Tα1对食管癌细胞RNA转染的脐血树突状细胞功能的影响

The effect of Tα1 on human cord blood dendritic cells transfected with RNA of esophageal carcinoma cells

目的探讨食管癌T.Tn细胞RNA转染脐血树突状细胞(DCs)对T细胞增殖和细胞毒性T淋巴细胞(CTL)特异性抗肿瘤作用的影响,以及胸腺肽α1(Tα1)对脐血DCs疫苗的免疫佐剂作用。方法分离脐血单个核细胞,经重组人粒细胞巨噬细胞集落刺激因子(rhGM-CSF)、重组人干细胞因子(rhSCF)、重组人IL-4(rhIL-4)诱导和T.Tn细胞RNA转染形成成熟DCs,加入Tα1制备肿瘤疫苗。检测RNA转染后DCs表型变化;MTT法检测各组脐血DCs诱导T细胞增殖和CTL细胞毒活性。结果与转染前比较,T.Tn细胞RNA转染后脐血DCs表面MHC-Ⅰ、MHC-Ⅱ和CD54、CD80和CD86分子水平明显升高(P均<0.05);可显著促进T细胞增殖,有效诱导CTL的特异性杀瘤活性(P<0.05)。而Tα1能显著促进RNA致敏脐血DCs的表面分子表达和T细胞功能(P<0.05)。结论Tα1联合食管癌细胞RNA转染脐血DCs可制备高效、特异的肿瘤疫苗,有望成为食管癌生物治疗的新途径。

Objective To investigate the proliferation of T lymphocyte and antitumor activity of CTL induced by human cord blood dendritic cells （DCs） transfected with RNA of esophageal carcinoma, and immuno-adjuvant effect of Tα1 on DCs. Methods The RNA of T. Tn cells were prepared by Trizol reagent. The cord blood monocytes were cultured to produce DCs with rhSCF, rhGM-CSF and rhIL-4. The DCs were collected and transfected with tumor cell total RNA. Tα1 was added in culture system to enhance the DCs vaccine. MHC- Ⅰ , MHC-Ⅱ and co-stimulatory molecules, CD54, CD80 and CD86 on the surface of DCs were analyzed by FCM. The Mixed lymphocyte reation （MLR） and cytotoxicity of CTLs to T. Tn cells were assayed by MTT colorimetry. Results DCs transfected with RNA of esophageal carcinoma ceils expressed more MHC- Ⅰ , MHC- Ⅱ , CD54, CD80 and CD86 than that without transfection （P 〈 0.05）. Mature transfected DCs could promote the proliferation of T lymphocyte and induce specific activity of CTL. Tα1 can enhance the expression of the molecules on surface and the activity of T lymphocyte significantly. Conclusion The vaccine can be prepared by cord blood DCs transfected with Tα1 and RNA of esophageal carcinoma cells, and it may provide effective and specific way for immuno-therapy of esophageal carcinoma.