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J Immunol. 2014 Apr 15;192(8):3637-44. doi: 10.4049/jimmunol.1303334. Epub 2014 Mar 12.

Single-cell and deep sequencing of IgG-switched macaque B cells reveal a diverse Ig repertoire following immunization.

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1
Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, SE-171 77 Stockholm, Sweden;

Abstract

The nonhuman primate model is important for preclinical evaluation of prophylactic and therapeutic intervention strategies. The recent description of the rhesus macaque germline Ig loci and establishment of a database of germline gene segments offer improved opportunities to delineate Ig gene usage in the overall B cell repertoire as well as in response to vaccination. We applied 454-pyrosequencing and single-cell RT-PCR of bulk and sorted memory B cells, respectively, to investigate IGHV gene segment expression in rhesus macaques. The two methods gave remarkably concordant results and identified groups of gene segments that are frequently or rarely used. We further examined the VH repertoire of Ag-specific memory B cells induced by immunization with recombinant HIV-1 envelope glycoproteins, an important vaccine component. We demonstrate that HIV-1 envelope glycoprotein immunization activates a highly polyclonal response composed of most of the expressed VH gene segments, illustrating the considerable genetic diversity of responding B cells following vaccination.

PMID:
24623130
PMCID:
PMC3993955
DOI:
10.4049/jimmunol.1303334
[Indexed for MEDLINE]
Free PMC Article

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