DR / A.

S
01556364241

General Microbiology
& Immunology

Lec-11
Immunology

Level -2
Pharm -D
 B cell-mediated immune response

• naive B cells circulate throughout the body via the lymphatic system.
• These cells produce antigen-specific molecules that are necessary for detecting infectious pathogens in the human body.
• When naive B cells in the lymphatic system come into contact with an antigen, they begin the differentiation process that results in the formation of memory
B cells and effector B cells.
• Memory B cells and effector B cells produce the same antigen-specific molecules as their parent naive B cell during this development.
• The activated memory B cells express these antigen-specific molecules on their surface with the help of T cell lymphocytes, which are activated by MHC
class II receptors that recognize microbial-associated antigens.
• The effector B cells secrete these molecules in the blood to bind the antigen of interest.

✓ naive B-cells exit bone marrow, migrate to secondary lymphoid organs, then express both
surface IgM and IgD
✓ naive B-cells recirculate between blood and lymphoid organs, entering B cell follicles in
B-Cell Activation lymph nodes and spleen, responding to antigen encounter with T cell help, leading to
antibody production.
✓ B cells then interact with exogenous antigen and/or T helper cells
➢ antigen dependent phase

B-cells are activated by two mechanisms:

(Antigen dependent phase)
1-T- independent B-cell responses:
➢ Direct recognition of antigen (T-
independent antigen),
➢ T-independent antigens: are antigens
Mechanism of which can directly stimulate the B-cells
Activation
to produce antibody without the
requirement for T cell help.
➢ In general, polysaccharides are T-
independent antigens. (B-cell epitopes)
➢ Its hallmarks are to be rapid (days), while
eliciting the transient (months) production of antibodies of low affinity without inducing immune memory.
 2-T- dependent B-cell responses:
➢ Through the help of T cell (in presence of T- dependent antigen).
➢ T- dependent antigens: are those that do not directly stimulate the
production of antibody without the help of T cells.
➢ Proteins are T-dependent antigens. (T-cell epitopes)
➢ T cell dependent activation of B cells supports the generation of
two types of cells:
1) Plasma cells that secrete high affinity antibodies and
2) Memory cells that may be converted into plasma cells at a
later time Its hallmarks are to be slow (weeks), while eliciting
long-lasting (years) production of antibodies of high affinity
Mechanism of and immune memory.
Activation

Isotype (class) switching: Class switching recombination (CSR)

✓ A biological mechanism that changes a B cell's production of antibody from one class
to another; for example, from an isotype called IgM to an isotype called IgG.
✓ During this process, the constant region portion of the antibody- heavy chain is
changed, but the variable region of the heavy chain stays the same.
✓ Since the variable region does not change, class switching does not affect antigen
specificity. Instead, the antibody retains affinity for the same antigens, but can
interact with different effector molecules.
 Important definitions
Antigen (Ag) or A foreign substance that can induce specific immune response and reacts with the products of a specific immune response. Ag
Immunogen: characterized by immunogenicity and immunoreactivity.
Immunogenicity: The ability of an antigen to stimulate the immune system to induce a specific immune response
Immunoreactivity: The ability of an antigen to combine with corresponding antibody of sensitized T cell

Are small molecules which could never induce an immune response when
administered by themselves but which can when coupled to a carrier molecule
Haptens Incomplete
antigen (posses only immunoreactivity but has not immunogenicity), Ex: Penicillin
 Antigen determinants the portion of antigen molecules which can be specifically bound by antibody or lymphocyte receptor (TCR or BCR).
(epitopes):

Glycoprotein molecules which are produced by plasma cells in response to an immunogen

Antibodies or (antigen).
immunoglobulins (Igs): The portion of antibody that binds to epitope is called antigen binding site (paratope)

▪ The valency of antibody refers to the number of antigenic determinants (epitopes) that an individual antibody molecule can bind.
Valency ▪ The valency of all antibodies is at least two and in some instances more.

Antibodies Immunoglobulins

Basic structure of immunoglobulins

Although different immunoglobulins can differ structurally they all are built from the same basic unit (monomer).

• All immunoglobulins have a four chain structure as their basic unit. They are
composed of two identical light chains (23 kDa) and two identical heavy
chains (50-70 kDa).
• There are five types of Ig heavy chain (in mammal) denoted by the Greek
A. Heavy and Light letters: α, δ, ε, γ, and μ.
Chains • There are two types of Ig light chain (in mammal), which are called lambda
(A) and kappa (k).
 1. Inter-chain The heavy and light chains and the two heavy chains are held
together by inter-chain disulfide bonds interactions. The number of inter-
B. Disulfide bonds chain disulfide bonds varies among different immunoglobulin molecules.
2. Intra-chain Within each of the polypeptide chains there are also intra-chain
disulfide bonds.

