活动性结核患者单核来源巨噬细胞C-C基序配体5的表达研究

The study of the C-C motif ligand 5 expression in monocyte-derived macrophage from active tuberculosis patients

目的 研究活动性结核患者单核来源巨噬细胞（MDM）趋化因子C-C基序配体5（CCL5）的表达水平.方法 收集309医院的活动性肺结核患者和健康人抗凝血,分离纯化单核细胞并体外培养使其分化为初始型（M0）巨噬细胞.然后分别用细菌脂多糖（LPS）/γ-干扰素（IFN-y）和白细胞介素4刺激24h,使其向促炎症型（M1）巨噬细胞和抗炎症（M2）型巨噬细胞极化,收集细胞并提取总RNA,荧光定量PCR检测CCL5 mRNA的表达.结果 活动性结核患者M0、M1和M2型MDM中CCL5的相对表达量分别为（0.023 ±0.012）、（0.675±0.337）和（0.037 ±0.031）,健康人M0、M1和M2型MDM中CCL5的相对表达量分别为（0.051 ±0.026）、（0.727±0.376）和（0.068 ±0.045）.与健康人相比,活动性结核患者M0和M2型MDM中CCL5的表达显著降低（U=52.5,P＜0.001;t=2.336,P＜0.05）,而M1型MDM中CCL5的表达没有显著变化（t=0.4307,P＞0.05）.结论 活动性结核患者MDM细胞中CCL5的表达降低,提示巨噬细胞CCL5参与结核病的感染免疫.

Objective To study the expression of C-C motif ligand 5 （CCL5） on monocyte-derived macrophage from active tuberculosis patients.Methods Patients with active pulmonary tuberculosis and healthy subjects were recruited from 309 hospital.Monocytes were isolated from peripheral blood and cultured in vitro for differentiation to macrophages.Macrophages were then activated with LPS/IFN-γor IL-4 for 24 hours to polarize to M1 type macrophages or M2 type macrophages.Total RNA was extracted from macrophages.Fluorescence quantitative PCR was used to detect the mRNA expression of CCL5.Results The expression of CCL5 mRNA in M0 type MDM from patients with active pulmonary tuberculosis was significantly lower than that of healthy controls[（0.023 ± 0.012） vs （0.051 ± 0.026）,U =52.5,P 〈 0.001].Similarly,the expression of CCL5 mRNA in M2 type MDM from active tuberculosis patients was significantly lower than that of healthy controls [（0.037 ± 0.031） vs （0.068 ± 0.045）,t =2.336,P 〈 0.05].However,the expression of CCL5 mRNA in M1 type MDM from active tuberculosis patients was similar with that of healthy controls[（0.675 ± 0.337）vs （0.727 ± 0.376）,t =0.4307,P 〉 0.05）.Conclusion The expression of CCL5 in M0 and M2 type MDM from patients with active tuberculosis was decreased,suggesting that the CCL5 of macrophages participates in the anti-tuberculosis immunity.