Lecture 4 - Immunology

Antibody
Lymphoid organs
MHC
 What is an antibody?
Produced by Plasma cell (B-lymphocytes producing Ab)

Essential part of adaptive immunity

Specifically bind a unique antigenic epitope (also called an

antigenic determinant)

Possesses antigen binding sites

Members of the class of proteins called immunoglobulins

 What does an antibody look like ?
Variable region

2 identical heavy chains

2 identical light chains

Each heavy chain has a

Constant region

constant and a variable

L L region

Each light chain has a

constant and a variable
H H region
Antibody: structure and function

Fab fragment antigen

binding

Fc- Fragment constant

Antibody: Fab
Fab region
Variable region of the
antibody

Tip of the antibody

Binds the antigen

Specificity of antigen
binding determined by
VH and VL
Antibody: Fc
Fc region
Constant region

Base of the antibody

Can bind cell receptors

and complement
proteins
 Antibodies exist in two forms
Antibodies occur in 2 forms

Soluble Ag: secreted in blood and tissue

Membrane-bound Ag: found on surface of B-cell, also known

as a B-cell receptor (BCR)
 Antibody: 5 classes (isotypes)
There are 5 classes/isotypes of antibodies (IgM, IgG , IgD,
IgA and IgE).

In any given class of antibody, the constant region

contains one of five heavy-chain sequences (, ,, , or
) called isotypes

The heavy-chains determines the class of an antibody (,

IgM; , IgG; , IgD; , IgA; and , IgE).
 Immunoglobulin G (IgG)
2 Heavy and 2 Light chains Fab
(Monomer) VH
VL
~75% of serum CH1
immunoglobulin. CL

The major antibody of the CH2

secondary immune Fc
response (re-infection with
the same pathogen) CH3
Immunoglobulin M (IgM)
10 Heavy and 10 Light chains
(Pentamer)

<10% of serum immunoglobulin.

May be secreted in saliva / milk

Single J Chain (15 kDA) helps in

polymerization

J-Chain
First Ab to be produced by virgin B
cells when stimulated by an antigen;
First Ab in primary infection
Immunoglobulin A (IgA) in serum

Two forms

Monomeric form is
present in serum

~15 % of serum
immunoglobulins
Immunoglobulin A (IgA) in secretions
J-Chain

Secretory Component

dimeric
J-Chain and secretory component
Predominant antibody in secretions - saliva /milk
Not found in serum;
Confers mucosal immunity
Immunoglobulin E (IgE)

Monomeric

Trace serum protein;

bound to mast cells

Associated with allergies

(hypersensitivity)
IgE in allergy
IgE helps eliminate parasites
Immunoglobulin D (IgD)

Monomeric

Serves as a membrane
receptor on B lymphocytes
Mature B-lymphocytes are coated
with IgM and IgD
 Antibody diversity
The B cells produce millions of different types of antibodies

If 1 gene encoded 1 immunoglobulin of a given specificity, approximately

107 - 108 genes would be required to encode JUST antibodies.

However, we have only about 30,000 genes in total in the human genome

This number of genes cannot encode ALL antibody specificities (~107

108)

How is such antibody diversity possible ?

 Antibody diversity
http://garlandscience.com/garlandscience_res
ources/resource_detail.jsf?landing=student&r
esource_id=9780815342434_CH05_QTM01
 VDJ recombination and Ab diversity
Heavy chain Light chain
V 1000 300
D 15 -
J 4 4
VxDxJ VxJ
1000 x 15 x 4 300 x 4
= 6 X 104 1.2 x 103
X
= 7.2 x 107

One DNA sequence millions of Abs

Lymphatic system
What is the lymphatic system ?
Part of the circulatory system

Removal of interstitial fluid

Lymphatic system: What is it made of ?

