内毒素耐受状态下胸腺细胞TCRβ链CDR3区多态性和长度的变化  

Change of polymorphism and length of TCRβ chain CDR3 region of thymus T cells in the condition of endotoxin-induced tolerance

作　　者：贾力[1,2] 陶弋婧 赵娟娟[1,2] 郭萌萌[1,2] 陈超[1,2] 褚风云 王海嵘[1,2] 刘云[2] 罗军敏[1,2] 姚新生[1,2] 徐林[1,2] 

机构地区：[1]遵义医学院免疫学教研室暨贵州省生物治疗人才基地,贵州遵义563099 [2]遵义医学院贵州省普通高等学校特色药物肿瘤防治特色重点实验室,贵州遵义563099

出　　处：《遵义医学院学报》2017年第2期139-142,149,共5页Journal of Zunyi Medical University

基　　金：贵州省高层次创新型人才计划项目(NO:黔科合人才[2016]4031);遵义医学院优秀青年人才计划项目(NO:15ZY-001)

摘　　要：目的研究内毒素耐受状态下小鼠胸腺T细胞TCRβ链CDR3多态性和长度的变化,并探讨其意义。方法以5mg/kg浓度LPS腹腔注射C57BL/6小鼠建立内毒素耐受模型,5 mg/kg浓度生理盐水腹腔注射72 h后为对照组,提取对照组、模型后72 h实验组和8 d实验组的小鼠胸腺组织RNA,并逆转录为c DNA;以22条TRBV基因家族序列设计上游引物,TRBJ1-1基因家族序列设计下游引物,PCR扩增出完整的TCRβ链CDR3区;毛细管电泳技术分析TCRβ链CDR3区的多态性和长度。结果正常小鼠胸腺样本多数呈现中间高两边低的正态性分布的TCRβ链CDR3谱型峰图,少数呈现偏峰、寡峰或低峰;LPS注射后72 h胸腺样本TCRβ链CDR3区呈现数量不等的偏峰、寡峰和低峰;模型8 d胸腺样本大多呈现多峰。此外,与正常组相比,模型72 h和8 d胸腺样本TCRβ链CDR3区产物长度均发生了不同程度的变化。结论内毒素耐受状态下,小鼠胸腺细胞TCRβ链CDR3区的多态性和长度均发生了明显改变,为后续进一步研究该状态下胸腺细胞发育及功能变化提供前期实验基础。Objective To observe the change of polymorphism and length of TCRβ chain CDR3 region of thymus T cells in murine endotoxin-induced tolerance and explore its significance.Methods Endotoxin-induced tolerance model in C57/BL6 mice was established by intraperitoneal injection of LPS （5 mg/kg）,while the normal mice were intraperitoneally injected with NS （5 mg/kg）.The total RNA of the thymus tissue was extracted at 72 hours and 8 days of post LPS injection and reverse transcribed into cDNA,resepectively.Upstream primer sequence was designed according to 22 TRBV gene families separately and TRBJ1-1 gene families sequence was used to design the downstream primer.PCR assay was used to amplify the complete TCRβ chain CDR3 region.The CDR3 length and diversity were analyzed by capillary electrophoresis.Results Most of the thymus samples of the normal mice presented the standard Gaussian distribution.But there were few partial-peak/oligo-peak or low-peak.Different number of partial-peak/oligo-peak or low-peak at 72 hours of post-LPS injection was shown.Eight days after the model establishment,most of the families presented poly-peak.In addition,compared with normal group,different degrees of changes on the length of TCRβchain CDR3 region of thymus T cells were indicated at 72 hours and 8 days of post LPS injection.Conclusion The polymorphism and length of TCRβ chain CDR3 region in thymus T cells are significant changed in the condition of endotoxin-induced tolerance,which provides the preliminary experimental basis for further study on the development and function of thymus cells in endotoxin-induced tolerance.

关 键 词：内毒素 胸腺 TCRβ链CDR3区 小鼠 

分 类 号：R392[医药卫生—免疫学]