抗STAT3 shRNA对树突状细胞成熟影响研究及其靶向防龋DNA疫苗构建  

Effect of Anti-STAT3 shRNA on Dendritic Cell Maturation and Corresponding Targeted Anti-caries DNA Vaccine Construction

作　　者：刘苗苗[1] 张艳[1] 张偲[1] 樊明文[1,2] 郭继华[1,2] 

机构地区：[1]武汉大学口腔医学院牙体牙髓科,湖北省口腔基础医学重点实验室-省部共建国家重点实验室培育基地口腔生物医学教育部重点实验室(武汉大学),湖北武汉430079 [2]武汉大学口腔医院牙体牙髓科,湖北武汉430079

出　　处：《口腔医学研究》2016年第3期234-238,共5页Journal of Oral Science Research

基　　金：国家自然科学基金资助项目(编号:81170955);中央高校基本科研业务费资助(编号:2042014kf0270)

摘　　要：目的:研究以慢病毒为载体的抗小鼠STAT3shRNA对树突状细胞(DC)成熟的影响,构建抗STAT3shRNA树突状细胞靶向防龋DNA疫苗。方法:制备抗小鼠STAT3shRNA慢病毒,体外转染树突状细胞系DC2.4,用Western杂交检测抑制STAT3表达的效果;用流式细胞术检测CD40、CD80和CD86表达;在抑制DC2.4的STAT3表达后,用LPS刺激,流式细胞术检测其对DCs成熟的影响;将针对STAT3的特异性shRNA整合入DC靶向防龋DNA疫苗pGJA-P/VAX,转染HEK-293细胞检测抑制STAT3表达效果。结果:抗STAT3shRNA慢病毒转染DC2.4后,可明显抑制STAT3表达,并增加DC表面标记物CD40、CD80和CD86的表达;用LPS刺激后,CD40、CD80和CD86表达进一步增强;抗STAT3shRNA DC靶向防龋DNA疫苗转染HEK-293细胞后可以抑制STAT3表达。结论:小鼠抗STAT3shRNA慢病毒可有效抑制DC2.4STAT3的表达,STAT3沉默后可促进DC2.4成熟。成功构建抗STAT3shRNA DC靶向防龋DNA疫苗。Objective：To investigate the effect of anti-STAT3 shRNA on dendritic cell maturation and to construct an anti-careis DNA vaccine carrying anti-STAT3 shRNA to enhance the efficacy of DNA vaccination.Methods：Dendritic cell line DC2.4was infected by lentivirus containing shRNA targeting mouse STAT3.Total cellular protein was collected to detect expression level of STAT3 by western blot.CD40,CD80 and CD86of DC2.4were analyzed by FACS.LPS was used after infection to stimulate DC maturation.Anti-careis DNA vaccine carrying anti-STAT3 shRNA was constructed by cloning shRNA fragment into DC target anti-caries DNA vaccine pGJA-P/VAX.The efficiency of its inhibition on STAT3 expression was confirmed in HEK-293 cells.Results：Expression of STAT3 decreased in DC2.4infected by lentivirus containing shRNA targeting STAT3.The expression levels of CD40,CD80 and CD86of DC2.4increased upon STAT3 knockdown,which were further up-regulated after the stimulation of LPS.Anti-caries DNA vaccine with anti-STAT3 shRNA could inhibit STAT3 expression.Conclusion：Lentiviral mediated shRNA interference targeting STAT3 could inhibit the STAT3 expression level of DC2.4and increase the expression of CD40,CD80 and CD86molecules and the maturation of dendritic cells.Anticaries DNA vaccine against STAT3 was constructed successfully.

关 键 词：STAT3 慢病毒 树突状细胞 SHRNA DNA疫苗 

分 类 号：R780.1[医药卫生—口腔医学]