摘要
目的探讨细胞因子信号传导抑制因子-1(SOCS1)基因腺病毒转染对未成熟的树突状细胞(DC)的成熟及细胞因子分泌功能的影响。方法构建SOCS1腺病毒并PCR鉴定,感染体外分离、培养的小鼠骨髓DC(阳性转染组),Western blot法检测SOCS1的表达,以阴性转染组(转染Ad-GFP)及空白对照组(未经病毒转染)为对照,采用脂多糖(LPS)刺激诱导各组DC成熟,通过流式细胞术测定LPS刺激前后各组DC表面CD40及CD80的表达水平,ELISA法检测细胞因子白细胞介素(IL)-10及IL-12的分泌水平。结果成功构建并鉴定转染腺病毒,感染DC后,阳性转染组SOCS1蛋白表达水平较阴性转染组及空白对照组显著增高。LPS刺激后,阳性转染组CD40及CD80的表达较阴性转染组及空白对照组显著降低(P<0.05),IL-12分泌水平明显较低(P<0.05),而IL-10分泌水平显著增高(P<0.05)。结论通过腺病毒转染使SOCS1在DC中过度表达,不仅能有效抑制DC的成熟,还能抑制LPS介导的DC分泌细胞因子IL-12,并诱导抑制性细胞因子IL-10的分泌。
Objective To investigate the effect of suppressor of cytokine signaling - 1 （SOCS1） by transfection of adenovirus vectors on the phenotype and immunologic function of dendritic cells. Methods The adenovirus vectors con- taining SOCS1 was constructed and confirmed by PCR. The phenotype CD40 and CDS0, and cytokine secretion of deudefi- tic cells was measured after LPS stimulation by flow cytometry and ELISA methods. Results The expression of SOCS1 in dendritic cells transfected by adenovirus vectors （ DC - SOCS1 ） was efficient. DC - SOCS1 showed significantly lower ex- pressions of CD40 and CD80, and reduced IL - 12 despite of the stimulation of LPS （ P 〈 0. 05 ）. Furthermore, signifi- cantly higher IL - 10 was found in the DC - SOCS1 compared to control group （ P 〈 O. 05）. Conclusion SOCS1 expres- sion could block maturation and secretion of inflammatory cytokine IL - 12, and promote production of inhibitory cytokine IL - 10 induced by LPS in DC.
出处
《广东医学》
CAS
CSCD
北大核心
2013年第2期169-172,共4页
Guangdong Medical Journal
基金
广东省科技计划项目(编号:2010B031600014)
