Comprehensive analysis of the functional TCR repertoire at the single-cell level

被引：28

作者：

Ozawa, Tatsuhiko ^{[1
]}

Tajiri, Kazuto ^{[1
]}

Kishi, Hiroyuki ^{[1
]}

Muraguchi, Atsushi ^{[1
]}

机构：

[1] Toyama Univ, Grad Sch Med & Pharmaceut Sci, Dept Immunol, Toyama 9300194, Japan

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2008年 / 367卷 / 04期

关键词：

comprehensive analysis; single-cell analysis; TCR repertoire; single-cell 5 '-RACE; superantigen; staphylococcus enterotoxin B (SEB);

D O I：

10.1016/j.bbrc.2008.01.011

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A V beta TCR repertoire is analyzed for understanding the T-cell population in the immune response. However, the TCR repertoire of the V alpha-V beta pair is difficult to analyze because no suitable analytical method is available. Here, we have applied the single-cell 5'-RACE method for amplifying TCR cDNAs from single T-cells and analyzed the repertoire of V alpha-V beta pairs in human T-cells that responded to a superantigen, SEB. We found that the TCR V beta profile of the SEB-stimulated CD4(+) T-cells was in accordance with the previous reports, that the TCR V alpha profile also exhibited a prominent difference, and that the TCR V alpha-V beta pairs of the SEB-responding T-cells were promiscuous. We have also found a significant dual TCR alpha expression in single T-cells. This is the first report of a comprehensive analysis of the functional repertoire of V alpha-V beta pairs at the single T-cell level. This novel method may contribute to TCR-based immunotherapeutics. (c) 2008 Elsevier Inc. All rights reserved.

引用

页码：820 / 825

页数：6

共 28 条

[1] A quantitative method of determining initial amounts of DNA by polymerase chain reaction cycle titration using digital imaging and a novel DNA stain [J].