The genes for perforin, granzymes A-C and IFN-γ are differentially expressed in single CD8+ T cells during primary activation
The genes for perforin, granzymes A-C and IFN-γ are differentially expressed in single CD8+ T cells during primary activation
复制标题
DOI：
10.1093/intimm/dxf028
复制
发表时间：
2002-06-01
影响因子：
4.4
通讯作者：
Fitzpatrick, DR
中科院分区：
文献类型：
--
作者：
Kelso, A;Costelloe, EO;Fitzpatrick, DR
在这里，我们发现穿孔素、三种主要的T细胞颗粒酶(A-C)和干扰素-γ基因在初始CD8(+)T细胞的初始激活过程中存在动态和单细胞水平的差异表达。当CD44(低)CD62L(高)CD8(+)T细胞被IL-2激活并固定CD3、CD8和CD11a抗体时，穿孔素、颗粒酶B和干扰素-γmRNAs的表达在第2天就被诱导，并与穿孔素依赖的细胞溶解活性平行增加。颗粒酶C和A的转录本分别在1天和3天后才被检测到。单细胞聚合酶链式反应显示，每种基因的表达频率与总水平呈平行上升，但单个细胞表达穿孔素、颗粒酶A-C和干扰素-γmRNAs的不同组合。这些表达模式表明，颗粒酶A和C的延迟表达不是由于不同细胞亚群的延迟激活所致。对数据的统计分析表明，每个基因在单细胞水平上存在差异调控。单个原始CD8(+)T细胞在7天后形成克隆，在克隆水平上表达所有五种产物，但也在单细胞水平上表达不同的组合。我们的结论是，在初级激活过程中，CD8(+)T细胞逐渐获得了表达大部分或全部这些基因的能力，并且克隆和群体内单个细胞之间观察到的可变表达模式反映了表达谱的瞬时差异，而不是可遗传的差异。
Here we show that the genes for perforin, the three major T cell granzymes (A-C) and IFN-gamma are differentially expressed during primary activation of naive CD8(+) T cells, kinetically and at the single-cell level. When CD44(low)CD62L(high)CD8(+) lymph node T cells were activated with IL-2 and immobilized antibodies to CD3, CD8 and CD11a, expression of perforin, granzyme B and IFN-gamma mRNAs was induced by day 2, and increased in parallel with perforin-dependent cytolytic activity. Granzyme C and A transcripts were not detected until 1 and 3 days later respectively. Single-cell PCR showed that expression frequencies rose in parallel with total levels of each mRNA, but that individual cells expressed diverse combinations of perforin, granzyme A-C and IFN-gamma mRNAs. These expression patterns indicated that the delayed expression of granzymes A and C was not due to late activation of distinct cell subpopulations. Statistical analysis of the data suggested that each gene was differentially regulated at the single-cell level. Individual naive CD8(+) T cells gave rise over 7 days to clones that expressed all five products at the clonal level, but also expressed diverse combinations at the single-cell level. We conclude that, during primary activation, CD8(+) T cells progressively acquired the ability to express most or all of these genes, and that the variable expression patterns observed among single cells within clones and populations reflected transient rather than heritable differences in expression profile.