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PLoS One. 2018 Jan 10;13(1):e0190982. doi: 10.1371/journal.pone.0190982. eCollection 2018.

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

Author information

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 of America.

Abstract

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.

PMID:
29320559
PMCID:
PMC5761896
DOI:
10.1371/journal.pone.0190982
[Indexed for MEDLINE]
Free PMC Article

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