B Cell Development

Rodney DeKoter
Immunology 3300B
February 9, 2015

Learning Objectives for This Lecture

1) To learn the pathways of blood cell development

2)

To learn the stages of B cell development

3) To learn how the stages of B cell development relate

to immunoglobulin gene rearrangement and clonal selection
4) To learn the stages of B cell maturation in peripheral lymphoid organs

Readings for Lecture 16: B Cell Development

Chapter 10

Growth Factor Receptors Involved in Blood Cell Development

Overview of B Cell Development

BCR

Pre-BCR
CD19

B220
IgH V-(D)J

HSC

Pro-B

Large Pre-B

bone marrow

Loss of
IL-7R

Ig V-J

Small Pre-B

Immature
B Cell,
Mature
B Cell
blood

Generation of Progenitor B (Pro-B) Cells

CD19

HSC

B220

Pro-B
earliest committed stage of B cell development
marked by expression of B220, Pax-5, CD19, RAG genes, and TdT
high levels of IL-7/IL-7R/c signaling
immunoglobulin genes are poised for recombination
DH - JH recombination initiates

Generation of Pre-B Cells (1)

Pre-BCR
CD19

B220
IgH V-(D)J

HSC

Pro-B

Large Pre-B
Functional VH - DHJH rearrangement and surface expression
of a pre-BCR marks the pre-B cell stage
VpreB and 5 proteins together assemble with heavy chain
To form a structure that resembles surface IgM
Pre-BCR is thought to signal without requirement for ligand.
Signal may control allelic exclusion.

The Pre-BCR is an important

Quality control mechanism
For B cell development

Generation of Pre-B Cells (2)

Pre-BCR
CD19

B220
IgH V-(D)J

HSC

Pro-B

Large Pre-B

Loss of
IL-7R

Small Pre-B

After a number of cell divisions, the surrogate light chain

proteins are diluted out and signaling stops. IL-7R is
shut off and cells stop dividing.
Block in mitosis induces high-level expression of
RAG-1 and RAG-2
Light-chain genes initiate recombination.

Generation of Immature B Cells

BCR

Pre-BCR
CD19

B220
IgH V-(D)J

HSC

Pro-B

Large Pre-B

Loss of
IL-7R

Ig V-J

Small Pre-B

Immature/T1
B Cell

cell-surface expression of IgM (heavy and light chains)

marks the immature B cell stage
the ratio of Ig+ to Ig+ B cells is 10:1 in mice, 6:4 in human
immature B cells enter blood and migrate to lymphoid organs
Cells entering lymphoid organs called transitional-1 (T1) cells

Ig Rearrangement Take Place At Distinct Stages of B Cell Development

Multiple rearrangements are possible at the Ig light chain loci

B Cell Development is a Wasteful Process Characterized

by Numerous Life versus Death Checkpoints

Negative Selection During B Cell Development

Negative Selection During B Cell Development

young adult mice produce 2 x 107 immature (IgM+, IgD-) B cells/day
only 5-10% of these will ever enter the pool of long-lived mature cells.
What happens to the other 90-95%?
1) Deletion of autoreactive B cells.
40-60% of all newly generated immature B cells have antigen
receptors that recognize self-antigens.
-high-affinity self-reactivity results in cell death
-also known as central tolerance
-intermediate self-reactivity results in receptor editing
Receptor editing: the re-induction of RAG recombinases
to allow one last round of light chain V(D)J recombination.
2) Exclusion of low-quality B cells.
Experiments suggest that immature B cells expressing very high
levels of surface IgM are preferred. B cells expressing low
levels of surface IgM are excluded from the spleen

Mature B Cells
BCR

Pre-BCR
CD19

B220
IgH V-(D)J

HSC

Pro-B

Large Pre-B

Loss of
IL-7R

Ig V-J

Small Pre-B

IgM and IgD are coexpressed in mature B cells, using

a RNA differential splicing mechanism.

Immature/T1
B Cell
IgM/IgD

T2/Mature B cells are competent to proliferate in response

to antigens and cytokine signals
T2 B Cell

Mature B Cells Enter The Spleen

B Cell Maturation in The Spleen

sIgM

sIgM, sIgD

sIgM, sIgD

CD21
CD93

CD23

CD93

Immature
B Cell

Transitional
B Cells

Mature
B Cells

BAFF / BAFFR signaling

CD93 = C1q-like receptor

CD21 = complement receptor 2
CD23 = low affinity IgE receptor

Summary
1) Lymphoid progenitor cells are derived from hematopoietic stem
cells and are dependent on cytokine signaling for development.
2) Immunoglobulin rearrangement occurs at distinct stages during
B cell development
3) B cell development is a wasteful process characterized by numerous
life versus death checkpoints
4) Immature B cells migrate to the spleen to complete their maturation
5) Mature B cell subsets include B-2 (follicular, marginal zone) and B-1 cells

Readings for Lecture 17: Primary Immunodeficiency

Chapter 10 review
Chapter 18 pages 593 - 599