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作 者:邹红云[1] 马骊[1] 姚新生[1] 温茜[1] 罗微[1] 王小宁[2]
机构地区:[1]南方医科大学生物技术学院分子免疫研究所,广东广州510515 [2]华南理工大学生物科学与工程学院,广东广州510641
出 处:《南方医科大学学报》2006年第7期939-943,共5页Journal of Southern Medical University
基 金:国家重点基础研究发展规划"973"项目(2001CB510008)~~
摘 要:目的探讨Jurkat细胞TCRBD2-BJ2基因重排对TCRβ链可变区互补决定区3(BVCDR3)可能产生的影响,为深入研究TCR基因重排奠定实验基础。方法RT-PCR扩增TCRBV26个亚家族CDR3区,结合TCR基因扫描(GeneScan)和基因测序技术,监测Jurkat细胞在增殖传代过程中以及在T细胞激活剂和超抗原SEA等刺激诱导后TCRBVCDR3谱系漂移及长度和序列变化。结果表达TCRBV8家族的单克隆细胞株Jurkat,在增殖传代过程中,以及经刺激诱导48或72h后,未监测到有TCRBV8以外的新的BV亚家族出现,BV8CDR3长度和一级核苷酸序列也未发生变化。结论Jurkat细胞发生的TCR基因重排可能不引起TCRBVCDR3改变,因而对TCRBVCDR3区抗原识别特异性(即抗原特异性漂移)并未产生影响,但并不排除TCR发生改变的可能性。Objective To investigate the effects of T cell receptor (TCR) BD2-BJ2 gene rearrangement on the complementary-determining region (CDR) 3 of TCR β chain (TCR BV CDR3) in Jurkat cells. Methods TCR BV gene subfamilies were detected by RT-PCR in Jurkat cells during proliferation and after induction with non-specific T cell activators and SEA, respectively. To determine the clonality of TCR BV subfamilies and the lengths of CDR3, the PCR products were analyzed by TCR GeneScan technique, and the sequences of CDR3 were further analyzed by DNA sequencer. Results No new TCR BV subfamilies were found in Jurkat cells, a monoclonal BV8^+ cell line, either during cell proliferation or after stimulation with different treatments, nor were any differences found in CDR3 size or sequences. Conclusions TCR BD2-BJ2 rearrangement in Jurkat cells may not play a role in modification of TCR BV CDR3 domains or the consequent antigen immunorecognition ofBV CDR3, but the possibility of TCR modification can not be excluded.
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