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Repertoire of the Mouse: An Analysis of the Usage of Two V Gene Segments1
Immunology Program, Sackler School of Graduate Biomedical Sciences, and the Department of Pathology, Tufts University School of Medicine, Boston, MA 02111
Detailed analysis of the rearrangement and expression of two mouse
V
genes has been used to examine B cell repertoire development. The
V1-A gene is used by a large proportion (9.6%) of splenic B cells
in the adult primary repertoire, whereas the V22 gene is used at a
much lower frequency (0.16%). Consistent with these results,
quantitative PCR (Q-PCR) assays revealed that the number of splenic B
cells with rearranged V1-A genes is much greater than the number
with rearranged V22 genes. Q-PCR was also performed on both normal
bone marrow pre-B cells and transformed pre-B cells induced to
rearrange their loci at high frequency. In contrast to splenic B
cell rearrangements, the numbers of V1-A and V22 rearrangements
in pre-B cells differ by only two- or threefold, suggesting that the
intrinsic rearrangement frequencies of these two V genes are not
significantly different. Further evidence of disproportionate selection
was obtained by comparing the percentages of productive rearrangements
amplified from genomic splenic DNA. Sequence analysis showed 84% (37
of 44) of the V1-A rearrangements but only 57% (29 of 51) of the
V22 rearrangements to be in-frame. Together these results suggest
that B cells expressing V1-A-encoded light chains are preferentially
selected either in the periphery or in the transition from pre-B to B
cell. Sequence data also reveal a surprisingly restricted diversity of
VJ junctions, apparently due to biases introduced by the rearrangement
mechanism.
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