Both the heavy and light chain could be divided into two regions based on variability in the amino
C. Variable (V) and
acid sequences:
Constant (C) Regions
1. Light Chain - VL (110 aa) and L (110 aa)
2. Heavy Chain - VH (110 aa) and CH (330-440 aa)

The region at which the arms of the antibody molecule forms a Y is called the hinge region because there is some flexibility in the
D. Hinge Region molecule at this point.

Carbohydrates are attached to the CH2 domain in most immunoglobulins.

E. Oligosaccharides However, in some cases carbohydrates may also be attached at other locations

Structural Regions
 Immunoglobulin fragments
Structural/function relationships
Immunoglobulin fragments produced by proteolytic digestion have proven very useful in elucidating structure/function
relationships in immunoglobulins.

▪ Fragment of Ag binding
▪ Digestion with papain breaks the immunoglobulin molecule in the hinge region before
1- Fab the H-H inter-chain disulfide bond. This results in the formation of two identical
fragments that contain the light chain and the VH and CHI domains of the heavy chain.
Two units monovalent

▪ Fragment of constant
region (crystallized)
▪ Digestion with papain
also produces a
fragment that contains
2. Fc the remainder of the
two heavy chains each containing a CH2 and CH3
domain. This fragment was called Fc because it
was easily crystallized.
▪ Effector functions The effector functions of immunoglobulins are mediated by this part of the
molecule. Different functions are mediated by the different domains in this fragment
 ▪ Treatment of immunoglobulins with pepsin results in cleavage of the heavy chain after the H-
H inter-chain disulfide bonds resulting in a fragment that contains both antigen binding sites.
▪ This fragment was called F(ab')2 because it was divalent.
▪ The Fe region of the molecule is digested into small peptides by pepsin. The F(ab')2 binds
antigen but it does not mediate the effector functions of antibodies.

3. F(ab')2

General function of immunoglobulins

▪ Immunoglobulins (Ig) bind specifically to one or a few closely related antigens.
1. Ag binding ▪ Each immunoglobulin actually binds to a specific epitope or antigenic determinant (immunologically active region on a complex antigen
that actually bind to Ig, TCR, BCR).

▪ Often the binding of an antibody to an antigen has no direct biological effect. The
significant biological effects are a consequence of secondary "effector functions" of
antibodies.
2. Effector Functions ▪ The immunoglobulins mediate a variety of these effector functions. Usually the ability to
carry out a particular effector function requires that the antibody binds to its antigen.
Not every immunoglobulin will mediate all effector functions. Such effector functions include
1- Fixation of complement: This results in lysis of cells, release of biologically active molecules
2- Binding to various cell types
▪ Phagocytic cells. NK, mast cells, and basophils have receptors that bind immunoglobulins and the binding can
activate the cells to perform some function.
▪ Some immunoglobulins also bind to receptors on placental trophoblasts. The binding results in transfer of the
immunoglobulin across the placenta and the transferred maternal antibodies provide immunity to the fetus and
newborn

Detailed function of immunoglobulins

1. Complement ▪ Activation of the complement cascade (classical pathway) by antibody can result in lysis of certain bacteria and viruses.
activation

▪ Neutralizing antibody, or NAb is an antibody which

defends a cell from an antigen or infectious body
by inhibiting or neutralizing any effect it has
biologically.

▪ They have the ability to fight antigen which attack

the immune system, since they can neutralize
2. Neutralization function without a need for white blood cells. For
example:

▪
Antibody to diphtheria toxin (antitoxin) can
prevent the binding of the toxin to host cells
thereby neutralize the biological effects of
diphtheria toxin
 ▪ Opsonization involves the binding of an opsonin, e.g..
antibody, to a receptor on the pathogen's cell
3. Opsonization membrane. After opsonin binds to the membrane,
phagocytes are attracted to the pathogen.

▪ ADCC occurs when antibody forms abridge between an infected

4. Antibody-dependent target cell (virus infected cells of the host or tumor cell) and an
cellular cytotoxicity FcR-bearing effector cell, particulary natural killer cells.
(ADCC) ▪ This results in the death of target cell either by lysis or
apoptosis.