中华皮肤科杂志 ›› 2004, Vol. 37 ›› Issue (4): 194-496.

• 论著 • 上一篇    下一篇

负载人乳头瘤病毒16型E7多肽树突细胞诱导抗原特异性细胞免疫反应

黄伟峰, 刁庆春, 郝飞, 徐云升, 钟白玉, 叶庆佾   

  1. 第三军医大学附属西南医院皮肤科 重庆 400038
  • 收稿日期:2003-04-27 出版日期:2004-04-15 发布日期:2004-04-15
  • 通讯作者: 郝飞,E-mail:haofei@mail.tmmu.com.cn E-mail:haofei@mail.tmmu.com.cn
  • 基金资助:
    国家自然科学基金资助课题(30070698)

Antigen-specific Cellular Immune Response Induced by Dendritic Cells Activated by Human Papilloma Virus 16 E7 Peptide

HUANG Wei-feng, DIAO Qing-chun, HAO Fei, XU Yun-sheng, ZHONG Bai-yu, YE Qing-yi   

  1. Department of Dermatology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
  • Received:2003-04-27 Online:2004-04-15 Published:2004-04-15

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摘要： 目的 观察负载人乳头瘤病毒16型(HPV16)E749-57多肽树突细胞(DC)体外诱导针对HPV16+肿瘤细胞的特异性细胞免疫反应.方法 用粒细胞-巨噬细胞集落刺激因子(GM-CSF)及白介素4(IL-4)从健康成人外周血单一核细胞诱导DC并负载HPV16E749-57多肽,流式细胞仪检测DC表面抗原;混合淋巴细胞反应观察DC对同种淋巴细胞增殖的激发效应;乳酸脱氢酶(LDH)释放法评价DC诱导的细胞毒性T淋巴细胞(CTL)对Caski肿瘤细胞体外杀伤效应.结果 负载HPV16E749-57多肽的DC表面表达CD1a(58.4±6.7)%、CD80(70.6±3.4)%及HLA-DR(74.8±4.2)%分子;具有刺激同种T淋巴细胞增殖的能力;其诱导的抗原特异性CTL对Caski肿瘤细胞产生特异性杀伤,而对SiHa、AN3CA肿瘤细胞无明显杀伤作用.结论 负载HPV16E749-57多肽的DC体外可诱导高效、特异性的CTL效应.

关键词: 乳头状瘤病毒,人, 树突细胞, T淋巴细胞,细胞毒性, E7抗原

Abstract: Objective To investigate specific cellular immune response induced by dendritic cells(DCs) activated by human papilloma virus (HPV) 16 E7 peptide. Methods DCs separated from peripheral blood mononuclear cells were induced with granulocyte/macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4), the DCs were then activated by HPV 16 E7 peptide. After that the surface antigens on the DCs were detected by flow cytometer. Homogenic mixed lymphocyte reaction (HMLR) with DCs was performed. LDH release assay was also used to detect the activity of cytotoxic T lymphocytes(CTL)to Caski tumor cells, which was induced by DCs. Results The surface antigens on the activated DCs were highly expressed, such as CD1a(58.4±6.7)%, CD80(70.6±3.4)% and HLA-DR(74.8±4.2)%. HMLR showed that the activated DCs stimulated T cell proliferation significantly. Antigen-specific CTL induced by the activated DCs specifically killed Caski tumer cells, but had no effect on SiHa and AN3CA cells. Conclusions DCs loaded with HPV 16 E7 peptide can induce highly effective and specific cellular immune response.

Key words: Papillomavirus, human, Dendritic cells, T lymphocytes, cytotoxic, E7 antigen