Knockdown of T-bet expression in Mart-127-35 -specific T-cell-receptor-engineered human CD4(+) CD25(-) and CD8(+) T cells attenuates effector function.
Knockdown of T-bet expression in Mart-127-35 -specific T-cell-receptor-engineered human CD4(+) CD25(-) and CD8(+) T cells attenuates effector function.
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Mart-127-35 特异性 T 细胞受体工程化人类 CD4( )、CD25(-) 和 CD8( ) T 细胞中 T-bet 表达的敲低会减弱效应子功能。
DOI：
10.1111/imm.12431
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发表时间：
2015
期刊：
影响因子：
6.4
通讯作者：
Dorsky,DavidI
中科院分区：
文献类型：
--
作者：
Jha,SidharthS;Chakraborty,NityaG;Singh,Prashant;Mukherji,Bijay;Dorsky,DavidI
基因转移创造肿瘤表位特异性细胞溶解T细胞用于癌症的过继免疫治疗仍然是一个积极探索的领域。当Mart - 127-35特异性DMF5 T细胞受体（TCR）被转移到外周人CD4+T细胞中时，重编程细胞表现出T辅助型1 （Th1）表型，具有显著的多因子效应能力。T - bet转录因子在决定Th1分化途径中起着重要作用。为了更深入地了解T - bet如何通过基因转移控制人类T细胞重编程的结果，我们开发了一个系统，用于检测短发夹RNA介导的T - bet基因敲低在均匀表达敲低结构的分类细胞群体中的影响。在这个系统中，使用活化的外周人CD4+CD25 -和CD8+T细胞，T - bet敲低导致干扰素γ对抗原特异性和非特异性TCR刺激的反应衰减。白细胞介素- 2 （IL - 2）抗原特异性反应未被T - bet敲除而减弱。此外，在TCR重编程的CD8+细胞中，T - bet敲低可减弱细胞溶解效应反应。T - bet敲除不会导致重定向进入Th2分化途径，并且在该系统中未检测到IL - 4、IL - 10或IL - 17反应增加。这些结果表明，在TCR重编程的人T细胞中，T - bet表达是维持CD4+CD25 -和CD8+效应表型所必需的。他们还表明，逆转录载体和慢载体转导所必需的激活方案可能会使重编程细胞进入Th1表型，这种表型不能通过T - bet敲除而改变，但是，干扰素γ基因激活持续需要T - bet表达。
Gene transfer to create tumour epitope‐specific cytolytic T cells for adoptive immunotherapy of cancer remains an area of active inquiry. When the Mart‐127–35‐specific DMF5 T‐cell receptor (TCR) is transferred into peripheral human CD4+T cells, the reprogrammed cells exhibit a T helper type 1 (Th1) phenotype with significant multifactorial effector capabilities. The T‐bet transcription factor plays an important role in determination of the Th1 differentiation pathway. To gain a deeper understanding of how T‐bet controls the outcome of human T‐cell reprogramming by gene transfer, we developed a system for examining the effects of short hairpin RNA‐mediated T‐bet gene knockdown in sorted cell populations uniformly expressing the knockdown construct. In this system, using activated peripheral human CD4+CD25−and CD8+T cells, T‐bet knockdown led to attenuation of the interferon‐γresponse to both antigen‐specific and non‐specific TCR stimulation. The interleukin‐2 (IL‐2) antigen‐specific response was not attenuated by T‐bet knockdown. Also, in TCR‐reprogrammed CD8+cells, the cytolytic effector response was attenuated by T‐bet knockdown. T‐bet knockdown did not cause redirection into a Th2 differentiation pathway, and no increased IL‐4, IL‐10, or IL‐17 response was detected in this system. These results indicate that T‐bet expression is required for maintenance of the CD4+CD25−and CD8+effector phenotypes in TCR‐reprogrammed human T cells. They also suggest that the activation protocol necessary for transduction with retrovectors and lentivectors may commit the reprogrammed cells to the Th1 phenotype, which cannot be altered by T‐bet knockdown but that there is, nevertheless, a continuous requirement of T‐bet expression for interferon‐γgene activation.