腺病毒介导的体外免疫方法研究

A research of adenovirus-mediated in vitro immunization

目的:探讨腺病毒介导的体外免疫方法。方法:本实验中主要通过建立小鼠骨髓源树突状细胞体外培养方法,在此基础上用携带绿色荧光蛋白(GFP)报告基因的腺病毒转导树突状细胞(DC),通过荧光倒置显微镜观察荧光表达,并经流式细胞仪检测GFP平均荧光强度表达以检测腺病毒转导效率。穿刺法制备小鼠脾单细胞悬液,腺病毒转导树突状细胞与脾细胞以1:10比例进行混合细胞培养后,经多肽(HYLSTQSAL)刺激通过流式细胞仪检测T淋巴细胞IFN-γ的分泌。结果:经多肽刺激后流式细胞仪可检测到T淋巴细胞IFN-γ的分泌。结论:成功建立腺病毒介导树突状细胞(DC)抗原呈递的体外免疫方法。

Objective：To evaluate the adenovirus-mediated immunization in vitro.Methods：In this experiment,the cultivation of mouse bone marrow-derived dendritic cells were established in vitro,and on this basis,dendritic cells were transduced by adenoviruses which carry the green fluorescent protein（GFP） reporter gene.By inverted fluorescence microscope the expression of fluorescence were observed,and by flow cytometry the mean fluorescence intensity of GFP were detected in order to detect the transduction efficiency of adenovirus.The mouse spleen cell suspension was prepared through puncture.Dendritic cells were transduced by adenoviruses and were mixedly cultured with spleen cells in proportion of 1：10.By flow cytometry,the IFN-γ secretion was detected after T lymphocytes were stimulated with polypeptide（HYLSTQSAL）.Results：The IFN-γ secretion can be detected by flow cytometry after T lymphocytes were stimulated with polypeptide.Conclusion：The immune method in vitro that adenovirus mediates antigen-presenting of dendritic cells has been successfully established.