永生化EB病毒转化B细胞对肝癌细胞抗原的提呈作用

Antigen presenting effect of immortalized EBV-transformed B-lymphoblastoid c ells on tumor antigen

目的 建立EB病毒 (Epstein Barrvirus,EBV)转化的永生化B细胞 (命名为LEBV)并研究其对肝癌细胞抗原的提呈作用。方法 从人外周血中分离出外周血单个核细胞 (Peripheralbloodmononuclearcell,PBMC) ,用等体积的EBV上清混合培养 4周以上建立永生化EBV转化B细胞 (LEBV) ,用流式细胞计数仪 (FACS)检测其细胞表面功能相关分子CD86、CD4 0、HLA DR和HLA ABC的表达情况。LEBV与自体淋巴细胞和肝癌细胞抗原共培养 3d,结束培养前 1 2h加入 37kBq/孔3 H 标记的胸腺嘧啶 (3 H TdR) ,收获细胞 ,用液体闪烁计数仪测定cpm。结果 培养 3周以上即培养出EBV转化的永生化B细胞 ,可连续培养 1年以上。经FACS检测 ,细胞表面分子CD86、CD4 0、HLA DR和HLA ABC的表达率分别为 74 %、91 %、83%和 86 .7%。该细胞接触肝癌细胞抗原后刺激自体淋巴细胞增殖的能力增强。

Objective To set up immortalized EBV transformed B lymp hoblastoid cells and investigate their ability of presenting antigen of hepatoca rcinoma cells.Methods Isolated peripheral blood mononuclear cel ls (PBMC) from human peripheral blood cultured with same volume super natant of EBV were analysed their function related molecules CD86,CD40,HLA D R and HLA ABC by FACS. Cultured LEBV with autologus lymphocyte and antig en of Hepatocarcinoma for three days and added 37 kBq/well 3H TdR 12 hours before culture completion. Cpm was detected.Results Im mortalized EBV transformed B lymphoblastoid cells were obtained after three we eks.The y could survive more than one year. The expression of CD86,CD40,HLA DR and HL A ABC of them were 74%,91%,83% and 86.7%.The ability of stimulation autologou s ly mphocytes proliferation of cells were better than original cells. Conclu sion Immortalized EBV transformed B lymphoblastoid cells were able to present antigen of Hepatocarcinoma cells.