods
An animal is immunized with an antigen x
B cell will produce plasma cells specific for x antigen
Plasma cells is taken from animal spleen and fused with
myeloma cell line that had been treated with toxic drug to
inhibit its grow
Fused cells grow and are called hybridomas
From a population of hybridomas, it is possible to select and
clone the continuously growing cells that secrete the
antibody x
5/13/2014
19
5/13/2014
20
Most of antibody monoclonal are made by fusing cells

from immunized mice with mouse myelomas
These mouse monoclonal antibodies cannot be injected
repeatedly into human subjects it will be regarded by
immune system as foreign
The solution: the antigen-binding V regions is retained
& the rest is replaced with human Ig "humanized"
antibody
5/13/2014
21
B cell receptors for antigens

Antigen receptor of B cell
(B cell receptor-BCR)
Is the antibody M or B that located
at the membrane of B cells
Composed of
2 light chains and 2 heavy chain
Each chain containing one variable (V) region constant region
Only heavy chains are anchored in the plasma membrane
Produced in a secreted form as effector
Undergo class switching or affinity maturation during the life
of a B cell clone.
5/13/2014
22
T cell receptors for antigens

Antigen receptor of T cell (T cell receptor-TCR)
Displayed by MHC molecules
A membrane-bound heterodimer
Composed of an chain and a chain
Each chain containing one variable (V) region and one
constant (C) region The V region of each TCR chain has 3
hypervariable, or complementarity-determining (CDR) and the
CDR3 is the most variable among different TCRs.
Both TCR chains are anchored in the plasma membrane
TCRs are not produced in a secreted form
TCRs do not undergo class switching or affinity maturation
during the life of a T cell clone.
5/13/2014
23
4-5 The structure of the T cell antigen receptor (TCR)

5/13/2014
24
and the chain of the TCR participate in specific

recognition of MHC molecules and bound peptides
Each TCR recognizes 1-3 residues of peptide
Note: only a few peptides of even complex microbes,
called the immunodominant epitopes, are actually
recognized by the immune system.
This means that T cells can tell the difference
between complex microbes on the basis of very few
amino acid differences between the
immunodominant epitopes of the microbes.
5/13/2014
25
The recognition of a peptide-MHC complex

by a T cell antigen receptor
5/13/2014
26
TCR
A lymphocyte T in innate immunity
About 5% - 10% of total T cells in the body
Structurally similar to the TCR but have very
different specificities
May recognize a variety of protein and nonprotein
antigens, usually not displayed by classical MHC
molecules.
Abundant in epithelia.
5/13/2014
27
NK-T cells
Comprising less than 5% of all T cells
Express markers of natural killer (NK) cells and are
called NK-T cells.
NK-T cells express TCRs, but they recognize
lipid antigens displayed by nonpolymorphic class I
MHC-like molecules
The functions of NK-T cells also are not well
understood.
5/13/2014
28
Features of antigen recognition by immunoglobulins (Igs) and

T cell antigen
receptors (TCRs).
5/13/2014
29
Development of Immune Repertoires

How the enormous diversity of these B and T cels receptors is
produced?
There are many clones of lymphocytes with distinct
specificities, perhaps as many as 109, and these clones arise
before encounter with antigen.
There are not enough genes in the human genome for every
possible receptor to be encoded by a different gene.
The immune system has developed mechanisms for
generating extremely diverse repertoires of B and T
lymphocytes
The generation of diverse receptors is intimately linked to the
process of lymphocyte maturation.
5/13/2014
30
Maturation of lymphocytes
The maturation of lymphocytes from bone marrow stem cells
consists of three types of processes:
proliferation of immature cells
expression of antigen receptor genes
selection of lymphocytes that express useful antigen
receptors .
5/13/2014
31
Lymphocyte maturation
Involving random genetic recombination events
Proliferation of lymphocyte stimulated mainly by interleukin-7
(IL-7) produced by stromal cells in the bone marrow and the
thymus.
IL-7 maintains and expands the number of lymphocyte
progenitors
After antigen receptor proteins are expressed, these receptors
take over the function of delivering the signals for
proliferation
5/13/2014
32
Steps in the maturation of lymphocytes

