You are on page 1of 50

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

S.Wahyuni/lecture/immunology/S1

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
S.Wahyuni/lecture/immunology/S1
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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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
S.Wahyuni/lecture/immunology/S1

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


5/13/2014

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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


5/13/2014

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

13

Features of the major isotypes (classes) of antibodies


5/13/2014

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

15

Binding of an antigen by an antibody


5/13/2014

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

19

5/13/2014

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

23

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


5/13/2014

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

25

The recognition of a peptide-MHC complex


by a T cell antigen receptor
5/13/2014

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

28

Features of antigen recognition by immunoglobulins (Igs) and


T cell antigen
receptors (TCRs).
5/13/2014
S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

32

Steps in the maturation of lymphocytes


5/13/2014

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

37

5/13/2014

S.Wahyuni/lecture/immunology/S1

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
S.Wahyuni/lecture/immunology/S1
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

S.Wahyuni/lecture/immunology/S1

41

Mechanisms of diversity in antigen receptors


5/13/2014

S.Wahyuni/lecture/immunology/S1

42

Maturation and selection of


B lymphocytes

Steps in the maturation and selection of


B lymphocytes (Occur in Bone marrow)
5/13/2014

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

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

S.Wahyuni/lecture/immunology/S1

50