The Development of B Lymphocytes

B Cells
The primary function of B cells is to produce antibodies. B-cells are derived from bone marrow precursor cells that arise continuously throughout life. B-cell development can be separated into two phases
antigen-independent phase which occurs in primary lymphoid organs (fetal liver and adult bone marrow) antigen-dependent phase which occurs in secondary lymphoid organs (spleen, lymph nodes, gut Peyers patches.

CORE MATERIAL: Four Phases of B Cell Development

Generation of B cell receptors in the bone marrow. Tolerance induction by self antigens in BM and periphery. B cell activation by foreign antigens (secondary lymphoid organs). Differentiation to antibody secreting (plasma cells) and memory cells in secondary lymphoid organs.

CONCEPT: B Cell Differentiation Proceeds Through Several Definable Developmental Stages.

Bone Marrow Bone Marrow
IgM

Periphery
Y Y Y Y Y Y Y Y

Y
Lymphoid Stem Cell Immature B Cell

IgM

IgD

Antigen
IgM

Plasma Cell ProB Cell PreB Cell Mature B Cell Activated B Cell Memory B Cell Antigen Independent

Antigen Dependent Secondary Lymphoid Organs

Primary Lymphoid Organ

CORE MATERIAL: The antigen specificity of each individual lymphocyte is determined through the assembly of V, (D), and J gene segments to generate rearranged V genes encoding the antigen-receptor varialbe (V) region. CORE MATERIAL : The rearrangement of antigenreceptor gene segments controls lymphocyte development. CORE MATERIAL: Errors in immunoglobulin-gene rearrangement can give rise to chromosomal translocations that predispose B cells carrying them to malignant transformation.

CONCEPT: B cells develop in the bone marrow and then migrate to the secondary lymphoid tissues in the periphery.

CONCEPT: The development of B cells proceeds through stages defined by the rearrangement and expression of the immunoglobulin genes.

Pro-B-Cells

Progenitor B-Cells

Pro-Bells are the stage where D to J rearrangements are carried out. Generally the cell will rearrange the D and J segments on both alleles. Once DJ rearrangements are complete the cells progress through several intermediate stages in which an ordered rearrangement of the heavy chain V region is carried out. Pro B cells express lymphoid specific enzymes, RAG1/RAG2, and TdT, as well as characteristic surface proteins.

Pre-B-Cells

Pre-B Cells

Pro-B to Pre-B-Cell transition entails V to DJ joining of the H chain on one allele. The productively rearranged immunoglobulin gene is expressed immediately as a protein by the developing B cell both in the cytoplasm and as a component of the pre-B cell receptor. Successful rearrangement of the heavy-chain immunoglobulin gene segments leads to the formation of a pre-B cell receptor that halts further VH to DJH rearrangement and triggers the cells to begin to divide. Once the developing B cells complete a productive rearrangement of the H chain locus, they begin an ordered rearrangement of the light chain loci. TdT is no longer expressed by cells at the pre-B cell stage. Pre-B cells do express RAG1/RAG2 in the nucleus.

CONCEPT: Success of the Developmental Journey, at all Steps, Depends on Signaling Through Receptors on the Surface of the Developing B Cells

Sequential Expression of Membrane Ig and Surrogate Light Chain at Different Stages of B-Cell Development

Pre-B-Cell Receptor
Pre-B-cell receptor (pre-BCR) is composed of a -chain and a surrogate light chain and the signaling molecules Ig and Ig . Surrogate L-chain complex of two proteins a V like sequence called Vpre-B and a C-like sequence called 5, which associate noncovalently to form a light-chain like structure. Gene-targeted 5 knockout mice are blocked at the pre-B cell stage, suggesting that a signal generated through the receptor is necessary for pre-B cells to proceed to the immature B-cell stage.

Immature B-Cells

Immature B Cells

Once a light-chain gene has been rearranged successfully, light chains are synthesized and combine with the heavy chain to form IgM (of a single specificity). IgM appears of the cell surface along with Ig and Ig to form the functional B cell receptor (BCR). Expression of the BCR signals the cells to down regulate RAG1/RAG/2 and to cease further rearrangements. This is the stage of development where negative selection of cells that recognize self antigens occurs. The immature B cells egress from the bone marrow and enter the circulation and ultimately arrive in the spleen. Any encounter with antigen on this journey results in initiation of the death program in the cells.

Membrane-bound immunoglobulins associate with two other proteins, Ig and Ig , to form the B-cell Receptor (BCR)

Antigen Recognition

Signal Transduction

CONCEPT: B cells undergo a strictly programmed series of gene rearrangements in the bone marrow.
The recombination process is imprecise due to the random addition and deletion of nucleotides at the joins between gene segments. Thus, by a matter of chance stop codons can be introduced which prevent synthesis of a complete chain (termed non-productive rearrangements). B-development has evolved to preserve and multiply those B cells that have made productive joins and eliminate that have not. Assembly of genes for a complete BCR requires three separate recombination events, which occur at different stages of development. These are, in order that they occur DH to JH, VH to DJH to produce the functional heavy chain gene, and the joining of VL to JL to produce the functional light chain gene. The kappa chain locus is generally rearranged before the lambda chain locus.

Summary of B-cell Development

Mechanisms of Self-Tolerance Unresponsiveness to self antigens, or selftolerance, is maintained by mechanisms that actively prevent the maturation or stimulation of potentially self-reactive lymphocytes. Several fundamental concepts are relevant to our understanding of selftolerance.

Fundamental Concepts
Tolerance to self antigens is an actively acquired process (rather than an inherited property), in which self-reactive lymphocytes either are prevented from becoming responsive to self antigens or are inactivated after encountering these antigens. Self-tolerance may be induced at various stages of lymphocyte development and activation.

Fundamental Concepts
Central Tolerance is the induction of tolerance in generative (primary) lymphoid organs as a consequence of immature selfreactive lymphocytes recognizing self antigens. The bone marrow compartment has two mechanisms for dealing with autoreactive B cells.
Clonal Deletion by apoptosis Receptor editing

Fundamental Concepts
Peripheral Tolerance is the induction of unresponsiveness in peripheral sites as a result of selfreactive lymphocytes encountering self-antigens under particular conditions. In B cells this results in apoptosis, or anergy which leads to eventual elimination from the B cell pool.

Mature B Cells

Bone Marrow Bone Marrow

IgM IgD IgM

Periphery
Y Y

Y Y Y Y Y Y

Antigen

Plasma Cell
Lymphoid ProStem Cell B Cell

PreB Cell

Immature Mature Activated B Cell B Cell B Cell Memory B Cell

Antigen Independent

Antigen Dependent
Secondary Lymphoid Organs

Primary Lymphoid Organ

CONCEPT: Co-expression of IgM and IgD results from alternative RNA splicing.