摘要
目的观察索曲妥林(sotrastaurin或AEB071)对人外周血中单个核细胞(PBMCs)、纯化CD4^+T细胞和CD8^+T细胞分泌细胞因子的抑制作用及其机制。方法采集健康志愿者的静脉血,肝素抗凝,密度梯度离心法分离PBMCs后纯化CD4^+T和CD8^+T细胞,应用抗CD3抗体联合CD28抗体进行刺激,加或不加入AEB071培养后,检测细胞因子的产生、转录因子的表达和细胞的活化与增殖。结果索曲妥林处理不促进外周血T细胞的凋亡,而能抑制T细胞分泌细胞因子IFN-γ、TNF-α和IL-2,且呈时间和剂量依赖性;进一步研究发现,索曲妥林通过下调STAT-1和STAT-4的磷酸化以及T-bet的表达抑制细胞因子的产生。同时经研究发现,索曲妥林处理后抑制T细胞的活化和增殖。结论索曲妥林抑制T细胞转录因子的表达与磷酸化而抑制细胞因子的产生,从而抑制人体免疫功能。为此,进一步研究索曲妥林免疫抑制的机制,对索曲妥林在器官移植、银屑病和溃疡性结肠炎的治疗方面的应用,具有参考价值。
To investigate the inhibitory effect and mechanism of sotrastaurin on the production of cytokines,human PBMCs and purified CD4^+ or CD8^+T cells were stimulated with anti-CD3 plus anti-CD28 or with immobilized anti-CD3 and soluble anti-CD28 in the presence or absence of sotrastaurin. The levels of IFN-γ, TNF-α and IL-2 in the culture supernatants were detected by ELISA;the production of cytokines and transcription factors including T-bet, p-STAT-1 as well as p-STAT-4 were analyzed by FACS. Data showed that sotrastaurin lacked sufficient cytotoxicity toward T cells from healthy volunteers but exhibited significant antiproliferation. In addition, sotrastaurin had significant suppressive effects on the production of IFN-γ, TNF-α and IL-2 in a dosetime-dependent manner. Further study revealed that sotrastaurin down-regulated the phosphorylation of STAT4 and STAT1 and the expression of T-bet. All these results indicated that the main mechanism of sotrastaurin inhibiting the cytokine production is down-regulating the phosphorylation of STAT1 and STAT4 and the expression of T-bet.The further study on the mechanism of sotrastaurin immunosuppression is of great significance for its application in the treatment of organ transplantation, psoriasis and ulcerative colitis.
作者
金水平
吴琼丽
康双朋
杨滨燕
吴长有
JIN Shuiping;WU Qiongli;KANG Shuangpeng;YANG Binyan;WU Changyou(Institute of Immunology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2019年第6期461-470,477,共11页
Immunological Journal
基金
国家自然科学基金面上项目(31670900)
