TODAY

ANTIBODY STRUCTURE AND FUNCTION

Asilmi 06 - B CELLS AND ANTIBODIES

Antigens are detected by specialized T cell receptors (TCR)
or B cell receptors (BCR).

APC

T cell B cell

T cells require antigen The BCR directly

presentation by dedicated recongnizes its
antigen presenting cells cognate antigen
(APC)

Asilmi 06 - B CELLS AND ANTIBODIES

LYMPHOCYTE ACTIVATION IS TRIGGERED BY ANTIGEN RECOGNITION

Antigen recognition is mediated MICROBE

by structurally related cell
B CELL
surface molecules: membrane-
bound antibodies on B
lymphocytes and T cell receptors
on T lymphocytes.

Antigen receptors distinguish many, and often chemically related,

structures.

Antigen receptors are clonally distributed: Each clone has a unique

receptor distinct from all other receptors

Clone: A parental lymphocyte and all its progeny.

Asilmi 06 - B CELLS AND ANTIBODIES

 BCR/TCR ACTIVATION

1. Antigen receptors are non-

Extracellular
covalently associated with
cellular signaling proteins in the

BCR COMPLEX
BCR or TCR complex.
BCR

2. When adjacent antigen

Intracellular
receptors bind two or more
antigens the receptors are pulled
together into an aggregate.

3. Cross-linking brings signaling proteins together and initiates signal

transduction.

4. As the signaling proteins are the same in each clone, the transduced
signal is also the same.
Asilmi 06 - B CELLS AND ANTIBODIES
 B CELLS

1. B CELLS ARISE FROM BONE MARROW STEM CELLS.

2. MATURE B CELLS EXIT BONE MARROW AND ENTER THE CIRCULATORY SYSTEM.

3. B CELLS RECIRCULATE THROUGH LYMPHOID ORGANS IN SEARCH OF ANTIGEN.

Asilmi 06 - B CELLS AND ANTIBODIES

 B CELLS

1. BCR detect three-dimensional

antigens.

2. B cell activation does not

require antigen presentation.

3. B cells are active only against

extracellular antigens.

Asilmi 06 - B CELLS AND ANTIBODIES

 B CELLS AND ANTIBODIES

1. Each B cell makes One specificity of antibody (Immunoglobulin (Ig)).

The antigen receptor is membrane bound on a nave B cell.

2. Ag plus second signal stimulate B cell.

3. Active B cell secretes antibody into the blood and lymph or patrolling
mucosal surfaces.
Asilmi 06 - B CELLS AND ANTIBODIES
 B CELLS AND ANTIBODIES

Secreted
antibodies

BCR

Naive
Active

In the transition from nave to active B cells, the B cells secrete a

modified version of the B cell receptor. This molecule is known as an
antibody.

The BCR and corresponding antibody share IDENTICAL antigen

specificities.

Asilmi 06 - B CELLS AND ANTIBODIES

 What are Antibodies?

Antibodies (A.K.A. Immunoglobulin) are a type of

glycoprotein produced by B lymphocytes. Antibodies bind
antigen with a high degree of specificity and affinity.
Antibodies recognize a variety of three-dimensional shapes
(amino acids, lipids, carbohydrates, etc).

Asilmi 06 - B CELLS AND ANTIBODIES

 ANTIBODY STRUCTURE
An antibody consists of four
polypeptides: two identical light
chains and two identical heavy
chains form a Y-shaped molecule.

Each light chain is connected to a

heavy chain by a disulfide bond and
the two heavy chains are connected LIGHT CHAIN

by two disulfide bonds.

Antigen binding and effector

domains are separated by a hinge
region. The hinge region allows the HEAVY CHAIN
two antigen binding domains to
move, enabling them to bind
antigens that are separated by
varying distances.
Asilmi 06 - B CELLS AND ANTIBODIES
 ANTIBODY STRUCTURE
ANTIGEN RECEPTORS HAVE
DISTINCT FUNCTIONAL DOMAINS

1. The variable (V) region varies

between clones and is involved in
antigen recognition.

2. The constant (C) region is

conserved among clones and is
required for structural integrity and
effector functions.

Asilmi 06 - B CELLS AND ANTIBODIES

 ANTIBODY STRUCTURE

The unique structural architecture of antibodies allows multiple, highly

diverse antigens to induce identical effector functions.

Antigen

Variable region

Constant region

IMMUNITY IMMUNITY IMMUNITY

Different variable regions bind different antigens. Identical constant

regions induce identical responses.
Asilmi 06 - B CELLS AND ANTIBODIES
 ANTIBODY STRUCTURE

Each Light chain contains one V

domain and one C domain.
Each Heavy chain contains one V
domain and at least three C
domains.

Each domain folds into a

characteristic 3-D shape:
The Immunoglobulin (Ig)
domain.

