MUC1体外转染脐带血DC疫苗抗乳腺肿瘤的免疫效应研究

目的:研究人粘蛋白核心肽-连续重复序列(MUC1-VNTR)基因转染的脐血DC疫苗抗乳腺肿瘤的免疫效应。方法:提取人脐带血淋巴细胞,培养脐血树突状细胞(DCs),采用Lipofectamine2000阳离子脂质体法将MUC1-VNTR转染入脐血DC,用Western印迹法检测MUC1-VNTR基因的表达;与自体T细胞共培养,用MTT法检测转染MUC1基因的脐血DC所致敏的细胞毒性淋巴细胞对乳腺癌MCF-7的杀伤活性;ELISA法检测其刺激T细胞分泌IFN-γ的能力。结果:Western印迹法检测到1条MUC1-VNTR基因表达的特殊条带,转染MUC1-VNTR基因的脐血DC对MUC1表达阳性的乳腺癌细胞系MCF-7的杀伤活性显著高于单纯脐血DC组(P<0.05),而且其刺激同种T细胞分泌高水平的IFN-γ,显著高于未转染的脐血DC(P<0.05)。结论:MUC1-VNTR基因转染的人脐血树突状细胞可诱导特异性CTL,产生有效的针对MUC1阳性乳腺癌细胞MCF-7的杀伤效应,并诱导特异性抗乳腺癌免疫应答。

Study on immunological effect of umbilical cord blood dendritic cell vaccine transfected by MUC1 in vitro on breast tumor

Objective:To research the immunological effect of umbilical cord blood dendritic cell(DC) vaccine transfected by MUC1-VNTR gene in vitro on breast tumor.Methods:Human umbilical cord blood lymphocytes were Abstracted,then umbilical cord blood dendritic cells were cultured,Lipofectamine 2000 cationic liposome method was used to transfect MUC1-VNTR into umbilical cord blood dendritic cells,Western blot was used to detect the expression of MUC1-VNTR gene;umbilical cord blood dendritic cells were cocultured with autologous T lymphocytes,MTT method was used to detect the cytotoxicity of cytotoxic lymphocytes sensitized by umbilical cord blood dendritic cells after transfection of MUC1 gene on breast cancer MCF-7 cell line;ELISA was used to detect its stimulating ability on IFN-γ secretion of T lymphocytes.Results:One special band with MUC1-VNTR gene expression was detected by Western blot,the cytotoxicity of cytotoxic lymphocytes sensitized by umbilical cord blood dendritic cells after transfection of MUC1-VNTR gene on breast cancer MCF-7 cell line with positive expression of MUC1 was statistically significantly stronger than that sensitized by simple umbilical cord blood dendritic cells(P<0.05),and its stimulating ability on IFN-γ secretion of T lymphocytes was statistically significantly higher than sensitized by simple umbilical cord blood dendritic cells(P<0.05).Conclusion:Human umbilical cord blood dendritic cells transfected by MUC1-VNTR gene can induce specific cytotoxic lymphocytes,produce effective cytotoxic effect targeting breast cancer MCF-7 cell line with positive expression of MUC1,and induce specific immunologic response of anti-breast cancer.