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B- and T-cell receptor genes

Dept of Cell and Molecular Biology!

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Immunoglobulin Variable regions

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RNA-DNA heteroduplex analysis of an immunoglobulin light chain gene

Mouse H and L chain loci!

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Variability generating mechanisms in immunoglobulin and TCR genes!

! ! ! Germline diversity; a large number of different V, D and J segments which differ in nucleotide sequence. Combinatorial diversity; the different V, D, and J segments can freely combine, resulting in a large number of possible combinations(VxDxJ). Junctional diversity; variation in the position of the cleavage site at the junctions between V and J segments in the light chain and the V and D and D and J in the heavy chain (approximately +3 to -3 nt). P-region diversity; difference in where the ligated loops are cleaved generates additional diversity at the V-J, V-D and D-J joining sites. N-region divesity; addition or removal of up to approximately 10 bp at either side of the D segment in the IgH and TCR! and " genes, and at a lesser extent at V- J junctions. Generated by the enzyme TdT (Terminal doeoxy nuclotide Transferase). Somatic mutations; random exchange of single nucleotides (one at a time) in and around the rearranged V region. Only found in the Ig genes and not in the TCR genes. Free combination of light and heavy chain polypeptides (TCR # and ! or TCR $ and " polypeptides)

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Variability in immunoglobulin and TCR genes!

! Total potential diversity with junctional diversity; Immunoglobulins 10 11. TCR #! 10 16 TCR $" 10 18 ! Total potential diversity with somatic mutations ; Immunoglobulins 1015-16

Transcriptional activation of the %-light chain locus!

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Transcriptional activation of the heavy and light chain locus!

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Differences between T cell recptors and immunoglobulins during somatic rearrangements !

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Switch Regions
G rich, simple repetitive sequences 1-10 kb in length. IgE (&) switch region approx. 1kb. IgG1 ($1) switch region approx 10 kb. Short 5-7 nt repeats like, GAGCT, GGGGT, GGGCT S , S & and S # are closely related with 20, 40 and 80 bp repeat units respectively. S $ the most divergent with 49-52 bp repeat units

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Transcriptional activation of un-rearranged locus

I-exon Promoter
AAAAAA

Switch region (S&)

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Germline Transcripts (GL) are essential for switch recombination. Activated by IL-4 and CD40-CD40L interaction. GL transcripts stimulate recombination not only by opening of chromatin. The specific transcript or splicing is of importance, 10-100 fold higher switch frequency with the I exon (than irrelevant transcription)

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Membrane and secretory forms

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B cell development Early events!

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Surface markers on peripheral B cells

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