VL

Antibodies bind antigens through the variable

regions of the light and heavy chains.
VH
Asilmi 06 - B CELLS AND ANTIBODIES
 BINDING OF ANTIGEN BY ANTIBODY - TERMS AND CONCEPTS
Epitope
Antibodies bind antigens by reversible non-
Epitope
covalent interactions.

Epitope: The parts of an antigen recognized

by an antibody are called epitopes. Epitopes
can be recognized on the basis of sequence
or shape.

Affinity: The strength with which one

antigen-binding surface of an antibody binds
an antigen.

Avidity: Each Isotype has between 2 and 10 antigen binding sites. Therefore, each antibody
can bind 2 to 10 epitopes of an antigen, as long as identical epitopes are sufficiently close
together, e.g. microbial cell surface proteins. In this case the binding is much greater than
the affinity of a single antibody-antigen bond and is called the avidity.

Asilmi 06 - B CELLS AND ANTIBODIES

Specific Antibody (Ig) Molecules Are Tailored to Exactly Fit A
3 D Surface Site On An Antigen.

Epitope

Influenza coat protein Ig

Asilmi 06 - B CELLS AND ANTIBODIES

 FEATURES OF ANITBODY-MEDIATED
ANTIGEN RECOGNITION

1. Antibodies recognize a large array of 3-D structures.

2. Each clone is specific for a single antigen.

3. Antigen recognition is mediated by specific domains of the

antibody.
4. Signaling triggers B lymphocyte activation.

Asilmi 06 - B CELLS AND ANTIBODIES

 B CELL ACTIVATION
B cell activation requires two signals
1. Antigen
1. Antigen
A. T Cell-independent. 2. PAMP
2. PAMP
PRR PRR
1st signal: BCR detects Ag. BCR BCR
2nd signal: PRR detects PAMP.

B cell Active B cell

Cytokines
B. T Cell-dependent. 1. Ag
CD4+ T cell
1st signal: BCR detects Ag.
B cell internalizes Ag and
presents to CD4+ T cell (B cell as BCR MHC II/Ag
APC).
2nd signal: T cell produces
cytokines that activate B cell. B cell
Asilmi 06 - B CELLS AND ANTIBODIES
 IMMUNOGLOBULIN ISOTYPES
There are five types of heavy chain that differ in
their C region. Each class is an Isotype. Each
Isotype differs in their physical/biological
properties and effector functions.

IgA. Two IgA The J chain facilitates transport of IgA

molecules joined across mucosal epithelia. The J chain
by J chain. also facilitates transfer of IgA to
newborns to confer neonatal passive
immunity.

Asilmi 06 - B CELLS AND ANTIBODIES

 IMMUNOGLOBULIN ISOTYPES

IgD. Function unknown. Restricted to

membrane and not expressed on active B
lymphocytes. IgD knock-out mice do not have
any apparent defects.

Asilmi 06 - B CELLS AND ANTIBODIES

 IMMUNOGLOBULIN ISOTYPES

Antigen

IgE
Fc Receptor

Secreted as a
monomer. MAST
CELL

IgE binds the Fc receptors of mast cells.

When IgE is cross-linked by

antigen, it triggers mast cell
degranulation - allergic
response.
Asilmi 06 - B CELLS AND ANTIBODIES
IMMUNOGLOBULIN ISOTYPES
Secreted as
monomer

IgG can bind

and neutralize
toxins.

Asilmi 06 - B CELLS AND ANTIBODIES

 IMMUNOGLOBULIN ISOTYPES
Opsonization: IgG coats pathogens and
prevents them from entering host cells.

Infectious pathogen
entering host cell
to propagate

Pathogen cannot
enter cell
when coated
(opsonized) with
antibody

Asilmi 06 - B CELLS AND ANTIBODIES

 IMMUNOGLOBULIN ISOTYPES
Ag

IgG Phagolysosme
FcR

Macrophage

MHC II/peptide

1. IgG-Ag binds the Fc receptor of

macrophages.
2. Ag is internalized through phagocytosis.
3. Microbe is degraded in the phagolysosome.
4. Microbial peptides are presented on Class II
MHC molecules - CD4 T cell activation! Asilmi 06 - B CELLS AND ANTIBODIES
 IMMUNOGLOBULIN ISOTYPES
Fc receptors also mediate Antibody-Dependent
Cellular Cytotoxicity (ADCC)

Natural Killer Cells

Ig Target
Ag

1. Ig binds Ag on surface of target cell.

2. Fc Receptors on Natural Killer cells bind Fc

of Ig.

3. Cross-linking of Fc receptors signals to the

NK cell to kill the target.

4. Target cell dies by apoptosis Asilmi 06 - B CELLS AND ANTIBODIES

 IMMUNOGLOBULIN ISOTYPES
IgM exists as pentamers

10 different Ag binding
sites - very high avidity!

As with IgA, it has a J

chain for secretion.

IgM is the first antibody expressed in mature B

cells.

Important for activation of the complement

pathway. Appears early after infection and
usually not seen upon re-infection.

Asilmi 06 - B CELLS AND ANTIBODIES