摘要
目的：研究人IFN－γ基因对胆管癌细胞株免疫原性的影响。方法：构建携带人IFN－γcDNA的逆转录病毒载体pZIP－IFN－γ，导入人胆管癌细胞系QBC939，运用WesternBlot法分析MHCI类分子的表达，体外混合淋巴细胞、肿瘤细胞培养（MLTR）和淋巴细胞杀伤试验研究导入IFN－γ基因的QBC939细胞免疫原性的变化。结果：G418阳性QBC939－IFN－γ细胞分泌IFN－γ活性为每48h0～9．5IU／106，SouthernBlot和SlotBlot证实了目的基因的整合及表达。WesternBlot证明MHCI类分子表达水平是未转基因亲本瘤细胞的1．78倍。MLTR证实该瘤苗能明显刺激胆管癌病人外周血淋巴细胞的增殖，并能增强HLAA2位点相匹配的胆管癌病人外周血淋巴细胞的杀伤活性。结论：IFN－γ基因修饰瘤苗能增强胆管癌细胞系QBC939的免疫原性。
Objective: To investigate the changes of the immunogenicity of the cholangiocarcinoma cellline QBC 939 transduced with human interferon-γ gene. Methods: Retroviral vector pZIP-IFN-γ carrying human IFE-γ cDNA was constructed and transduced to QBC 939 cells. The expression of MHC class I antigenand the changes of the immunogenicity of the transduced QBC 939 cells were studied with Western blot,mixed lymphocyte and tumor cell reaction (MLTR) and assay of cytotoxicity on lymphocytes respectively.Results: The IFN-γ activity secretea by the G418 positive QBC939-IFN-γ cells was 0~9. 51 IU/106 cells/48h. Southern blot and Slot blot confirmed the presence of the incorporation and expression of the target gene.The expression level of MHC class I antigen was 1. 78 tirnes higher in the transduced cells than in the untransduced ones. MLTR revealed marked proliferation of lymphocytes in the peripheral blood of patients withcholangiocarcinoma. Enhancement of lymphocytic lysis was found in HLA A2 matched patients. ConclUsion:The immunogenicity of QBC 939 cells was enhanced after the transduction interferon-γ gene.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
1999年第7期469-471,共3页
Journal of Third Military Medical University
关键词
基因治疗
干扰素-Γ
肿瘤疫苗
胆管肿瘤
cholangiocarcinoma cell line
gene therapy
interferon-γ
tumor vaccine
immunogenecity
