中华皮肤科杂志

• 论著 • 上一篇    下一篇

穿膜肽对HPV16E7细胞毒性T细胞表位MHC-Ⅰ类抗原提呈影响的初步研究

尹锐, 郝飞, 李芹阶, 夏汝山, 钟白玉   

  1. 第三军医大学西南医院皮肤科, 重庆400038
  • 收稿日期:2006-12-19 出版日期:2007-11-15 发布日期:2007-11-15
  • 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(30500609)

Effects of cen-penetrating peptide on the presentation of MHC classⅠ-restricted cytotoxic T lympho-cyte epitope of human papillomavirus 16E7

YIN Rui, HAO Fei, LI Qin-jie, XIA Ru-shan, ZHONG Bai-yu   

  1. Department of Dermatology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
  • Received:2006-12-19 Online:2007-11-15 Published:2007-11-15

摘要: 目的 研究穿膜肽(Tat49-57)对HPV16E7 MHC-Ⅰ类限制性细胞毒性T细胞(CTL)表位(E749-57)在抗原提呈细胞内经MHC-Ⅰ类途径被提呈的动力学影响。方法 应用多肽固相合成技术,分别合成含穿膜肽与HPV(人乳头瘤病毒)16E7惟一HLA-A2/H2-Kb+限制性CTL表位E749-57的融合多肽,同时合成该表位N端自然延伸的抗原肽和无关对照肽。采用流式细胞仪(FACS)分析技术,检测抗原提呈细胞(APE)在不同时相点对上述各抗原肽进行MHC-Ⅰ类提呈的情况。结果 在与APC孵育早期,Tat-E749-57在短时间内即被APC快速提呈并进入MHC-Ⅰ类抗原提呈途径,提呈的效率略低于E749-57(P>0.05);在孵育的后期,与Tat-E749-57组孵育的APC细胞表面,E749-57/Kb复合物存在时间较其他组明显延长(P<0.05)。结论 在外源性抗原肽中引入穿膜肽可明显促进其MHC-Ⅰ类限制性CTL表位的提呈效率,从而有效增强外源性抗原肽的免疫原性。

关键词: 乳头状瘤病毒, 人, 抗原呈递, 穿膜肽, CTL表位

Abstract: Objective To investigate the kinetic effects of cell-penetrating peptide (CPP) Tat49-57 on the presentation of human papillomavirus (HPV) 16E749-57,a MHC class I-restricted epitope of cytotoxic T lymphocytes (CTL).Methods A fusion protein containing CPP Tat49-57 and E749-57,the unique HLA-A2/H2-Kb+ restricted CTL epitope of HPV16E7,was synthesized by standard solid-phase Fmoc chemistry.A N-terminal extended epitope and an irrelated peptide were synthesized respectively as controls.A macrophage cell line Ana-1 was cultured and incubated with the antigen peptides for various durations.Fluorescence activated cell sorter (FACS) was employed with FITC-labeled monoclone antibody to E749-57/Kb to detect the expression of E749-57 on the surface of Ana-1 cells.Results At the early stage (within 80 min) of incubation,Tat-E749-57 could be uptaken by APCs and presented by MHC classⅠmolecules much more rapidly than other antigen peptides,but its presentation effiency was relatively lower as compared with E749-57 (P>0.05).At the late stage (8-48 h) of incubation,the presence time of E749-57/Kb was significantly prolonged on the surface of Tat-E749-57-incubated cells than that on the surface of other peptides-incubated cells (all P<0.05).Conclusion CPP can effectively enhance the immunogenicity of exogenous antigen peptides by promoting MHC classⅠantigen presentation of CTL epitopes.

Key words: Papillomavirus,human, Antigen presentation, Cell-penetrating peptides, CTL epitope