5/13/2014
33
Antigen receptors are encoded by several gene

segments that are separate from one another in the
germline and recombine during lymphocyte
maturation.
Diversity is generated during this recombination
process mainly by varying the nucleotide sequences at
the site of recombination.
The expression of diverse antigen receptors is the
central event in lymphocyte maturation
5/13/2014
34
Maturing lymphocytes are selected at multiple steps

during their maturation to preserve the useful
specificities.
Selection is based on the expression of intact antigen
receptor components and what they recognize.
Positive selection: Only select lymphocytes that
express antigen receptor
Negative selection: Only select lymphocytes that
has low affinity to self antigens present in the
bone marrow and thymus
5/13/2014
35
Production of diverse antigen receptors

The expression of B and T lymphocyte antigen
receptors is initiated by somatic recombination of
gene segments that code for the variable regions of
the receptors, and diversity is generated during this
process.
Hematopoietic stem cells in the bone marrow as well
as early lymphoid progenitors contain Ig and TCR
genes in their inherited, or germline, configuration.
5/13/2014
36
Each loci of heavy & light chain of BCR and & chain of TCR
contain
Multiple variable region (V) genes
One or a few constant region (C) genes
Several small stretches of nucleotides, called joining (J)
Diversity (D) gene segments.
All antigen receptor gene loci contain V, J, and C genes
The Ig heavy chain and TCR loci also contain D gene segments
Gene recombination is combination V, J, and C segment genes
(with or without D) the segments are selected randomly
5/13/2014
37
5/13/2014
38
5/13/2014
Recombination andS.Wahyuni/lecture/immunology/S1
expression of immunoglobulin (Ig) genes
39
The somatic recombination of V and J, or V, D, and J, gene

segments is mediated by a group of enzymes collectively, VDJ
recombinase.
The lymphoid-specific component of the VDJ recombinase
(recombinase-activating gene (RAG)-1 and RAG-2 proteins)
recognizes DNA sequences that flank all antigen receptor V, D,
and J gene segments.
The recombinase brings the V, D, and J segments close together
and cleaves the DNA at specific sites.
The DNA breaks are then repaired by ligases, producing a fulllength recombined V-J or V-D-J gene without the intervening DNA
segments
The lymphoid-specific component of the VDJ recombinase is
expressed only in immature
B and T lymphocytes.
5/13/2014
40
Diversity of antigen receptors is produced by:

the use of different combinations of V, D, and J gene
segments in different clones of lymphocytes (called
combinatorial diversity)
by changes in nucleotide sequences introduced at the
junctions of V, D, and J gene segments (called junctional
diversity)
Combinatorial diversity is limited by the number of available V,
D, and J gene segments, but junctional diversity is almost
unlimited.
This junctional diversity is produced by three types of sequence
changes, each of which generates more sequences than are
present in the germline genes.
5/13/2014
41
Mechanisms of diversity in antigen receptors

5/13/2014
42
Maturation and selection of

B lymphocytes
Steps in the maturation and selection of

B lymphocytes (Occur in Bone marrow)
5/13/2014
43
Maturation and selection of

T lymphocytes
The process of T lymphocyte maturation are largely related to
the specificity of different subsets of T cells for peptides
displayed by different classes of MHC molecules.
The most immature progenitors are called pro-T cells or
double-negative T cells because they do not express CD4 or
CD8.
These cells expand in number mainly under the influence of
IL-7 produced in the thymus.
Some of the progeny of double-negative cells undergo TCR
gene recombination, mediated by the V(D)J recombinase.
5/13/2014
44
5/13/2014
Steps in the maturation and selection of major

histocompatibility S.Wahyuni/lecture/immunology/S1
complex (MHC)-restricted T lymphocytes
45
SUMMARY
In the adaptive immune system, the molecules responsible for
specific recognition of antigens are antibodies and T cell
antigen receptors.
Antibodies (also called immunoglobulins) may be produced as
membrane receptors of B lymphocytes and as proteins
secreted by antigen-stimulated B cells that have differentiated
into antibody-secreting plasma cells. Secreted antibodies are
the effector molecules of humoral immunity, capable of
neutralizing microbes and microbial toxins and eliminating
them by activating various effector mechanisms.
TCRs are membrane receptors and are not secreted.
5/13/2014
46
The core structure of antibodies consists of two heavy chains