Lymphatic vessels

Lymph nodes

Lymphatic tissue

Organs
 Primary lymphoid organs
Bone marrow:
Makes all the blood cells including lymphocytes
Removes auto-reactive (B cells that recognize self) B-
lymphocytes

Thymus:
Removal of auto-reactive (T cells that recognize self) T-
lymphocytes
 Removal of auto-reactive B cells in the
bone marrow

Self-antigens

Immature B-cells

Mature B-cells

Apoptosis (happens to 90% of the B cells)

 Removal of auto-reactive T cells in the
Thymus

Self-antigens

Immature T-cells

Mature T-cells

Apoptosis
 Immunological tolerance
Non-responsiveness of the immune system to self
antigens or substances capable of eliciting an
immune response.

This is used by the immune system to discriminate

self from non-self
 What is the function of the lymphatic
system?
Lymphatic vessels
Removal of excess interstitial fluid
Transport lymphocytes

Primary Lymphoid organs (BM, Thymus)

Production of lymphocytes
Removal of auto-reactive lymphocytes

Secondary Lymphoid organs (L.nodes, tonsils, spleen..)

Collection of lymphocytes (Antigen presenting cells /
macrophages..) to fight infection
MHC (Major histocompatibility
complex)
 What are MHC ?

They are glycoproteins that serve as antigen

presenting structures on cell surface

Essential part of adaptive immune response

 MHC class I

Present on almost all cells

Present antigens to CD8+ T

cells (cytotoxic T cells)

Infected host cell

 MHC class II

Present on cells of the

immune system - eg.
macrophages, B cells

Present antigens to CD4+ T

cells (T helper cells)
 MHC class I vs MHC class II

MHC class I MHC class II

Expression Almost all cells Only some cells of the
immune system:
macrophages, B cells etc
Antigen presentation To CD8+ T cells To CD4+ T cells
Nomenclature A, B, C, D, E, F DP, DQ, DR

MHC = HLA or human leukocyte antigen

 MHC Diversity

MHCs are highly polymorphic

MHCs are isoantigens

MHC I - >1300 variants

MHC II - > 700 variants

MHC-peptide binding is promiscuous binding

There are hundreds of thousands of antigens but only a

limited number of MHCs

To be able to present all/most antigens MHCs bind

promiscuously to peptides

1 peptide binds to many MHCs

1 MHC binds to several peptides
What will happen if we all have the same
MHC make-up ?

Threat to individuals survival Possible extinction of population

 MHC diversity and disease outcomes

Impact on individual depends on MHC type Population survives

T cells and B cells
 Lymphocyte subsets

CLP
T CELLS B CELLS
Common
lymphoid
T precursor B

Th CTL PC
T HELPER CELLS CYTOTOXIC T Produce antibodies
(TH Cell or CD4+ T cells) LYMPHOCYTES PLASMA CELLS
(Tc Cell or CD8+ T cells)
Lymphocyte antigen receptors

The B cell antigen receptor is a membrane-bound antibody

B SURFACE IMMUNOGLOBULIN (BCR = B cell receptor)

The T cell antigen receptor IS NOT an antibody but a distinct

T molecule
T CELL receptor (TCR)

Each antigen receptor binds to a different antigen

Each cell has only one antigen specificity
 B cell and T cell receptors

TCRs also undergo VDJ recombination similar to Abs millions of TCRs

T helper cell (TH or CD4+ T cell) activation
A closer look at T helper cell activation

APC

TCR to MHC on APC Are you me ? - YES

Signal 1
TCR to Antigen Are you me ? - NO

TH Cell to APC Do you have B7 co-stimulatory signal ? - YES Signal 2

Signal 1 + Signal 2 activate T cell

 What happens when a T helper cell
(TH Cell) is activated ?

1
2

(1) Proliferation of TH cell (CD4+ T cell)

(2) Proliferation of CTL (CD8+ T cell)
(3) Activation of B cell to produce antibody
What happens when CD8+ cells (Cytotoxic T
cells or CTLs) recognize infected cells ?

