Lecture 15

B Cell Activation and

Antibody Production
 Overview of B Cell
Development,
Activation,
Antibody Production
 B Cell Antigens

Antigens

Thymus Dependent Antigens Thymus Independent Antigens

Dependent Upon Helper T Cells Does Not Need Helper T Cells

to Induce Antibody Production to Induce Antibody Production

Proteins Polysaccharides, Lipids

 B Cell Responses to Thymus-
Dependent Antigens (T Cell-
Dependent Antibody Responses)
Primary and Secondary Antibody Responses
Phases of the Humoral Immune Response
A T-Dependent Antigen Must Contain Both
B and T Cell Epitopes

Antigen

T Cell Epitope B Cell Epitope

LINKED RECOGNITION
Follicles

(B Cells)

(T Cells)
Activation of B Cells by Antigen and Complement

1. Biochemical Signals
2. Endocytosis of Antigen
Antigen Recognition Phase of T-Dependent
Antibody Response
Interactions of B Cells with Helper T Cells
TEM Picture

B Cell B Cell

T Cell
Initial Contact T-B Conjugate

Note the broad area of membrane contact between

B and T Cells.
Helper T Cell-Dependent Activation
Of B Lymphocytes
 B-Cell Activation by
Thymus-Dependent Antigens

C’R Cytokines

Linked Recognition
 Activated B Cells
(Following Interaction with TH Cells

Extra-follicular Site Follicle

Germinal Center
Antibody Secreting
Cells

Antibodies
Late Events in T Cell-Dependent
Antibody Responses-Germinal
Center Reaction

• Affinity Maturation
– Somatic Hypermutation
• Generation of Memory B Cells
 Somatic Hypermutation and Affinity
Maturation of Antibodies

Affinity maturation is the process that

leads to increased affinity of antibodies
for a particular antigen as a result of
somatic mutation in the Ig genes followed
by selective survival of B cells producing
the antibodies with the highest affinity
Affinity Maturation in Antibody Responses
Selection of High Affinity
B Cells in Germinal Center
Phases of the Humoral Immune Response to
T-Dependent Antigen
Anatomy of Humoral Immune Responses
Antibody Isotype
Switching
Isotype Switching Under the Influence of Helper
T Cell-Derived Cytokines
Mechanism of Ig Isotype Switching
CD4 T Cell-Dependent Effects in
Antibody Responses

• Memory B Cell Development

• Isotype Switching
• Affinity Maturation
 Thymus-
Independent
Antigens
 B-Cell Activation by
Thymus-Independent and Dependent Antigens

Most TI antigens are polyvalent and induce maximal

Crosslinking of membrane Ig on B cells, without a
Need for T cell help.
 Features of Antibody Responses to
T-Dependent and T-Independent Antigens
Antibody Response to T-Dependent
Antigens

• Role of Helper T Cells

– Cytokines
– CD40/CD40L interactions
• Isotype Switching
– Switch Recombination
– Cytokines and Isotypes
• Affinity Maturation
– Somatic hypermutation
– Selection for B cells which produce High Affinity
Antibodies

• Memory B Cells
Antibody Effector
Functions
Effector Functions of Antibodies
Neutralization of Microbes by Antibodies
Neutralization of Toxins by Antibodies
Opsonization of Microbes by Antibodies
Antibody-Dependent Cellular Cytotoxicity (ADCC)
Functions of Complement
Complement-Mediated Lysis of E. coli
Alive Killed
Cellular Interactions in Immune Responses
The Immune
Response:
A Summary
WHY can immune system recognize so
many different epitopes??

Antibody heavy and light chains are

composed of gene segments
Variable regions are unique
A limited variety of constant region sequences
are used

They must be rearranged into functional genes

before they can be transcribed
p. 106
Organization of Ig genes

Germline DNA- gene segments surrounded by

noncoding regions
These are rearranged to form functional genes

Light chains- V, J and C segments

Heavy chain- V, D, J, C

V regions rearrange first

A single V can rearrange to more than one C

Multigene families

 or 

In humans: 40 V, 5 J, 1 C

Similar number of  genes in humans;

this is rare in mice

Heavy-chain gene families are similar but more

complex (D segment)

CH regions formed from exons

p. 111

One of many possible combinations

Heavy chain DNA

D-J and V-DJ rearrangements must occur

separately

On a mature B cell, both mIgM and mIgD

are expressed on the cell surface
How does rearrangement occur?

Each V, D and J is flanked by RSS

(Recombination signal sequences)

Mechanism is controlled by RAG-1 and RAG-2

proteins and an enzyme TdT

If any of these proteins is defective no mature

B cells can form; nor T cells
p. 112
“Junctional flexibility” contributes to diversity

p. 115

But not all rearrangements are “productive”

B cells are diploid and contain chromosomes
from both parents

However, heavy chain genes are rearranged

from only one chromosome, as are light chain
genes.
Therefore, any one B cell will contain
one VH and one VL (antigen specificity)

How? Allelic exclusion (Yancopoulos and Alt, 1986)

Model for allelic exclusion

p. 116
Generation of antibody diversity
(why are there so many possible antigen
combining sites?)
Multiple germline gene segments

In human germline:

51 VH, 27 D, 6 JH

40 V, 5 J 

30 V , 4 J
Combinatorial V-J and V-D-J joining

57 V X 27 D X 6 J= 8262 possible combinations

for VDJ joining

40 V X 5J = 200 possible V 120 possible V

8262 X (200+120) = 2.64 X 106 possible

combinations

Without taking into account other sources of

diversity
Junctional flexibility in V-J or V-D-J junction

Additional nucleotides added at junctions

(P or N addition), if a single-stranded
region is created during the joining
process

Somatic hypermutation
mutations occur AFTER rearrangement
tends to occur in CDR regions
affects antigen affinity (tends to increase):
“affinity maturation”
occurs in B but not T cells
Class switching

After antigen stimulation heavy-chain DNA can

rearrange so VDJ joins to any isotype

Cytokines help determine the isotype

IgG2a or IgG3 (mice): IFN-

IgM: IL-2, IL-4, IL-5
IgE: IL-4
p. 122
Membrane-bound or secreted?

Alternative splicing, p. 124

Mature B cells express both mIgM and mIgD

No switch site between C and C

The VDJCC contains 4 polyadenylation sites

mIgM or mIgD can be generated depending

on which polyadenylation site is used
Regulatory elements of transcription

Promoters

Enhancers

Gene silencers

Gene rearrangement brings enhancers close

to the promoter they influence
Why aren’t Igs produced in B cells?

In T cells a protein may bind to the -enhancer

and prevent V-J joining

Arrangement of immunoglobulin genes (and

formation from exons) and greatly
facilitated formation of genetically
engineered antibodies