膀胱癌患者一氧化氮对外周血树突状细胞影响.docVIP

膀胱癌患者一氧化氮对外周血树突状细胞影响 　【关键词】 膀胱肿瘤 Effects of nitric oxide on peripheral blood dendritic cells in patients with bladder cancer 　　【Abstract】 AIM: To investigate the effects of nitric oxide (NO) on peripheral blood dendritic cells (DCs) in patients with bladder cancer. METHODS: DCs were obtained from peripheral blood mononuclear cells (PBMC) in patients with bladder cancer. The DCs were cultured in the presence of granulocyte/macrophage colony stimulating factor (1×106 U/L), interleukin4 (5×105 U/L), tumor necrosis factorα (105 U/L) and tumorassociated antigen (TAA) extracted from human bladder cancer cell line T24. The phenotypes (CD1a, CD80, CD86 and HLADR) were determined by flow cytometry. The proliferation of lymphocytes induced by DCs and the cytotoxicity of cytotoxic T lymphocytes (CTL) were assayed in the presence of NO. Bladder cancer cell line T24 was cultured as target cells and the cytotoxicity of CTL was determined by MTT assay. RESULTS: The proliferation of lymphocytes induced by DCs was inhibited by NO in a dosedependent manner (Plt;0.01). The NO resulted in an inhibition of the cytotoxicity of CTL against bladder cancer cell line T24 (Plt;0.01). The DC in vitro induced by TAA effectively resulted in the proliferation of lymphocytes ［proliferation rate: (74±6)%］ and the killing effect was significant on the bladder cancer cell line T24 by CTL ［killing rate: (54±3)%］. CONCLUSION: NO can inhibit the proliferation and the activation of lymphocytes induced by DCs and downregulate the antitumor cytotoxicity of CTL in patients with bladder cancer. 　　【Keywords】 nitric oxide; dendritic cell; cytokine; bladder neoplasms 　　【摘要】 目的: 观察外源性一氧化氮（nitric oxide, NO）对膀胱癌患者外周血树突状细胞(dendritic cell, DC)的影响. 方法: 膀胱癌患者外周血中分离出单个核细胞，用粒/巨噬细胞集落刺激因子（GMCSF）、白细胞介素4（IL4）、肿瘤坏死因子（TNF）α和肿瘤相关抗原（tumor associated antigen, TAA）培养DC细胞. 外源性NO干预后DC刺激自体淋巴细胞的增殖和细胞毒性T细胞(CTL)对膀胱癌T24的杀伤效应，用MTT法检测. 结果: 外源性NO可明显抑制DC刺激的自体淋巴细胞增殖，在NO浓度分别为50， 100， 200和400 nmol/L时淋巴细胞增殖率分别为(67±3)%，(40±6)%，(24±2)%和(18±3)%（与