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目的:探讨HER2/neu蛋白细胞胞外段的基因重组腺相关病毒(rAAV-HER2/neu)和粒-巨噬细胞集落刺激因子重组腺相关病毒(rAAV-GM-CSF)共转染树突状细胞(DC)诱导的细胞免疫应答。方法:成熟的DC通过淋巴细胞分离液分离培养获得,流式细胞技术分析HER-2/neu以及细胞表面标志CD1a、CD83、CD80、CD86和HLA-DR的表达,MTT检测混合淋巴反应,3H-TdR释放法检测CTL杀伤活性,RT-PCR检测FasL表达。结果:共转染组(CT)、单独转染组(ST)和未转染组(UT)3组DC的细胞表面标志表达无统计学意义(P>0.05),共转染组中HER2/neu和FasL的表达比单独转染组高,共转染组中可以检测到更强的混合淋巴细胞反应和CTL杀伤活性。结论:HER2/neu蛋白细胞外段的基因通过DC的抗原提呈作用可以诱导出免疫应答,其免疫应答作用与FasL表达相关,而GM-CSF可以增强其免疫应答作用。
Objective: To investigate the expression of dendritic cells (rAAV-HER2 / neu) and granulocyte-macrophage colony stimulating factor recombinant adeno-associated virus (rAAV-GM-CSF) in the extracellular domain of HER2 / Cell (DC) -induced cellular immune response. Methods: The mature DCs were isolated and cultured by lymphocyte separation. The expression of HER-2 / neu and cell surface markers CD1a, CD83, CD80, CD86 and HLA-DR were analyzed by flow cytometry. TdR release assay CTL killing activity, RT-PCR detection FasL expression. Results: There was no significant difference in cell surface markers between DCs transfected (CT), ST alone and untransfected (UT) DCs (P> 0.05) The expression of neu and FasL was higher than that of the transfection alone group, and stronger mixed lymphocyte reaction and CTL killing activity could be detected in the co-transfection group. Conclusion: The extracellular domain of HER2 / neu protein can induce immune response by DC antigen presentation. The immune response is related to FasL expression, while GM-CSF can enhance its immune response.