结核分枝杆菌P19对巨噬细胞功能及表型影响的研究

Influence of Mycobacterium tuberculosis 19-kDa lipoprotein on function and phenotype of macrophage

目的探讨结核分枝杆菌19kD脂蛋白(MtbP19)对单核巨噬细胞功能及表型的影响。方法以10μg/mLMtbP19作用于拂波醇酯(PMA)分化的THP-1细胞,CO2孵箱孵育,酶联免疫吸附法(ELISA)检测不同时间巨噬细胞TNF-α和IL-6的分泌水平;对巨噬细胞表型分子HLA-DR进行荧光抗体染色,流式细胞术分析P19对巨噬细胞表型的影响;流式细胞术及光学显微镜研究MtbP19对巨噬细胞吞噬活性的影响。结果P19能够诱导巨噬细胞TNF-α和IL-6分泌水平的增加,呈时间依赖关系;经P19刺激的巨噬细胞HLA-DR表达增强,平均荧光强度(MFI)为(54.3±3.4)%,与BSA对照组(34.3±3.6)%比较P<0.05;P19能增强巨噬细胞的吞噬活性。结论MtbP19能诱导巨噬细胞活化,在感染过程中可能具有一定的抗结核保护作用。

To observe the influence of Mycobacterium tuberculosis 19-kDa lipoprotein（Mtb P19） on function and phenotype of macrophage,the Mtb P19 was prepared from cultured Mtb H37Ra and the phorbol myristate acetate-differentiated THP-1 cells were incubated with P19 at the concentration of 10 μg/mL with 5% CO2 at 37℃ for up to 48 hours.Supernatants were collected for TNF-α and IL-6 detection by ELISA,then the phenotype fluorescent antibodies were stained to analyze HLA-DR expression changes between control group and experimental group.Flow cytometry and microscopy was used to assay the phagocytosis in macrophages stimulated by Mtb P19.IL-6 and TNF-α in collected supernatants were detected.Results indicated that both were found significant increases and their phagocytosis were enhanced.Comparing to the control group,the mean fluorescence intensity showed a significant increase 24hs after stimulation.It presents that Mtb P19 could be able to induce macrophages activation,and it would be significantly important for protection during infection period.