反义B7-H1对人树突状细胞生物学特性的影响研究

目的研究反义B7-H1基因转染对人树突状细胞(dendritic cells,DCs)生物学特性的影响.方法以稳定表达反义B7-H1的重组腺病毒Ad.ASB7-H1转染DCs,转染后1、3、5、7及9 d以流式细胞术检测转染DCs表面B7-H1、CD1、CD14、D80、CD83及HLA-DR的表达,ELISA技术检测Ad.ASB7-H1转染DCs培养上清中IL-12的水平,3H-TdR掺入法检测Ad.ASB7-H1转染DCs诱导的T淋巴细胞增殖能力.结果反义B7-H1转染DCs后可明显抑制细胞表面B7-H1表达,其抑制效应在转染后24 h即开始出现.对DCs其他粘附分子表型表达无明显影响.Ad.ASB7-H1转染DCs刺激同种异体混合淋巴细胞反应能力增强;Ad.ASB7-H1转染DCs分泌IL-12能力提高.结论Ad.ASB7-H1转染可抑制DCsB7-H1的表达并使DCs生物学行为发生改变,反义B7-H1可能成为提高DCs疫苗生物学效能的一个潜在靶点.

Influence on biological characteristics of dendritic cells by transfecting B7-H1 antisense gene

Objective To investigate the effects of B7-H1 antisense gene transfection on the biological characteristics of dendritic cells (DCs) in human. Methods Human DCs were transfected with Ad.ASB7-H1 or Ad.LzcZ as positive control, and the untreated DCs served as normal control. Then the phenotypes of DCs, transfected with Ad.ASB7-H1 or Ad.LzcZ or normal, were detected of B7-H1, CD1, CD14, D80, CD83 and HLA-DR, with flow cytometry on post-transfection day 1, 3, 5, 7, 9. Enzyme-linked immunosorbent assay (ELISA) was used to detect IL-12 level of supernatant, 3H-TdR to detect the capacity of the DCs transfected of Ad.ASB7-H1 to activate T lymphocyte proliferation. Results The expression level of B7-H1 on transfected DCs was inhibited from 24 h after transfection, but the expression levels of CD1, CD14, D80, CD83 and HLA-DR were not significantly changed. Ad.ASB7-H1 transfection significantly increased the potency of secretion of IL-12 by transfected DCs and enhanced the ability of DCs to stimulate T cell proliferation. Conclusion DCs transfected by Ad.ASB7-H1 can inhibit B7-H1 expression and the biological characteristics of transfected DCs changed correspondingly. B7-H1 may be a promising target for improving the biological effects of DCs vaccine.