【摘要】 目的探讨结核分枝杆菌19kD脂蛋白(Mtb P19)对单核巨噬细胞表面TLR-2表达的影响。方法以10μg/ml Mtb P19作用于拂波醇酯(PMA)分化的THP-1细胞,CO2孵箱孵育6 h。免疫细胞化学法(ICC)观察巨噬细胞TLR-2分布;对巨噬细胞TLR-2进行荧光抗体染色,流式细胞术分析P19作用前后TLR-2的表达变化;激光共聚焦显微镜观察其受体排列。结果ICC法观察发现TLR-2均匀分布于巨噬细胞表面。P19作用组与对照组相比平均荧光强度明显增强;在作用组细胞表面出现多个斑块状染色阳性的荧光标记区,而对照组TLR-2分子总体呈现随机动态的分布。结论Mtb P19不仅能诱导巨噬细胞TLR-2表达增加,还可以引起受体发生功能性聚集。更多还原

【Abstract】 To observe the effect of Mycobacterium tuberculosis 19 kDa lipoprotein(Mtb P19) upon the expression and distribution of Toll-like receptor-2(TLR-2) on the surface of macrophages,Mtb P19 was prepared from the cultured M.tuberculosis H37 Ra strain,and phorbol myristatye acetate(PMA)-differentiated THP-1 cells were co-cultivated with Mtb P19 at concentration of 10 g/mL and at 37 ℃ temperature and a condition containing 5% CO2.for 6 hours..The distribution of TLP-2 on the surface of macrophages was investigated by immuno-cellular chemical method(ICC) while the effect of Mtb P19 upon the expression of TLR on the surface of macrophages was assayed by fluorescent antibody staining.In addition,the changes of TLR-2 expression before and after the effect of Mtb P19 were investigated by flow cytometry analysis and change of TLR-2 arrangement after stimulation with Mth P19 was determined by co-focal microscopy.It was found that the TLR-2 molecules were evenly distributed on the surface of macrophages as demonstrated by ICC.The mean fluorescent intensity increased significantly after stimulation with Mtb P19 for 6 hours in comparison with that of the control group,and the patchy surface with fluorescent staining positive zones could be detected on the surface of macrophages.Nevertheless,the distribution of TLR-2 molecule in the control group appeared to be randomly dynamic.It is evident that the Mth P19 not only can induce the surface expression of TLR-2 molecules,but also cause a functional aggregation of this receptor.更多还原