B Cell Development and Primary Antibody (Ab) Repertoire Flashcards
Describe the three gene families and how they can be divided.
Each of the 3 gene families, the kappa light chain family, the lambda light chain family and the heavy chain family can be divided into V-region genes and C-region genes.
The kappa, lambda and H chains are located on separate chromosomes. Each set of genes, kappa, lambda and heavy chain, has a similar basic organization.
The Ig heavy and light chain loci are composed of multiple genes that give rise to the V and C regions of the proteins, separated by stretches of non-coding DNA.
Where are V region exons found?
At the 5’end of each Ig locus are the V region exons each about 300 base-pairs (bp) long, separated from one another by non-coding DNA of varying lengths
Where are joining (J) and diversity (D) segments found?
What do they code for?
Downstream of the V genes are additional coding sequences, 30 to 50 bp long, which make up the joining (J) segments and, in the H chain locus only, the diversity (D) segments
The J and D gene segments code for the carboxy terminal ends of the V regions, including the third hypervariable (complementarily-determining) regions of antibody molecules
What encodes the variable and constant regions in light chain and heavy chain proteins?
Thus, in an Ig light chain protein (kappa or lambda), the variable region is encoded by the V and J exons and the constant region by a C exon. In the heavy chain protein, the variable region is encoded by the V, D, and J exons. The constant region of the protein is derived from the multiple C exons and, for membrane-associated heavy chains, the exons encoding the transmembrane and cytoplasmic domains.
Describe the C region genes.
Where are they? How are they arranged?
What does eaach heavy chain C region gene consist of? (How many exons?)
At varying distances 3’ of the V genes are the C region genes. In both mouse and man, the kappa light chain locus and a single C-kappa gene and the genes for heavy chain C regions (CH) of different isotypes are arranged in a tandem array.
Each heavy chain C region gene actually consists of three to four exons (each similar in size to a V region exon) that make up up the complete C region, and smaller exons that code for the carboxy terminal transmembrane (TM) and cytoplasmic domains of the heavy chains.
Describe how Ig genes are expressed in B-lineage cells and non-B-lineage cells.
All cells except B-lineage, including plasma cells contain Ig genes in the germline configuration. The Ig genes are expressed only in B-lineage cells.
What is the first step in production of antibodies?
Rearrangements of Ig genes are the essential first steps in the production of antibodies.
Describe how DNA rearrangements occur in regards to antigen stimulation.
DNA rearrangements occur in a precise order and occur independent of antigen stimulation.
What is the first Ig gene rearrangement in the heavy chain?
First step to make immature B lymphocyte
Heavy chain - DJ. The first Ig gene rearrangement involves the heavy chain locus and leads to joining of one D and one J gene segment with deletion of the intervening DNA.
What follows the DJ rearrangement in the heavy chain?
What is important to note about this rearrangement?
(2nd step to make immature B lymphocyte)
Following the DJ rearrangement, one of the many V genes is joined to the DJ complex, giving rise to a rearranged VDJ gene. At this stage, all D segments 5’ of the rearranged D are also deleted.
This VDJ recombination occurs only in cells committed to become B lymphocytes and is a critical control point in Ig expression because only the rearranged V gene is subsequently transcribed. The C region genes remain separated from this VDJ complex by an intron.
Describe the rearrangement following VDJ.
3rd step to make immature B lymphocyte
Light chain - VJ. The next somatic DNA recombination involves a light chain locus. One V segment is joined to one J segment, forming a VJ complex, which remains separated from the C region by an intron, and this gives rise to the primary RNA transcript. Splicing of the intron from the primary transcript joins the C gene to the VJ complex, forming an mRNA that is translated to produce the κ protein. The light chain assembles with the previously synthesized μ to form the complete membrane IgM molecule, which is expressed on the cell surface, and the cell is now the immature B lymphocyte.
What does a single B cell make (major rule of antibody specificity).
What does a single plasma cell make?
A single B cell makes only one kind of antibody specificity (one VH and one VL), i.e., allelic exclusion occurs
Also, a single plasma cell makes only one kind of antibody; i.e., 1 kind of H chain & 1 kind of L chain; B cells may violate this rule & synthesize two or more heavy chain isotypes simultaneously for the cell surface, eg. IgM and IgD
What are ProB cells.?
These cells are precursors of PreB cells. They have IgH DJ gene rearrangements and no light chain gene rearrangements.
From what do all B lymphocytes arise?
Describe what the earliest cell type that synthesizes a detectable Ig gene product contains.
Where are pre-B lymphocytes found?
What is the pre-B receptor composed of?
All B lymphocytes arise in the bone marrow from a stem cell that does not produce Ig. The earliest cell type that synthesizes a detectable Ig gene product contains cytoplasmic mu-heavy chains composed of variable (V) and constant (C) regions. This cell is called the pre-B lymphocyte and is found only in hematopoietic tissues, such as the bone marrow and fetal liver. The pre-B receptor is comprised of surrogate light chain, mu-chain, Ig-alpha and Ig-beta
Describe immature B cells.
What happens at the next identifiable stage in B cell maturation?
What functions as specific receptors for antigens?
What are immature B lymphocytes? Why are they called this?
At the next identifiable stage in B cell maturation, kappa or lambda light chains are also produced.
These associate with mu heavy chains and then the assembled IgM molecules are expressed on the cell surface, where they function as specific receptors for antigens.
IgM-bearing B cells that are recently derived from bone marrow precursors are called immature B lymphocytes because they do not proliferate and differentiate in response to antigens. Once a B cell expresses a complete heavy or light chain, it cannot produce another heavy or light chain containing a different V region.