肺腺癌A549细胞培养上清液对人单核细胞衍生树突状细胞的影响

目的：观察肺腺癌A549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A549免疫应答的影响。方法：制备人肺腺癌A549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A549免疫应答。结论：TSN能干扰MoDC成熟过程，下调基表型表达，诱导其凋亡发生、抑制其介导的抗肿瘤免疫应答；而脱离TSH影响、体外正常制备的MoDC瘤苗能够诱导出高铲的抗瘤免疫应答。

Effects of tumor supernatant of A549 lung adenocarcinoma on human monocyte-derived dendritic cells

Aim: To investigate the effects of tumor supernatant (TSN) of A549 lung adenocarcinoma on human monocyte derived dendritic cells (MoDC), and to study whether functional MoDC pulsed with A549 tumor antigen is effective in inducing antitumor immune response. Methods:Human TSN,inactivated TSN and tumor antigen were prepared. The 2 kinds of TSN were added to the MoDC culture medium throughout the whole culture course or only at the late stage (the 7th d)in 7 groups with different culture conditions; MoDC were pulsed with tumor antigen on the 4th d or not and were collected on the 9th d. MoDC phenotypes were analyzed by flow cytometry (FCM); the cell apoptosis was observed by PI staining and FCM; function was evaluated by mixed lymphocyte reaction (MLR),cytotoxic T lymphocyte (CTL) assay and IL 12 in the supernatant was detected by using ELISA. Results: When MoDC exposed to TSN in the whole culture course, phenotypes showed unobviously,the apoptotic cell ratio was significantly higher, MLR,CTL,and IL 12 decreased significantly. When MoDC exposed to TSN at the late culture stage or exposed to inactivated TSN throughout the whole culture course,no inhibition effects as above were demonstrated. When MoDC were free from TSN and pulsed with lung adenocarcinoma cell antigen, the phenotypes and function showed obviously. Conclusions: TSN could up regulate phenotypic maturation of MoDC, induce apoptosis and inhibit the antitumor immunity of mature MoDC. The normal MoDC pulsed with tumor antigen could induce higher antitumor immune response in vitro.