Development of B Lymphocytes

Parham Chapter 6
Jim Drake
MS-449
2-9337

Learning Objectives
To be able to explain:
The role of the bone marrow stroma in B cell development
How V(D)J recombination is coordinated with B cell development
The check points in B cell development
The role of antigen in B cell development and activation
Allelic exclusion
The role of the pre-BCR in B cell development
How self-reactive B cells are removed from the developing B cell
repertoire
Receptor editing
Conversion of immature to mature B cells in peripheral lymphoid tissues
The circulation of mature nave B cells
The difference between B1 and conventional B2 B cells
The correlation between B cell tumor markers and stages in B cell
development

B Cells Birth to Death (or a long life)

Bone marrow phases 1 2

(and some of 6)
Peripheral phases 2 - 6

The Early Stages of B Cell Development

Early B cell development in bone marrow

Dependent on bone marrow stroma
Cell-cell interactions c-Kit (receptor) / stem cell factor
(SCF, membrane-bound ligand
Soluble factors IL-7 receptor / IL-7

Decreased dependence on stroma as cells differentiate

V(D)J Recombination A Review

What is V(D)J recombination?
What proteins mediate V(D)J
recombination?
What is a RSS?
What is the 12/23 rule?
Does V(D)J recombination
occur at the RNA or DNA
level?

V(D)J Recombination During

B Cell Development

Immunoglobulin heavy chain genes re-arrange before

light chain genes
Functional re-arrangement of IgH and IgL genes are
check points in B cell development
Antigen specificity unknown until both IgH and IgL genes
are re-arranged
This is antigen independent

VDJ Recombination Pro-B Cells

Heavy chain gene re-arrangement

D-J joining on both chromosomes
V to DJ joining
First try successful codes for protein
First gene expressed
Suppression of second re-arrangement

First try unsuccessful

First gene cant be expressed
Second gene re-arranged

Second try successful survival

Second gene expressed
First gene not expressed

Second try unsuccessful cell dies

Blue text allelic exclusion

(also see next slide)

The Consequence of Allelic Exclusion

Allelic exclusion each B cell
expresses only one type of
BCR (one antigen specificity)
Two mechanisms
Successful re-arrangement of
first allele blocks rearrangement of second allele
Successful re-arrangement of
second allele
Second allele expressed
First allele non-functional not
expressed

This does
not happen

Pre-B Cells The Pre-B Cell Receptor

Pre-B cell
Re-arranged IgH gene
Germ line IgL genes

Pre-BCR
Composition
Immunoglobulin light chain
Surrogate light chains VpreB, 5
CD79A-CD79B (Ig-Ig) signaling

Functions
Suppress immunoglobulin heavy chain
gene re-arrangement
Stimulate cell division
Stimulate immunoglobulin light chain
gene re-arrangement

Generation of
Immature B Cells

VJ re-arrangement of immunoglobulin
light chain genes

Ordered re-arrangement
First kappa
Then lambda
Exhibits allelic exclusion

Establishes antigen specificity of B cell /

BCR

Immature B Cells Testing for Autoreactivity

Immature B cell

IgMhi / IgDlo

BCR signaling negative event

Non-self reactive B cells

Release into periphery

Self reactive B cells

Multivalent self antigen

Receptor editing (next slide)

Apoptosis

Monovalent antigen

Anergy specific nonresponsiveness death in a

few days

Immature B Cells
Light Chain Receptor Editing
Self-reactive immature B cell
Another round of immunoglobulin
light chain gene re-arrangement

New BCR
Old IgH + new IgL
Non self-reactive B cell survives
Self-reactive
Another round of IgL rearrangement
Cell death

Early B Cell Development - Overview

Life Outside the Bone Marrow Step I

Transition from immature (IgMhi / IgDlo) B cell to mature (IgMlo / IgDhi)

in peripheral lymphoid tissue (e.g. lymph node)
Lymph node stromal cells secrete chemokines to attract B cells
Extravasation via high endothelial venules

Follicular dendritic cells (FDC) different from dendritic cells

BAFF from FDC supports B cell survival / maturation
Lymphotoxin from B cells maintain FDC network

Circulation of Nave B Cells

Nave B cell
Matured in primary follicle (previous slide) never seen antigen
Half-life of ~ 3 months (without antigen)
Circulate through secondary lymph node organs (e.g. lymph node)
Blood HEV lymph node tissue efferent lymph blood
Afferent lymph lymph node tissue efferent lymph blood

B Cell Activation
Antigen binds the BCR!
B cell T cell interactions
T cell help
Germinal centers

Development of
Antibody producing plasma cells
Memory B cells

More on this next time

Chapter 9

B1 Versus B2 B Cells

Conventional B cells B2 B cells

B1 B cells

Express CD5 marker protein

~5% of all human B cells
High degree of reactivity to carbohydrate antigens
Source of most cases of chronic lymphocytic leukemia (CLL)

B Cell Tumors
Different B cell tumors
correspond to transformed
versions of different stages
of B cell development
Knowing the hallmarks of
the stages of B cell
development will help you
classify clinical cases of B
cell tumors
Each tumor type has a
different prognosis / optimal
treatment

Learning Objectives
To be able to explain:
The role of the bone marrow stroma in B cell development
How V(D)J recombination is coordinated with B cell development
The check points in B cell development
The role of antigen in B cell development and activation
Allelic exclusion
The role of the pre-BCR in B cell development
How self-reactive B cells are removed from the developing B cell
repertoire
Receptor editing
Conversion of immature to mature B cells in peripheral lymphoid tissues
The circulation of mature nave B cells
The difference between B1 and conventional B2 B cells
The correlation between B cell tumor markers and stages in B cell
development