摘要
目的研究R848对人自然杀伤细胞(NK)杀伤功能的作用及其机制。方法分离健康志愿者外周血单个核细胞(PBMC)、纯化NK细胞,加或不加R848进行培养。分别在0、2、6、24、48h收集培养细胞,流式细胞术检测R848对NK细胞表面活化分子CD25和CD69表达的影响;检测R848活化的NK细胞杀伤靶细胞K562的作用、通过检测颗粒酶A、B、穿孔素、TRAIL和NK细胞与靶细胞共孵育后CD107a/b的表达,探讨R848促进NK细胞的杀伤功能机制。结果 R848活化的NK细胞在培养24h时CD69和CD25分子的表达百分率和平均荧光密度达到峰值,随后活化分子的表达有所下降;R848活化的NK细胞对靶细胞的杀伤功能明显增强,TRAIL在不同NK细胞亚群的表达显著增加(P<0.05)。当NK细胞与K562共孵育后,R848活化的NK细胞表面CD107a/b的表达较未刺激条件明显增加。结论 R848可直接作用于NK细胞促进其杀伤功能,其作用机制与NK细胞活化后释放颗粒酶和穿孔素增加,并且TRAIL的表达增加有关。
This paper is aimed to study the effects of R848 on the cytotoxicity of human natural killer cells and its mechanism.We isolated peripheral blood mononuclear cell（PBMC） and purified NK cells from normal human peripheral blood,and then cultured them with R848.The cultured cells were harvested at different time points （0,2,6,24 and 48 hours） for follow tests.The expression of activating molecules such as CD69 and CD25 were detected by flow cytometry;the cytolytic activity and mechanism of R848-activated NK cells were evaluated by detecting the expression of granzymes,perforin,TRAIL,and CD107a/b.The results indicated that the percentage and MFI of the expression of CD69 and CD25 molecules on R848-activated human NK cells reached to peak levels after culturing for 24 hours and then decreased slightly;the cytotoxicity of R848 activated NK cells were enhanced significantly.After coincubation with the K562 target cells the degranulation of granzymes and perforin of R848 activated NK cells were more significant than those of the unstimulated cells.Therefore,the expression of CD107a/b and TRAIL was increased （P〈0.05）.This study provides valuable information that R848 can directly promote the cytotoxic activity of human NK cells by enhancing the degranulation of granzymes and perforin,and by up-regulating the expression of TRAIL.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2010年第8期657-660,共4页
Immunological Journal
基金
教育部博士学科点专项科研基金资助项目(20070558213
20090171120050新教师类)
广东省自然科学基金博士启动项目(8451008901000131)
广东省医学科研基金资助项目(A2009161)
