Lehner P J, Wang E C, Moss P A, Williams S, Platt K, Friedman S M, Bell J I, Borysiewicz L K
Department of Medicine, University of Wales College of Medicine, Heath Park, Cardiff.
J Exp Med. 1995 Jan 1;181(1):79-91. doi: 10.1084/jem.181.1.79.
The major histocompatibility complex class I-restricted cytotoxic T lymphocyte (CTL) response is important in the clearance of viral infections in humans. After influenza A infection, a peptide from the matrix protein, M58-66, is presented in the context of the MHC allele HLA-A0201 and the resulting CTL response is detectable in most HLA-A0201 subjects. An initial study suggested that M58-66-specific CTL clones show conserved T cell receptor (TCR) alpha and beta gene segments. We have addressed the significance of this observation by determining the expression of V beta 17 during the development of M58-66-specific CTL lines in 21 unrelated HLA-A0201 subjects, and analyzing TCR usage by M58-66-specific CTL clones. TCR V beta 17 was the dominant V beta segment used and CD8 V beta 17 expansion correlated with M58-66-specific lysis. Limiting dilution analysis from five subjects showed the M58-66 CTL precursor frequency to vary between 1/54,000 and less than 1/250,000, and that up to 85% of the matrix peptide (M58-66)-specific CTL used the V beta 17 gene segment. The M58-66 specific CTL response was dependent on previous viral exposure and specific V beta 17 expansion, as it was not found in cord blood, despite a readily expandable V beta 17+ CD8+ T cell subpopulation. Sequence analysis of 38 M58-66-specific V beta 17 transcripts from 13 subjects revealed extensive conservation in the CDR3 region including conservation of an arginine-serine motif. To test the dependence of this CTL response on the V beta 17 gene segment, peripheral blood lymphocytes were depleted of CD8+ TCR V beta 17+ cells, before the generation of M58-66-specific CTL. In most cases such depletion blocked or severely reduced the generation of the M58-66-specific response, and under limiting dilution conditions could abolish M58-66-specific CTL precursors. These studies reveal the dependence of this natural human immune response on a particular TCR gene segment.
主要组织相容性复合体I类限制性细胞毒性T淋巴细胞（CTL）反应在人类清除病毒感染中起着重要作用。甲型流感病毒感染后，基质蛋白的一段肽M58 - 66在MHC等位基因HLA - A0201的背景下呈递，并且在大多数HLA - A0201个体中可检测到由此产生的CTL反应。一项初步研究表明，M58 - 66特异性CTL克隆显示出保守的T细胞受体（TCR）α和β基因片段。我们通过测定21名不相关的HLA - A0201个体中M58 - 66特异性CTL系发育过程中Vβ17的表达，并分析M58 - 66特异性CTL克隆的TCR使用情况，来探讨这一观察结果的意义。TCR Vβ17是所使用的主要Vβ片段，CD8 Vβ17的扩增与M58 - 66特异性裂解相关。对5名个体的有限稀释分析表明，M58 - 66 CTL前体频率在1/54,000至小于1/250,000之间变化，并且高达85%的基质肽（M58 - 66）特异性CTL使用Vβ17基因片段。M58 - 66特异性CTL反应依赖于先前的病毒暴露和特定的Vβ17扩增，因为尽管存在易于扩增的Vβ17 + CD8 + T细胞亚群，但在脐血中未发现该反应。对13名个体的38条M58 - 66特异性Vβ17转录本的序列分析显示，互补决定区3（CDR3）区域存在广泛的保守性，包括精氨酸 - 丝氨酸基序的保守性。为了测试这种CTL反应对Vβ17基因片段的依赖性，在产生M58 - 66特异性CTL之前，去除外周血淋巴细胞中的CD8 + TCR Vβ17 + 细胞。在大多数情况下，这种去除会阻断或严重减少M58 - 66特异性反应的产生，并且在有限稀释条件下可以消除M58 - 66特异性CTL前体。这些研究揭示了这种天然人类免疫反应对特定TCR基因片段的依赖性。
