肠癌RNA转染小鼠骨髓来源单核细胞的抗肿瘤效应

Effect on antitumor immunity of bone marrow-derived monocytes transfected with the RNA of murine carcinoma of colon

目的 :以提取的CT 2 6小鼠结肠癌细胞RNA作为抗原物质 ,体外转染骨髓来源的单核细胞 ,回输小鼠体内 ,观察其抗肿瘤效应 ;同时平行比较传统肿瘤冻融抗原体外冲击单核细胞的抗肿瘤效应。方法 :小鼠骨髓细胞体外以GM CSF诱导培养获取单核细胞 ,流式细胞仪检测纯度 ;Trizol法提取获得CT 2 6细胞总RNA ,应用TransMessenger体外转染单核细胞 ,对照组单核细胞以肿瘤冻融抗原冲击 ;LDH释放法检测小鼠体内CTL杀伤活性 ,观察各组小鼠成瘤情况及生存期。结果 :小鼠骨髓细胞经诱导培养后 ,获得大量高纯度的单核细胞 ,流式细胞仪检测CD11b+ >95 % ;提取的CT 2 6细胞总RNA体外经Trans Messenger介导转染单核细胞后 ,回输小鼠 ,可以诱导体内生成高水平的特异性细胞毒性T淋巴细胞 (CTL)活性 ,使小鼠获得抵抗后继CT2 6细胞攻击的免疫保护能力 ,并显著高于肿瘤冻融抗原冲击的单核细胞回输组。结论 :抗原提呈细胞经肿瘤总RNA转染后 ,可以诱导机体产生CTL 。

Objective:To investigate the effect on antitumor immunity of bone marrow-derived monocytes transfected with the RNA of CT-26(a cell line of murine carcinoma of colon), and meanwhile compared with the effect on antitumor immunity of thawed tumor antigens-pulsed monocytes.Methods:In vitro proliferation of the murine bone marrow-derived monocytes was stimulated by GM-CSF, the purity of which was detected by flow-cytometry. The total RNA of CT-26 was obtained by Trizol's process, with monocytes transfected by TransMessenger in vitro. The activity of cytotoxic T lymphocyte (CTL) in vivo was estimated by modified LDH release assay. Furthermore, the change of tumor's size in mice and animal's survival periods were also observed respectively in groups.Results:Plenty of monocytes were obtained from the culture of bone marrow, with over 95 percent positive rate of CD11b +. Vaccination with monocytes transfected with the total RNA could induce a high level of specific CTL activity in vivo and make mice resistant to the subsequent challenge of parental tumor cells. The in vivo effects induced by monocytes pulsed with the total RNA infection were more effective than those induced by monocytes pulsed with thawed tumor antigens.Conclusion:Antigen presenting cells transfected by the total RNA of tumor could present endogenetic tumor antigen,and could also activate CTL, thus inducing specific anti-tumor immunity effectively.