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目的探讨新城疫病毒(newcastlediseasevirus,NDV)对Hep-2肿瘤细胞的杀伤效应及其免疫刺激作用。方法采用考马斯亮兰染色法观察NDV感染对Hep-2肿瘤细胞细胞骨架的影响。运用甲基噻唑基四唑(methylthiazolyltetrazolium,MTT)染色法检测NDV感染致Hep-2肿瘤细胞死亡的杀伤率。通过免疫荧光结合流式细胞仪(flowcytometer,FCM)检测Hep-2肿瘤细胞表面主要组织相容性复合体Ⅰ类分子(majorhistocompabilitycomplexclassImolecules,MHC-I)的表达情况。另外,分离人外周血淋巴细胞(peripheralbloodmononuclearcells,PBMC)作为效应细胞,以NDV感染的Hep-2肿瘤细胞作为靶细胞,同时设空白细胞对照,通过特异性细胞毒T淋巴细胞实验,探讨NDV所介导的抑瘤作用。结果NDV可致Hep-2肿瘤细胞的细胞骨架发生改变,对体外培养Hep-2肿瘤细胞的杀伤率达77.7%,可上调其表面MHC-I类分子的表达,并且能够刺激增强人外周血淋巴细胞中肿瘤特异性细胞毒性T淋巴细胞(cytotoxicTlymphocyte,CTL)活性,杀伤率为23.54%。结论NDV可有效杀伤Hep-2肿瘤细胞,并刺激产生特异性抗肿瘤免疫反应。
Objective To investigate the killing effect of Newcastle disease virus (NDV) on Hep-2 tumor cells and its immunostimulatory effects. Methods Coomassie brilliant blue staining was used to observe the effect of NDV infection on the cytoskeleton of Hep-2 tumor cells. The death rate of Hep-2 tumor cells induced by NDV was detected by methylthiazolyltetrazolium (MTT) staining. The expression of major histocompatibility complex class I molecules (MHC-I) on Hep-2 tumor cells was detected by immunofluorescence combined with flow cytometry (FCM). In addition, human peripheral blood mononuclear cells (PBMCs) were isolated as effector cells and NDV-infected Hep-2 tumor cells as target cells. At the same time, blank control cells were established. Through specific cytotoxic T lymphocyte assay, Anti-tumor effect guide. Results NDV induced a change in the cytoskeleton of Hep-2 tumor cells, killing 77.8% of Hep-2 tumor cells in vitro and up-regulating the expression of MHC class I molecules on the surface of Hep-2 tumor cells, The cytotoxic T lymphocyte (CTL) activity in the cells was 23.54%. Conclusion NDV can effectively kill Hep-2 tumor cells and stimulate specific anti-tumor immune response.