| First Author | Aoki-Ota M | Year | 2012 |
| Journal | J Immunol | Volume | 188 |
| Issue | 5 | Pages | 2305-15 |
| PubMed ID | 22287713 | Mgi Jnum | J:181249 |
| Mgi Id | MGI:5310659 | Doi | 10.4049/jimmunol.1103484 |
| Citation | Aoki-Ota M, et al. (2012) Skewed primary igkappa repertoire and v-j joining in C57BL/6 mice: implications for recombination accessibility and receptor editing. J Immunol 188(5):2305-15 |
| abstractText | Previous estimates of the diversity of the mouse Ab repertoire have been based on fragmentary data as a result of many technical limitations, in particular, the many samples necessary to provide adequate coverage. In this study, we used 5'-coding end amplification of Igkappa mRNAs from bone marrow, splenic, and lymph node B cells of C57BL/6 mice combined with amplicon pyrosequencing to assess the functional and nonfunctional Vkappa repertoire. To evaluate the potential effects of receptor editing, we also compared V/J associations and usage in bone marrows of mouse mutants under constitutive negative selection or an altered ability to undergo secondary recombination. To focus on preimmune B cells, our cell sorting strategy excluded memory B cells and plasma cells. Analysis of approximately 90 Mbp, representing >250,000 individual transcripts from 59 mice, revealed that 101 distinct functional Vkappa genes are used but at frequencies ranging from approximately 0.001 to approximately 10%. Usage of seven Vkappa genes made up >40% of the repertoire. A small class of transcripts from apparently nonfunctional Vkappa genes was found, as were occasional transcripts from several apparently functional genes that carry aberrant recombination signals. Of 404 potential V-J combinations (101 Vkappas x 4 Jkappas), 398 (98.5%) were found at least once in our sample. For most Vkappa transcripts, all Jkappas were used, but V-J association biases were common. Usage patterns were remarkably stable in different selective conditions. Overall, the primary kappa repertoire is highly skewed by preferred rearrangements, limiting Ab diversity, but potentially facilitating receptor editing. |