Cytokines
Activated TH cell

Activated CTL
Killing of host cells
release perforins
How does B-cell handle a pathogen ?
TH cell help to B-cell

I can produce antibodies

 What happens when a B cell is
activated ?

2 NO Ab
Long lived
1

Produces Ab
(1) B cells becomes a plasma cell to produce antibodies short lived

(2) B cells becomes a memory B cell

 Why do we need memory B cells?
Hey . I think I
Memory B cells know you.
Killing of
pathogen in
quick time

NO Lag time

Plasma cells

When the body encounters the same pathogen again it responds very quickly
Are there memory T cells Yes both memory TH cells and memory CTLs( but this will not be discussed)
What is the role of antibodies in immunity ?

1) Prevents virus entry into host cell

2) Promotes phagocytosis of pathogens

3) Promotes killing of infected host cells

 Antibody: (1) Virus neutralization
Virus attaches to receptor in host cell Ab-bound virus unable to attach to receptor

Virus enters the host cell Virus cannot enter the host cell
Antibody: (2) opsonisation and phagocytosis

Killing of
microbe

Lysis of
microbe by
MAC
Antibody: (3) antibody-dependent cell-
mediated cytotoxicity (ADCC)

Killing of
microbe
Summary: adaptive immune response
 T regs
T regulatory cells (Old term suppressor T cells) suppress
immune response

CD4+ (like T helper cells) but have other surface markers.

Prevent autoimmunity

Mice without Tregs die prematurely of autoimmune

diseases
 T regs

T regs suppress immune response and prevent auto immune diseases

 Antibody class switch
T Helper
cell

B cell
activation

Plasma
cells

Programming switch in B cells differentiate

IgM antibodies into plasma cells that make IgG
 Ab response and class switch

Lag phase
Log phase Ab Titer 1o Ag 2o Ag

Plateau phase
Decline phase

Days After Immunization

 Ab response and class switch

Total IgM Ab IgG Ab

Ab class switch Ab

1o - IgM
2o - IgG
Ab Titer
1o Ag 2o Ag

Days After Immunization

IgG antibody circulate in body for years - protect against specific antigens
 Hypersensitivity

INAPPROPRIATE IMMUNE RESPONSE TO AN ANTIGEN

Class I hypersensitivity

Often treated with anti-histamines

 Other classes of hypersensitivity

Class II-IV

Involve IgM, IgG, Ag-Ab complexes, c` activation

Not discussed
 Autoimmunity
Immune response against ones own-self
Possible causes of autoimmunity ?
1) Release of Sequestered Antigen

2) Cross-reactive Autoantigens (Molecular Mimicry)

3) Hormones

4) Drugs
 1. Sequestered Antigen
Some antigens are hidden from the immune system

Damage to these organs causing exposure of these

sequestered antigens an immune reaction to these
antigens may occur.

Examples
Exposure of Eye lens protein after trauma
 2. Cross-reactive or molecular Mimicry

Viruses and bacteria may have antigenic determinants that

are identical or similar to those of normal host cells

e.g. Rheumatic fever - Streptococci and proteins of the heart

muscle
Molecular mimicry and autoimmunity
Ab against streptococci can cause
rheumatic heart disease
 3. Hormones

About 75 percent of autoimmune diseases occur in

women,

most frequently during the childbearing years.

Estrogen (a hormone) is believed to play a role in

several autoimmune diseases by modulating
immune responses
Autoimmune disease affect women more
often then men
 4. Drugs
Drugs = not antigens = no immune response

Self-antigens = no immune response

Drug + self-antigen = neoantigen = immune response

Examples of autoimmune
diseases
Systemic Lupus Erythematosus (SLE)

AutoAb to nuclei, DNA

Ag-Ab complexes are

deposited in kidneys and
vascular tissue.

Joint pain, Hemolytic

anemia, Kidney
dysfunction, skin rashes
 Multiple Sclerosis
MS patients can have autoantibodies which are responsible
for the demyelination

Mild- numbness of the limbs

Severe- Paralysis or loss of vision

etc.