初治肺结核患者外周血树突状细胞免疫功能的体外研究

Study of immune function of dendritic cells in peripheral blood of tuberculosis patients after initial treatment

目的研究初治肺结核患者外周血树突状细胞(DC)表面成熟标志的表达、分泌细胞因子以及对T淋巴细胞的刺激作用。方法收集初治肺结核患者68例,其中菌阴组35例,菌阳组33例。收集健康体检者40例作为对照组。分离外周血单个核细胞,诱导其分化成DC。流式细胞术检测各组DC的表面标志CD83和CD86的表达率;MTT法检测DC促进同种异体T淋巴细胞的增殖能力;ELISA法检测DC培养上清中白细胞介素(IL)-12、IL-10、干扰素(INF)-γ的含量。结果初治肺结核患者与对照组相比,DC表面标志CD83和CD86的表达量均显著降低(P<0.05),刺激淋巴细胞增殖的能力均显著降低(P<0.05),培养上清中IL-12的含量均显著增高(P<0.05),培养上清中IL-10、INF-γ的含量各组间差异无统计学意义(P>0.05)。结论初治肺结核患者外周血DC的成熟度显著下降,刺激混合淋巴细胞增殖的能力也显著降低,分泌IL-12的能力增强。

Objective To investigate expressions of surface maturation markers, secreting cytokines and the stimulat-ing effect on T lymphocytes in the dendritic cells （DC） from peripheral blood of tuberculosis （TB） patients after loading dose treatment. Methods TB patients who received initial treatment （n=68） were collected at the fifth hospital of Shijiazhuang from 2013 June to 2014 January. Base on clinical diagnosis and treatment guidelines of tuberculosis, they were divided into sputum smear-negative group （35 cases） and sputum smear-positive group（33 cases）. Forty cases of healthy adult were se-lected as control group. Mononuclear cells were isolated from peripheral blood and were cultured in medium to differentiate into DCs. Expression levels of CD83 and CD86 on DCs were examined by flow cytometry. The proliferation of allogeneic mixed lymphocyte stimulated by DCs was dectected using MTT assay. Contents of IL-12, IL-10 and INF-γin the cultural supernatant of DCs and blood serum from TB patients were detected by ELISA. Results Compared with controls ,the ex-pressions of CD83 and CD86 on DCs in TB patients after loading dose treatment decreased obviously（P〈0.05）, and the ability to stimulate the proliferation of lymphocytes reduced evidently（P〈0.05）. What’s more, IL-12 level increased mark-edly（P〈0.05）while IL-10 and INF-γlevels presented no significant difference among the three groups （P〉0.05）. Conclu-sion The expressions of maturation markers of DC cells of the peripheral blood in TB patients after initial treatment de-creased. The ability of stimulating mixed lymphocyte proliferation is also significantly reduced while secretion of IL-12 was enhanced.