活动性肺结核患者单核来源巨噬细胞极化能力的研究

The research of polarization of monocyte-derived macrophages from patients with active tuberculosis

目的：研究活动性肺结核患者单核来源巨噬细胞（monocyte-derived macrophages, MDM）向 M1型和 M2型巨噬细胞极化的能力。方法采集活动性肺结核患者和健康人外周血,分离外周血单个核细胞（peripheral blood mononuclear cells,PBMCs）,采用磁珠分选技术纯化 CD14＋单核细胞,体外分化为 M0型巨噬细胞,然后分别用细菌脂多糖（lipopolysaccharide,LPS）/干扰素-γ（interferon-γ,IFN-γ）和 IL-4刺激巨噬细胞向 M1和 M2型巨噬细胞极化,最后流式细胞术检测 M1型 MDM 表面 CD80、CD86和 HLA-DR 的表达,M2型 MDM 表面 CD163和 CD206的表达。结果与健康人相比,活动性肺结核患者 M1型 MDM 表面 CD80和 CD86的表达显著降低（t =2.359,P 〈0.05；t =2.816,P 〈0.01）,而 M2型 MDM 表面 CD163和 CD206的表达显著升高（t =2.368,P 〈0.05；t =2.994,P 〈0.01）。结论活动性肺结核患者 MDM 向 M1型巨噬细胞极化能力减弱,而向M2型巨噬细胞极化能力增强。

Objective To study the polarization of monocyte-derived macrophages （MDMs） to the M1 and M2 macrophages,which were obtained from active pulmonary tuberculosis patients.Methods Peripheral bleed were collected from patients with active pulmonary tuberculosis and healthy people. Peripheral blood mononuclear cells （PBMCs） were separated from peripheral bleed.CD14 ＋ mononuclear cells were purified from PBMCs by magnetic activated cell sorting （MACS） and in vitro differentiated to M0 macrophages.Bacterial lipid polysaccharide （LPS）/interferon-γ（IFN-γ） and interleukin-4 （IL-4） were added to the culture of M0 macrophages and were used to stimulate the M0 macrophages to M1 and M2 macrophages,Flow cytometry method was used to detect the markers on/in M1 and M2 macrophages.The markers of CD80,CD86 and HLA-DR were mainly expressed on/in M1 macrophages and the markers of CD1 63 and CD206 were mainly expressed on/in M2 macrophages.Results Compared with healthy people,CD80 and CD86 expressed in M1 macrophages collected from patients with active pulmonary tuberculosis were significantly decreased （t = 2.359,P 〈0.05;t = 2.81 6,P 〈0.01）,and CD1 63 and cd206 expressed in M2 macrophages surface were significantly increased （t = 2.368,P 〈0.05;t =2.994,P 〈0.01）.Conclusions Polarization ability of MDM to M1 macrophages is decreased and to M2 macrophages are enhanced,which were collected from patients with active pulmonary tuberculosis.