and two light chains forming a disulfide-linked complex. Each
chain consists of a variable (V) region, which is the portion
that recognizes antigen, and a constant (C) region, which
provides structural stability and, in heavy chains, performs the
effector functions of antibodies.
T cell receptors consist of an chain and a chain. Each chain
contains one V region and one C region, and both chains
participate in the recognition of antigens, which for most T
cells are peptides displayed by MHC molecules.
The V regions of Ig and TCR molecules contain hypervariable
segments, also called complementarity-determining regions,
which are the regionsS.Wahyuni/lecture/immunology/S1
of contact with antigens.
5/13/2014
47
The genes that encode antigen receptors consist of multiple segments that
are separate in the germline and are brought together during the
maturation of lymphocytes. In B cells, the Ig gene segments undergo
recombination as the cells mature in the bone marrow, and in T cells the
TCR gene segments undergo recombination during maturation in the
thymus.
Receptors of different specificities are generated in part by different
combinations of V, D, and J gene segments. The process of recombination
introduces variability in the nucleotide sequences at the sites of
recombination by adding or removing nucleotides from the junctions. The
result of this introduced variability is the development of a diverse
repertoire of lymphocytes, in which clones of cells with different antigen
specificities express receptors that differ in sequence and recognition, and
most of the differences are concentrated at the regions of gene
recombination.
5/13/2014
48
During their maturation, lymphocytes undergo alternating cycles of

proliferation and antigen receptor expression and traverse several
checkpoints at which they are selected such that only cells with complete
functional antigen receptors are preserved and expanded. In addition, T
lymphocytes are positively selected to recognize peptide antigens
displayed by self MHC molecules.
Immature lymphocytes that strongly recognize self antigens are negatively
selected and prevented from completing their maturation, thus
eliminating cells with the potential of reacting in harmful ways against self
tissues.
5/13/2014
49
Review questions
What are the functionally distinct domains (regions) of
antibody and TCR molecules? What features of the amino acid
sequences in these regions are important for their functions?
What are the differences in the types of antigens recognized
by antibodies and TCRs?
What mechanisms contribute to the diversity of antibody and
TCR molecules? Which of these mechanisms contributes the
most to the diversity?
What are some of the checkpoints during lymphocyte
maturation that ensure survival of the useful cells?
What is the phenomenon of negative selection, and what is its
importance?
5/13/2014
50

4 - Antigen Recognition - 3ed (Compatibility Mode)

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

4 - Antigen Recognition - 3ed (Compatibility Mode)

Uploaded by

Copyright:

Available Formats

ANTIGEN RECOGNITION IN THE

ADAPTIVE IMMUNE SYSTEM

The principal function of cellular receptors is to detect external

Antigen Receptors of Lymphocytes

Antigen receptor molecules consist

Antigen receptors are noncovalently attached to other

Properties of antibodies and T cell antigen receptors (TCRs).

The structure of antibodies: immunoglobulins G (IgG) and M (IgM).

Regions of antibody based on the properties

Features of the major isotypes (classes) of antibodies

Antibodies are capable of binding a wide variety of

Binding of an antigen by an antibody

Most of antibody monoclonal are made by fusing cells

B cell receptors for antigens

T cell receptors for antigens

4-5 The structure of the T cell antigen receptor (TCR)

and the chain of the TCR participate in specific

The recognition of a peptide-MHC complex

Features of antigen recognition by immunoglobulins (Igs) and

Development of Immune Repertoires

Steps in the maturation of lymphocytes

Antigen receptors are encoded by several gene

Maturing lymphocytes are selected at multiple steps

Production of diverse antigen receptors

The somatic recombination of V and J, or V, D, and J, gene

Diversity of antigen receptors is produced by:

Mechanisms of diversity in antigen receptors

Maturation and selection of

Steps in the maturation and selection of

Maturation and selection of

Steps in the maturation and selection of major

The core structure of antibodies consists of two heavy chains

During their maturation, lymphocytes undergo alternating cycles of

You might also like