穿膜肽HIV-Tat_(49-57)和CTL表位融合多肽疫苗的初步免疫学效应研究  

Immunologic effect of a new peptide vaccine containing CTL epitope transferred by cell-penetrating peptide HIV-Tat_(49-57)

作　　者：王莉[1] 吴玉章[1] 石统东[1] 贾正才[1] 周伟[1] 邹丽云[1] 

机构地区：[1]第三军医大学基础医学部全军免疫学研究所,重庆400038

出　　处：《第三军医大学学报》2002年第10期1159-1161,共3页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划计划资助项目 ("973"项目 )(2 0 0 1CB510 0 0 1) ;国家自然科学基金资助项目 ( 3980 0 170 )

摘　　要：目的　探讨穿膜肽HIV Tat4 9- 57将CTL表位带入活细胞胞质并将其投入MHC Ⅰ类抗原提呈途径的可能性。方法　应用多肽固相合成技术 ,分别合成含HIVTat4 9- 57和HLA A2 1限制性CTL优势表位MART 12 7- 35的 18肽和该CTL表位 9肽。采用间接免疫荧光与激光共聚焦显微镜技术 ,分别对上述 18肽和 9肽的穿膜能力进行了动态观察。进一步用标准51 Gr释放试验分别检测了上述 18肽和 9肽在HLA A2 1阳性健康者外周血单个核细胞 (PBMC)中诱导的表位MART 12 7- 35特异性CTL活性。结果　 18肽能够穿过活哺乳动物细胞质膜进入胞质 ,并呈现出时间、剂量依赖关系 ;而 9肽则无此效应。与 9肽相比 ,18肽在体外诱导出了明显增强的特异性CTL活性 (P <0 0 5 )。结论　穿膜肽HIVTat4 9- 57可有效携带CTL表位穿过细胞膜进入胞质 ,并有效激发出针对该表位的特异性CTL应答 。Objective To discuss the possibilities of the intracellular delivery of CTL epitope by cell penetrating peptide HIV Tat 49-57 and of the ensuing MHC class Ⅰ pathway presentation. Methods A nonamer (P1) peptide (MART 1 27-35 ) of CTL epitote and an 18 mer peptide (P2) containing HIV Tat 49-57 and MART 1 27-35 were synthesized with aid of polypeptide solid phase synthesis technique. The intracellular transport capabilities of these peptides were tested with indirect immunofluorescence assay and confocal microscopy. In addition, the nonamer and 18 mer peptide were used to stimulate PBMC of the healthy HLA A2.1+ blood donors. Then, the activity of MART 1 27-35 specific CTL in PBMC of those donors was measured by using a standard 4 h 51 Gr release assay. Results The 18mer peptide was more effective than the nonamer in cellular uptake and efficiently crossed the plasma membranes of the live cells in a time and concentration-dependent fashion. Furthermore, the specific CTL activity induced by the 18mer was much stronger than that induced by the nonamer. Conclusion The results show that HIV Tat 49-57 can deliver the exogenous antigenic peptide into the cytoplasm of live cells and probably trigger the MHC class Ⅰ processing pathway. Thus, the described procedure should be applied to the design of peptide vaccine in order to induce CTL immunity against tumor and intracellular infection.

关 键 词：免疫学效应 穿膜肽 Tat49-57 CTL表位 MHC-I类抗原提呈途径 多肽疫苗 肿瘤免疫治疗 

分 类 号：R730.51[医药卫生—肿瘤] R392[医药卫生—临床医学]