CAJ | 학술논문

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10 μg/ml Mtb P19作用于拂波醇酯(PMA)分化的THP-1细胞,CO_2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.
목적 탐토결핵분지간균19kD지단백(Mtb P19)대단핵거서세포표면TLR-2표체적영향.방법 이10 μg/ml Mtb P19작용우불파순지(PMA)분화적THP-1세포,CO_2부상부육6 h.면역세포화학법(ICC)관찰거서세포TLR-2분포;대거서세포TLR-2진행형광항체염색,류식세포술분석P19작용전후TLR-2적표체변화;격광공취초현미경관찰기수체배렬.결과 ICC법관찰발현TLR-2균균분포우거서세포표면.P19작용조여대조조상비평균형광강도명현증강;재작용조세포표면출현다개반괴상염색양성적형광표기구,이대조조TLR-2분자총체정현수궤동태적분포.결론 Mtb P19불부능유도거서세포TLR-2표체증가,환가이인기수체발생공능성취집.
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% CO_2.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.