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目的:分析重组减毒菌体外运送CD8+T细胞表位的效应。方法:以表达卵清蛋白(OVA)和淋巴细胞脉络丛脑膜炎病毒(LCMV)CD8+T细胞表位的重组菌13A(ptG2F)、25A(ptG2F)和SL7207(ptG2F)感染抗原提呈细胞(APC)LKb、LLd和骨髓源树突状细胞(BMDC),应用体外抗原提呈试验检测APC对重组菌运送的CD8+T细胞表位的提呈效应。结果:感染试验证实,减毒菌13A、25A和SL7207对LKb细胞或LLd细胞均具有良好的侵袭能力,BMDC对重组菌具有很好的摄取功能。抗原提呈试验结果显示,在感染的早期(2h),LKb、LLd细胞和BMDC均可提呈重组菌13A(ptG2F)或SL7207(ptG2F)运送的T细胞表位;在感染的晚期(48h),LKb细胞对OVA257-264CD8+T细胞表位的提呈效应降低,LLd细胞对LCMV118-132CD8+T细胞表位的提呈效应增强。3种APC均不能提呈25A(ptG2F)运送的T细胞表位。另外,在同样的作用条件下,BMDC对减毒菌运送的抗原表位的提呈效应要强于LKb和LLd细胞。结论:重组菌能运送CD8+T细胞表位,为基于减毒细菌的新型基因工程疫苗的分子设计提供了有益借鉴。
Objective: To analyze the effect of recombinant attenuated bacteria carrying CD8 + T cell epitopes in vitro. METHODS: Antigen-presenting cells (n = 8) were infected with recombinant strains 13A (ptG2F), 25A (ptG2F) and SL7207 (ptG2F) expressing OVA and LCMV CD8 + APC) LKb, LLd and bone marrow-derived dendritic cells (BMDC). The in vitro antigen-presenting test was used to detect the APC-presenting effect on CD8 + T cell epitopes delivered by recombinant bacteria. Results: Infection experiments confirmed that attenuated bacteria 13A, 25A and SL7207 had good invasion ability on LKb cells or LLd cells, and BMDC had a good uptake function for the recombinant bacteria. The results of antigen presentation showed that both LKb, LLd cells and BMDCs could express T cell epitopes delivered by recombinant strain 13A (ptG2F) or SL7207 (ptG2F) in the early stage of infection (2 h) , LKb cells decreased the presentation of OVA257-264CD8 + T cell epitopes, and LLd cells presented an enhanced effect on LCMV118-132CD8 + T cell epitopes. None of the three APCs presented the 25A (ptG2F) delivered T cell epitope. In addition, under the same conditions, the presentation of BMDC to antigenic epitopes delivered by attenuated bacteria was stronger than that of LKb and LLd cells. CONCLUSION: Recombinant bacteria can deliver CD8 + T cell epitopes, which provides a useful reference for the molecular design of novel genetic engineering vaccines based on attenuated bacteria.