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目的:探讨肿瘤细胞RNA转染人脐血树突状细胞分泌的exosomes对细胞毒T细胞特异性抗肿瘤作用的影响。方法:以人脐血单个核细胞为来源,应用干细胞因子、粒细胞集落刺激因子、白细胞介素4诱导和BGC823RNA转染形成成熟树突状细胞;收集树突状细胞培养上清,超速离心法分离exosomes,并用透射电镜进行鉴定;利用MTT比色分析法观察exosomes诱导T细胞增殖和细胞毒T细胞的杀瘤活性。结果:exosomes激活的细胞毒T细胞对BGC-823细胞的杀伤率分别为(38.13±9.67)%和(50.72±11.23)%,与未经exosomes激活的细胞毒T细胞相比差异有统计学意义(P<0.05)。结论:经肿瘤RNA转染后的人脐血诱导的树突状细胞分泌的exosomes在体外能激活细胞毒T细胞对肿瘤细胞的杀伤活性,exosomes在非细胞性疫苗生物治疗方面有广阔前景。
Objective: To investigate the effect of exosomes secreted by tumor cell RNA transfected human dendritic cells on cytotoxic T lymphocyte-specific antitumor effect. Methods: Human umbilical cord blood mononuclear cells were used as source of stem cell factor, granulocyte colony-stimulating factor, interleukin-4 and transfected with BGC823RNA to form mature dendritic cells. The dendritic cell culture supernatants were collected and purified by ultracentrifugation The exosomes were isolated and identified by transmission electron microscopy. The cytotoxic activity of exosomes on T cell proliferation and cytotoxic T lymphocytes was observed by MTT colorimetric assay. RESULTS: The killing rates of exosomes-activated cytotoxic T cells to BGC-823 cells were (38.13 ± 9.67)% and (50.72 ± 11.23)%, respectively, which were significantly different from those of cytotoxic T cells without exosomes activation (P <0.05). CONCLUSION: Exosomes secreted by dendritic cells induced by human umbilical cord blood after tumor RNA transfection can activate the cytotoxic activity of cytotoxic T cells against tumor cells in vitro. The exosomes have broad prospects in the biological treatment of non-cellular vaccines.