T淋巴瘤EL-4细胞双受体表达研究  

Expression of two distinct T cell receptors by EL-4 T lymphoma cells

作　　者：张媛[1] 罗微[1] 马骊[1] 

机构地区：[1]南方医科大学生物技术学院分子免疫学研究所,广州510515

出　　处：《中国免疫学杂志》2011年第5期404-408,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(30972680;30771952);教育部新世纪优秀人才支持计划(NCET-07-0410)资助

摘　　要：目的:检测T淋巴瘤EL-4细胞T细胞受体(T cell receptor,TCR)的表达情况,为进一步研究T细胞等位基因排斥机制奠定基础。方法:以小鼠脾细胞作为阳性对照,分别提取脾细胞和EL-4细胞mRNA,逆转录为cDNA,RT-PCR扩增24个TCR BV家族CDR3区,结合基因扫描(GeneScan)和基因测序技术,对有表达的TCR BV家族进行CDR3谱型和序列分析。结果:小鼠脾细胞24 BV家族均有表达,各TCR BV家族CDR3谱型均呈高斯分布(Gaussian distribution),表明24 BV家族均为多克隆性。EL-4细胞被同时检测到TCR BV10和BV12家族表达,CDR3谱型均呈单峰,两个家族的RT-PCR产物经测序鉴定证实确属TCR序列,且均为框内编码。结论:EL-4细胞存在两套框内重排的TCRβ链,表明其TCRβ链可能存在等位基因排斥"缺陷"现象。本研究为探索T细胞等位基因排斥机制奠定了基础。Objective：To investigate the expression of T cell receptor（TCR） inEL-4 T lymphoma cells.Methods：Mouse spleen cells were used as positive control,total RNA was isolated from mouse spleen cells and EL-4 cell line.Complementarity Determining Region 3（CDR3） of 24 TCR BV gene subfamilies was amplified by the reverse transcription polymerase chain reaction（RT-PCR）.To determine the spectratypes and sequences of CDR3 region of TCR BV subfamilies expressed by EL-4,PCR products were analyzed by TCR GeneScan Technique and DNA sequencer,respectively.Results：24 TCR BV subfamilies of mouse spleen cells were polyclonal,the spectratypes of TCR CDR3 region were showed Gaussian distribution.EL-4 was shown detected the expression of TCR BV10 and BV12 simultaneously,spectratypes of which showed single peak,and the TCR sequences of BV10 and BV12 were confirmed by DNA sequencer and they were both in-frame rearrangements.Conclusion：Two sets of in-frame rearrangements of TCR β chain exist in EL-4,and there maybe ＇defects＇ in allelic exclusion in TCR β chain.It provides a basis for future research of allelic exclusion mechanism.

关 键 词：EL-4细胞株 T细胞受体(TCR) 等位基因排斥“缺陷” 框内重排 

分 类 号：R392.12[医药卫生—免疫学]