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Paired TCRαβ analysis of virus‐specific CD8 + T cells exposes diversity in a previously defined ‘narrow’ repertoire
Author(s) -
Cukalac Tania,
Kan WanTing,
Dash Pradyot,
Guan Jing,
Quinn Kylie M,
Gras Stephanie,
Thomas Paul G,
La Gruta Nicole L
Publication year - 2015
Publication title -
immunology and cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0818-9641
DOI - 10.1038/icb.2015.44
Subject(s) - t cell receptor , repertoire , biology , epitope , cd8 , immune system , cytotoxic t cell , antigen , genetics , t cell , virology , physics , acoustics , in vitro
T‐cell receptor (TCR) usage has an important role in determining the outcome of CD8 + cytotoxic T‐lymphocyte responses to viruses and other pathogens. However, the characterization of TCR usage from which such conclusions are drawn is based on exclusive analysis of either the TCRα chain or, more commonly, the TCRβ chain. Here, we have used a multiplexed reverse transcription‐PCR protocol to analyse the CDR3 regions of both TCRα and β chains from single naive or immune epitope‐specific cells to provide a comprehensive picture of epitope‐specific TCR usage and selection into the immune response. Analysis of TCR repertoires specific for three influenza‐derived epitopes (D b NP 366 , D b PA 224 and D b PB1‐F2 62 ) showed preferential usage of particular TCRαβ proteins in the immune repertoire relative to the naive repertoire, in some cases, resulting in a complete shift in TRBV preference or CDR3 length, and restricted repertoire diversity. The NP 366 ‐specific TCRαβ repertoire, previously defined as clonally restricted based on TCRβ analysis, was similarly diverse as the PA 224 ‐ and PB1‐F2 62 ‐specific repertoires. Intriguingly, preferred TCR characteristics (variable gene usage, CDR3 length and junctional gene usage) appeared to be able to confer specificity either independently or in concert with one another, depending on the epitope specificity. These data have implications for established correlations between the nature of the TCR repertoire and response outcomes after infection, and suggest that analysis of a subset of cells or a single TCR chain does not accurately depict the nature of the antigen‐specific TCRαβ repertoire.