重组减毒细菌运送CD8^+T细胞表位的效应分析  

Analysis of in vitro delivering CD8^+ T epitopes by attenuated bacteria

作　　者：潘志明[1] 张晓明[1] 焦新安[1] Richard Lo-Man Claude Leclerc 刘秀梵[1] 

机构地区：[1]扬州大学农业部畜禽传染病学重点开放实验室,江苏扬州225009 [2]法国巴斯德研究所免疫调节生物学实验室,法国巴黎75015

出　　处：《细胞与分子免疫学杂志》2006年第6期730-733,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30425031);江苏省自然科学基金资助项目(BK2003043);扬州大学"新世纪人才工程"优秀青年骨干教师计划(2004年度)

摘　　要：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。AIM： To analyze the efficiency of delivery for CD8 ^＋ T cell epitopes by recombinant bacteria vectors. METHODS： The recombinant E. coil strain 13A or 25A and recombinant Salmonella typhimurium strain SL7207 expressing CD8 ^＋ T cell epitopes of Lymphocytic Choriomeningitis Virus （LCMV） and Ovalbumin （OVA）, which were fused with green fluorescent protein （GFP） marker at the C-terminal, were infected into LK^b cells, LL^d cells or bone marrow dendritic cells （BMDC）. The efficiency of presentation for CD8 ^＋ T cell epitopes by recombinant bacteria was analyzed by in vitro antigen presentation assay. RESULTS： After the infection of LK^b cells, LL^d cells or BMDC by recombinant bacteria, about 90% of cells were GFP positive. The results indicated that attenuated strain 13A, 25A and SL7207 had better invasive capacity to LK^b and LL^d cells, while BMDC had stronger capacity for uptaking recombinant bacteria. At 2 h post infection, CD8 ^＋ T cell epitopes presented on the surface of those LK^b, LL^d and BMDC cells infected by 13A （ptG2F） and SL7207（ptG2F） could be recognized by B3Z or nV1H7 T hybridoma cells, which were specific for OVA peptide p257-264 or LCMV peptide p118 - 132, respectively. But the efficiency of presentation for OVA or LCMV CD8 ^＋ T cell epitope was decreased or increased respectively at 48 h post infection. However, LK^b, LL^d and BMDC cells infected by 25A（ptG2F） did not effectively stimulate the specific B3Z or nV1 H7 T cells. Furthermore, the presentation efficiency of BMDC was higher than those of LK^b and LL^d cells under the same condition. CONCLUSION： CD8 ^＋ T cell epitopes can be delivered and presented to T lymphocytes by APC by attenuated bacteria in vitro.

关 键 词：CD8^+T细胞表位 减毒大肠杆菌 减毒鼠伤寒沙门氏菌 体外抗原提呈试验 

分 类 号：R392.12[医药卫生—免疫学]