摘要
目的了解共刺激分子B7-H1在小鼠中枢神经系统小胶质细胞上的表达,及LPS刺激后的表达变化情况,探讨小胶质细胞在大脑炎症状态下的免疫功能变化。方法利用小鼠小胶质细胞株N9,以LPS刺激其活化,应用RT-PCR、定量PCR、流式细胞术、免疫组化检测B7-H1在刺激前后的表达变化规律;分离培养小鼠原代小胶质细胞并进行GSA-IB4染色鉴定,应用免疫组化检测原代小胶质细胞在LPS刺激前后的表达变化。结果①小鼠小胶质细胞株N9在未刺激状态下表达B7-H1的mRNA和蛋白分子;LPS刺激细胞活化后释放肿瘤坏死因子α,一氧化氮增加,同时B7-H1的mRNA和蛋白表达水平增加。②分离培养的小鼠原代小胶质细胞经GSA-IB4染色鉴定纯度在95%以上,免疫组化显示原代小胶质细胞组成性表达B7-H1,LPS刺激后表达明显上调。结论小鼠小胶质细胞株N9小胶质细胞组成性表达共刺激分子B7-H1,LPS刺激后表达上调,提示其参与了中枢神经系统免疫应答的调节。
Objective To observe the expression of the new co-stimulatory molecule B7-H1 in mouse microglial cells and its changes after LPS activation, for investigating the immune function of microglial cells in the CNS inflammation. Methods The microglial cell line N9 was stimulated with LPS, and then the expression of B7-H1 was detected by RT-PCR, real-time RT-PCR, flow cytometry, and immunohischemistry. After isolation and cultivation, the primary microglial cells was identify by GSA-IB4 staining and the B7-H1 expression change after LPS stimulation was found by using immunohischemistry. Results ①Resting N9 could express B7-H1 and the LPS stimulation could up-regulate the expression as well as accumulate the production of TNF-α and NO. ② The isolated primary microglia cells, whose purity was more than 95% by GSA-IB4 staining, were constitutively expressed B7-H1 which up-regulated signifjcantly after LPS stimulation. Conclusion The N9 and primary microglial cells could constitutively express co-stimulatory molecule B7-H1 and the expression can be increased significantly by LPS stimulation, which maybe play an important role in modulating the CNS immune response.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2006年第3期243-246,251,共5页
Immunological Journal
基金
国家自然科学基金海外青年学者合作研究基金(30228018)
