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目的 :观察肺腺癌A5 4 9细胞培养上清液 (TSN)对人单核细胞衍生树突状细胞 (MoDC)的表型、增殖和抗肿瘤A5 4 9免疫应答的影响。方法 :制备人肺腺癌A5 4 9细胞的TSN、灭活TSN和冻融抗原 ;人MoDC培养按照不同培养条件组合分为 7组 ,分别于培养全程或晚期 (第 7d)加入TSN或第 4d加入冻融抗原冲击致敏 ,制备MoDC瘤苗 ,第 9d收获细胞。检测各组MoDC表型、MoDC凋亡率、体外混合淋巴细胞反应 (MLR)、MoDC诱导的细胞毒性T细胞 (CTL)抑瘤活性、培养上清液中IL 12含量。结果 :培养全程加TSN的MoDC ,相对特异性表型CD80、CD83和人类白细胞抗原 (HLA DR)明显下调 ,凋亡率明显升高 ,同时 ,激活T细胞增殖能力、诱导CTL抑瘤活性及分泌IL 12的量均显著低于正常培养组 ;培养晚期加TSN的MoDC及培养全程加入灭活TSN的MoDC ,均无上述影响 ;脱离TSN影响、体外正常制备的MoDC瘤苗 ,表型明显上调、各项免疫指标明显升高 ,诱导出了高效的抗A5 4 9免疫应答。结论 :TSN能干扰MoDC成熟过程 ,下调其表型表达 ,诱导其凋亡发生 ,抑制其介导的抗肿瘤免疫应答 ;而脱离TSN影响、体外正常制备的MoDC瘤苗能够诱导出高效的抗瘤免疫应答
Objective: To observe the effect of TSN on the phenotype, proliferation and anti-tumor A549 immune response of human monocyte-derived dendritic cells (MoDC). Methods: TSN, inactivated TSN and freeze-thaw antigen of human lung adenocarcinoma A549 cells were prepared. Human MoDC was divided into 7 groups according to different culture conditions. TSN or 4d Add freeze-thaw antigen impact sensitization, preparation MoDC tumor vaccine, cells were harvested on the 9th day. MoDC phenotype, MoDC apoptosis rate, mixed lymphocyte reaction (MLR), MoDC-induced cytotoxic T lymphocyte (CTL) anti-tumor activity and IL 12 content in culture supernatant were detected. Results: MoDC with TSN increased significantly, and the relative specific phenotypes of CD80, CD83 and HLA - DR significantly decreased, and the apoptosis rate was significantly increased. At the same time, the proliferation of T cells was stimulated and the antitumor activity and secretion of CTL were induced The amount of IL-12 was significantly lower than that of the normal culture group. MoDC with TSN cultured in the late stage and MoDC with inactivated TSN were not affected by TSN. Various immune indicators were significantly increased, induced a highly efficient anti-A549 immune response. CONCLUSION: TSN can interfere with the maturation of MoDC, down-regulate its phenotype and induce its apoptosis, and inhibit its anti-tumor immune response. However, the TSN-induced MoDC vaccine can induce highly effective anti-tumor Immune response