目的研究阻断膀胱肿瘤患者外周血树突状细胞(dendriticcell,DC)PD—L1通路对其免疫功能的影响。方法通过rhGM.CSF、rhIL-4在体外诱导膀胱肿瘤患者和正常人外周血DC,PD—Ll单克隆抗体处理后,流式细胞仪检测CD1a HLA和CD83的表达;噻唑盐比色法和酶联免疫吸附法检测其刺激淋巴细胞增殖和分泌IL-10和IL—12能力的变化。结果阻断PD—L1通路对DC成熟无明显影响,但可以使膀胱肿瘤患者DC刺激淋巴细胞增殖能力增强(4.004-1.28和1.494-0.45),并增强IL-12[(108.30±21.89)ng/L和(37.17±14.89)ng/L]、减少IL-10[(108.904-21.77)ng/L和(214.994-54.99)ng/L]的分泌(P〈0.05)。结论阻断PD—L1通路可使膀胱肿瘤患者外周血DC的免疫活性增强,提示可尝试通过PD—L1通路调节膀胱肿瘤患者的免疫功能,为防治膀胱肿瘤复发提供新途径。
Objective To investigate the immunological function variation of peripheral dendritic cell (DC) from bladder cancer patients after blocking PD-L1 pathway. Methods DC from normal control and patient with bladder cancer were cultured with rhGM-CSF, rhIL-4 in vitro and treated with PD-L1 mono- clonal antibody. The expression of CDla, HLA and CD83 were examined by flow cell meter. The effect of DC induced lymphatic cellular proliferation and its capability of secreting IL-10, IL-12 were determined by MTT and ELISA. Results Blocking PD-LI pathway did not infuluence the maturation of DC. But the DCs from bladder cancer patient signifcantly boosted the lymphatic cellular proliferating (4.00 ± 1.28 versus 1.49 ± 0.45) and IL-12 secretion capability (108.30 ± 21.89 versus 37. 17 ± 14.89 ng/L) , and yet it also de- creased the secretion of IL-10 ( 108.90 ± 21.77 versus 14.99 ± 54.99 ng/L) after blocking PD-L1 pathway (P 〈 0.05). Conclusion Blocking PD-LI pathway on the DC from bladder cancer patient may improve its anticancer immunological function.