B7-1和IL-12基因转染对肝癌细胞免疫原性的影响

IMMUNE RESPONSES INDUCED BY HepG2 CELLS EXPRESSING B7-1 OR IL-12 MOLECULES

目的 观察B7 1和IL 12基因表达对人肝癌细胞免疫原性的影响。方法 分别将B7 1和IL 12基因以逆转录病毒介导转染HepG2细胞。阳性克隆细胞与健康人外周血淋巴细胞 (PBL)混合培养后 ,用流式细胞仪检测PBL表面Ⅰ类人白细胞抗原 (HLA Ⅰ )分子表达 ,以MTT法检测PBL的特异性杀伤活性及杀伤K5 6 2细胞的活性。结果 混合培养后HepG2 /B7 1和HepG2 /IL 12细胞组PBL表面的HLA Ⅰ分子表达分别较HepG2 /neo细胞组增加16 .95 %和 14 .71% (P <0 .0 5 ) ;与HepG2 /neo组比较 ,HepG2 /B7 1组PBL特异性杀伤活性增高 12 .5 % (P <0 .0 5 ) ,HepG2 /IL 12组PBL的特异性杀伤活性无明显增高 (P >0 .0 5 )。HepG2 /B7 1、HepG2 /IL 12细胞活化的PBL对K5 6 2细胞的杀伤活性较HepG2 /neo组分别增高 19.38%和 14 .78% (P <0 .0 5 )。 结论 B7 1和IL 12分子均能增强免疫细胞对肝癌细胞的识别和杀伤 。

Objective To investigate the effect of B7 1 and IL 12 gene expression on the immunogenicity of hepatocellular carcinoma (HCC) HepG2 cells. [WT5”HZ]Methods [WT5”BZ]Plasmids encoding B7 1 and IL 12 molecules were retrovirally introduced into human HCC cells,empty vector as control. PBLs were cocultured with HepG2/B7 1,HepG2/IL 12 and HepG2/neo cells. Three days later,PBLs were submitted to specific cytotoxicity test and nonspecific cytotoxicity test against K562 cells by MTT assay.[WT5”HZ]Results [WT8.75BZ] HLA Ⅰ molecules on PBLs were detected by FACS.HLA Ⅰ molecules expressing on PBL cocultured with HepG2/B7 1,HepG2/IL 12 cells were enhanced by 16.95% and 14.71% than those of HepG2/neo group, respectively( P <0.05). Specific cytotoxicity against HepG2/B7 1 cells was 12.5% higher than that of against HepG2/neo cell,while no increase in that of against HepG2/IL 12 cells. Cytotoxicities against K562 cells in HepG2/B7 1,HepG2/IL 12 groups were 19.38% and 14.78% higher than those of HepG2/neo group, but no significant difference between the first two groups. [WT5”HZ]Conclusion [WT8.75BZ]B7 1 and IL 12 gene transfer could remarkably promote immunogenicity of hepatocellular carcinoma cells and induce strong specific and nonspecific immunity against hepatocellular carcinoma in vitro.