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Asymmetric Contribution to Ig Repertoire Diversity by V Exons: Differences in the Utilization of V10 Exons

Center for Biologics Evaluation and Research, Food and Drug Administration, Rockville, MD 20892

The mouse has approximately 140 germline V genes, and functional V exons are expressed at roughly equivalent levels in the preimmune repertoire. We have examined the expression of individual members of the V10 family. V10A and V10B genes have been utilized in numerous hybridomas and myelomas, while V10C has not. In this study, we have cloned the V10C gene and shown that it is structurally functional, has the expected promoter elements and recombination signal sequences, and that it is capable of recombination. V10C mRNA, however, is present at levels at least 1000-fold lower than V10A and V10B in adult spleens. While there are no sequence differences in the octamer or TATA box between V10C and V10A, there are three nucleotide changes in the promoter region. These promoters equally drive the expression of a reporter gene in B cells or plasma cells, but the V10A promoter is able to drive expression in pre-B cell lines significantly better than the V10C promoter (p < 0.05). V10C rearrangements can be detected in bone marrow and splenic DNA. Therefore, the lack of V10C expression may reflect the inability of V10C-rearranged cells to undergo positive or negative selection. Our results suggest that the available Ab repertoire is shaped not only by the number of structurally functional genes, but also by the ability of assembled genes to be expressed at critical points during B cell maturation.

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