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Characterization of the naive murine antibody repertoire using unamplified high-throughput sequencing

Journal Article · · PLoS ONE
 [1];  [1];  [1];  [2];  [1];
  1. Division of Biology, Kansas State University, Manhattan, Kansas (United States of America).
  2. Division of Biomedical Engineering Sciences, Loma Linda University, Loma Linda, California (United States)
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Antibody specificity and diversity are generated through the enzymatic splicing of genomic gene segments within each B cell. Antibodies are heterodimers of heavy- and light-chains encoded on separate loci. We studied the antibody repertoire from pooled, splenic tissue of unimmunized, adult female C57BL/6J mice, using high-throughput sequencing (HTS) without amplification of antibody transcripts. We recovered over 90,000 heavy-chain and over 135,000 light-chain immunoglobulin sequences. Individual V-, D-, and J-gene segment usage was uniform among the three mouse pools, particularly in highly abundant gene segments, with low frequency V-gene segments not being detected in all pools. Despite the similar usage of individual gene segments, the repertoire of individual B-cell CDR3 amino acid sequences in each mouse pool was highly varied, affirming the combinatorial diversity in the B-cell pool that has been previously demonstrated. There also was some skewing in the V-gene segments that were detected depending on chromosomal location. This study presents a unique, non-primer biased glimpse of the conventionally housed, unimmunized antibody repertoire of the C57BL6/J mouse.

Sponsoring Organization:
USDOE Office of Science (SC)
DOE Contract Number:
AC02-06CH11357
OSTI ID:
1627850
Journal Information:
PLoS ONE, Vol. 13, Issue 1; ISSN 1932-6203
Publisher:
Public Library of Science
Country of Publication:
United States
Language:
English

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