Early acquisition of cytolytic function and transcriptional changes in a primary CD8+ T-cell response in vivo
Early acquisition of cytolytic function and transcriptional changes in a primary CD8+ T-cell response in vivo
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DOI：
10.1182/blood-2006-03-011643
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发表时间：
2007-02-01
期刊：
影响因子：
20.3
通讯作者：
Callan, Margaret F. C.
中科院分区：
文献类型：
--
作者：
Chiu, Christopher;Heaps, Adrian G.;Callan, Margaret F. C.
功能研究表明，CD 8（+）T细胞的编程发生在最初抗原接触后的2小时内。为了确定这些事件的分子基础，我们将来自F5 Rag（-/-）小鼠的TCR转基因T细胞转移到幼稚受体中，并用表达免疫显性流感表位NP 366374的重组牛痘刺激它们。使用Affyphase 430 2.0基因芯片和定量聚合酶链反应（PCR）分析表位特异性细胞毒性T淋巴细胞（CTL）中的转录。我们证明了一个早期的转录爆发与最大数量的基因达到峰值表达刺激后12小时。使用体内细胞毒性测定，我们证明了溶细胞基因的早期上调伴随着在刺激24小时内获得杀伤能力。然而，直到48小时才观察到T细胞增殖。因此，我们得出结论，克隆扩增，而不是收购效应功能是限速步骤的发展中的主要CTL反应。
Functional studies show that programming of CD8(+) T cells occurs early after initial antigen encounter within as little as 2 hours. To define the molecular basis of these events, we transferred TCR transgenic T cells from F5 Rag(-/-) mice into naive recipients and stimulated them with recombinant vaccinia expressing the immumodominant influenza epitope NP366374. Transcription in epitope-specific cytotoxic T lymphocytes (CTLs) was analyzed using Affymetrix 430 2.0 Gene-Chips and quantitative polymerase chain reaction (PCR). We demonstrated an early transcriptional burst with the greatest number of genes reaching peak expression 12 hours after stimulation. Using in vivo cytotoxicity assays we demonstrated that early up-regulation of cytolytic genes was accompanied by acquisition of killing capacity within 24 hours of stimulation. However, T-cell proliferation was not observed until 48 hours. We therefore conclude that clonal expansion rather than acquisition of effector function is the rate-limiting step in the development of a primary CTL response.