6B11抗独特型微抗体负载树突状细胞对卵巢癌细胞系细胞杀伤作用的体外研究

In vitro study on cytotoxicity of T cells induced by dendritic cells loaded with 6B11 anti-idiotype minibody to ovarian cancer cell lines

目的 用 6B1 1抗独特型微抗体体外负载树突状细胞 (DC) ,以期诱导出抗原特异性的抗肿瘤免疫反应。方法 分离培养HLA -A2 +健康人外周血树突状细胞 ,培养过程中用 6B1 1微抗体负载 ,无关抗原F (ab)’2负载及未负载组为对照。光镜、电镜下观察树突的形态 ;流式细胞仪检测DC表面分子表达 ;3 H -TdR掺入法测DC刺激自体T细胞增殖 ;51 Cr 6h释放试验测激活的T细胞对卵巢癌细胞系的杀伤。结果 培养的树突细胞有典型形态 ,CD80、CD86、HLA -DR等表面分子呈高表达 (分别为 6 5 %、 86 2 %、 93 7% ) ;6B1 1微抗体负载的DC不仅能刺激自体T细胞的增殖 ,而且其诱导的CTL细胞对HLA -A2 +、OC1 6 6 - 9+卵巢癌细胞系HOC1A有特异性的杀伤 ,结论 6B1 1抗独特型微抗体模拟卵巢癌抗原OC1 6 6 - 9负载树突状细胞在体外可以诱导出抗原特异性的细胞毒T细胞 ,为进一步研究 6B1

Objective To observe whether dendritic cells (DCs)loaded with 6B11 anti-idiotype minibody can induce antigen specific anti-tumor reponses in vitro.Methods Peripheral blood monocytes were seperated from two HLA-A2 positive heathy volunteers and DCs were cultured from it.The immuture DCs were pulsed with 6B11 anti-idiotype minibody,and DCs unpulsed or pulsed with non-related mouse F(ab)'2 fragments served as controls.Morphology of the DCs was observed under microscope and electronic microscope.Cell surface molecules were examined by FACS. 3H-thymidine incorparation were employed to observed whether the DCs could stimulate proliferation of autologous T cell. 51 Cr 6-hour release test was used to estimated cytotoxicity of T cells activated by DCs to ovarian cancer cell lines in antigen specific ways.Results DCs raised in our way showed typical morphology as described by others.CD80、CD86、HLA-DR were over expressed,65%,86 2% and 93 7% respectively.DCs pulsed with 6B11 minibody could not only stimulate autologous naive T cells proliferation,but induce higher specific cytotoxicy to HOC1A cells,which were both HLA-A2 and OC166-9 positive expressed.Conclusions DCs loaded with 6B11 anti-idiotype minibody,which mimic ovarian carcinoma antigen OC166-9,can induce antigen specific T cell in vitro.This give us inspiration to further investigation on immunotherapy of 6B11 anti-idiotype minibody in ovarian carcinoma.