中华皮肤科杂志

• 论著 • 上一篇    下一篇

HIV Tat49-57增强人乳头瘤病毒16E7细胞毒性T细胞表位穿膜效应研究

尹锐, 郝飞, 郝进, 钟白玉, 曹红卫   

  1. 第三军医大学西南医院皮肤科 重庆 400038
  • 收稿日期:2005-05-19 出版日期:2006-04-15 发布日期:2006-04-15
  • 通讯作者: 郝飞,email:haofei@mail.tmmu.com.cn E-mail:haofei@mail.tmmu.com.cn
  • 基金资助:
    国家自然科学基金(30070698，30500609)

Cell-penetrating effects of HPV16E749-57 epitope modified by peptide HIV-Tat49-57

YIN Rui, HAO Fei, HAO Jin, ZHONG Bai-yu, CAO Hong-wei   

  1. Department of Dermatology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
  • Received:2005-05-19 Online:2006-04-15 Published:2006-04-15

PDF (PC)

75

赞

摘要： 目的 探讨有穿膜序列HIV Tat49-57的人乳头瘤病毒(HPV)16E7限制性细胞毒性T细胞(CTL)表位融合肽的跨膜能力及其影响因素。方法 在HPV16E7 HLA-A2+/H-2kb+限制性CTL表位E749-57的N末端连接穿膜肽HIV Tat49-57序列,应用固相技术合成此融合肽,用间接免疫荧光与激光共聚焦显微技术,研究其穿膜能力,用乳酸脱氢酶(LDH)释放法检测其体外诱导特异性CTL杀伤效应。结果 穿膜序列HIV Tat49-57可以有效促进HPV16E749-57表位肽进入BHK细胞胞浆,并呈时间、剂量依赖关系(P<0.05);与原表位肽相比,该融合肽有显著地激发特异性CTL活性的能力。结论 在原CTL表位基础上引入穿膜序列,可以使其更快速、更有效地进入活细胞胞浆,为HPV16肽疫苗的设计提供一种新的思路。

关键词: 基因产物, tat, 表位, T淋巴细胞, 蛋白质转运, 疫苗, 亚单位

Abstract: Objective To investigate the effects of the intracellular delivery of HPV16E749-57 epitope modified by cell-penetrating peptide HIV-Tat49-57 and its influential factors.Methods The unique HLA-A2+/H-2kb+ limited CTL epitope of HPV16E7 fused with a cell-penetrating sequence (HIV-Tat49-57) was designed and a 18-mer peptide was synthesized with aid of polypeptide solid phase synthesis technique. The intracellular transport capabilities of these peptides were tested with indirect immunofluorescence assay and laser confocal microscopy.In addition,the CTL epitope and 18-met peptide were used to stimulate PBMC of healthy C57BL/6 mice,and the E749-57 specific CTL responses were measured by LDH cytotoxicity detection kit in PBMC of those mice.Results The HIV-Tat49-57 peptide could efficiently assist HPV 16E749-57 peptide in penetrating into BHK cells in a time and dose-dependent manner (P<0.05).Furthermore,the specific CTL activity induced by the 18-mer peptide was much stronger than that induced by the single epitope.Conelusion The results show that HIV-Tat49-57 can efficiently deliver the exogenous antigenic peptide into the cytoplasm of live cells,and induce specific CTL response.This is a new way to design peptide-based vaccine of HPV.

Key words: Gene products,tat, Epitopes,T-lymphocyte, Protein transport, Vaccines,subunit