论文部分内容阅读
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 analyze the clonalization of CD4 + TCR Vβ gene family in BALB / c mice induced by EV71 vaccine and to explore the immune mechanism of EV71 vaccine. Methods The clonalization of CD4 + TCRVβ gene family in BALB / c mice induced by EV71 vaccine was analyzed by TCR cloning assay kit and gene scanning technique. The EV71 vaccine-induced clonalization of the CD4 + TCRVβ gene family was analyzed by ELISPOT method, Luminex technique and in vitro micro- Cellular immunity and humoral immune response to explore the EV71 vaccine immune mechanism. Results Clone of Vβ1-20 in mouse CD4 + TCR showed that there was a monoclonal or oligoclonal change in the Vβ17 gene family in the immunized group and that in the control group, polyclonal Vβ17 was not observed. The secretion of IFN-γ and IL-6 in spleen MNC (excluding CD8 +) in the immunized group was significantly higher than that in the control group (all P <0.01), and the neutralizing antibody of serum EV71 was all positive. Further sequencing revealed that the amino acid sequence of Vβ17 in the immunized group was TASQNTLY. Conclusion The Vβ gene family of EV71 vaccine-induced CD4 + TCR-specific changes in BALB / c mice was successfully selected as Vβ17. The amino acid sequence of the CDR3 region specifically binding to the MHC-EV71 antigen peptide was TASQNTLY.