EV71疫苗诱导小鼠CD4^+ T细胞受体Vβ17基因家族克隆化改变  

The clonality of murine CD4^+ TCR Vβ17 gene family induced by EV71 vaccine

作　　者：高帆[1] 邵杰[1,2] 王一平[1,3] 李玫颖[1,4] 毛群颖[1] 姚昕[1] 邵强[2] 李凤祥[1] 梁争论[1] 王军志[1] 

机构地区：[1]中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室,北京100050 [2]河南师范大学生命科学院,河南新乡453007 [3]沈阳药科大学药学院,辽宁沈阳110016 [4]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016

出　　处：《中国病毒病杂志》2012年第5期360-365,共6页Chinese Journal of Viral Diseases

基　　金：国家科技支撑项目(2008BAI69B01);国家"十一五"艾滋病和病毒性肝炎等重大传染病防治科技重大专项(2009ZX10004-804)

摘　　要：目的分析EV71疫苗诱导BALB/c小鼠CD4+TCR Vβ基因家族克隆化改变,探讨EV71疫苗免疫机制。方法采用TCR克隆化检测试剂盒和基因扫描技术对EV71疫苗诱导的BALB/c小鼠CD4+TCRVβ基因家族克隆化进行分析,并结合ELISPOT方法、Luminex技术和体外微量中和试验研究EV71疫苗诱导的细胞免疫和体液免疫应答情况,探讨EV71疫苗免疫机制。结果通过对小鼠CD4+TCR Vβ1-20克隆化检测,发现免疫组的Vβ17基因家族出现单克隆或寡克隆改变,对照组Vβ17为多克隆化,未见克隆化改变。免疫组小鼠脾MNC(去除CD8+)IFN-γ和IL-6的分泌水平较对照组显著增高(P均<0.01),血清EV71中和抗体应答均为阳性。进一步测序发现,免疫组Vβ17的CDR3区氨基酸序列为TASQNTLY。结论成功筛选出EV71疫苗诱导BALB/c小鼠CD4+TCR特异性改变的Vβ基因家族为Vβ17,与MHC-EV71抗原肽特异性结合的CDR3区氨基酸序列为TASQNTLY。Objective To vaccinate mouse with enterovirus 71（EV71） vaccine and study the vaccine-induced clonality of the murine CD4＋ TCR Vβ gene family and further investigate the immune mechanism of this vaccine.Methods The clonality of the murine CD4＋ TCR Vβ gene family was detected by TCRExpressTM Mouse T Cell Receptor Vβ Repertoire Clonality Detecting Kit and the genetic codes of CDR3 area were tested.ELISPOT assay,Luminex assay and neutralization test were used to study the cellular and humoral immune responses induced by EV71 vaccine.Results The clonality of the murine CD4＋ TCR Vβ1-20 family was detected and among which,Vβ1-17 gene exhibited monoclonality and oligoclonality in the immunized mice,while only polyclonality was detected in control mice.CD8＋-depleted spleen mononuclear cells from the immunized mice secreted much higher levels of IFN-γ and IL-6 than that from control mice（P〈0.01）.All mice in the immunized group showed positive EV71 antibody conversion and sequencing analysis identified that the CDR3 amino sequence of Vβ17 gene in the immunized group was TASQNTLY.Conclusions BALB/c mouse CD4＋TCR Vβ1-17 gene family was screened out as the main functional gene that could be specifically induced by EV71 vaccine.TASQNTLY,the amino acid sequence of CDR3,was identified as a specific binding site with the MHC-EV71 antigenic peptide.

关 键 词：T细胞受体 肠道病毒71型 疫苗 细胞免疫 体液免疫 

分 类 号：R392[医药卫生—免疫学]