摘要
目的:研究表达ESAT-6-Ag85A(ES85A)融合基因的侵入型乳酸菌对小鼠体内免疫特性的影响。方法:将真核表达载体p Valac与ES85A连接构建真核表达质粒,并将其分别电转到重组乳酸菌NC8-pSIP-409和NC8-pSIP-409-FnBPA感受态中,构建双质粒的乳酸菌p Valac-ES85A/409和p Valac-ES85A/Fn BPA。将其免疫BALB/c小鼠,检测表达ES85A的侵入型乳酸菌对小鼠树突状细胞(DCs)亚群CD80和CD83的影响以及血清中细胞因子的表达水平。结果:Western blot和免疫荧光证明ES85A蛋白成功表达;与空载体组相比,在小鼠免疫表达ES85A的侵入型乳酸菌后,小鼠脾细胞中CD11^+CD80^+(P<0. 001)和CD11^+CD83^+(P<0. 01)以及血清中细胞因子IL-4(P<0. 05)的表达都有所增加。结论:表达ES85A的侵入型乳酸菌能促进小鼠体内DCs的分化和成熟以及提高血清中细胞因子IL-4的表达,为后期研制抗结核分枝杆菌的DNA疫苗制剂奠定了基础。
Objective:The influence of invasive lactic acid bacteria expressing ESAT-6-Ag85A(ES85A)on immune characteristics in mice model was explored.Methods:Eukaryotic expression vector pValac was connected with ES85A to acquire eukaryotic expression plasmid,which was transformed into NC8-pSIP-409 and NC8-pSIP-409-FnBPA by electroporation and then constructed double-plasmids lactic acid bacteria,namely pValac-ES85A/409 and pValac-ES85A/FnBPA.To detect the influence on dendritic cells(DCs)subsets CD80 and CD83 in mice model as well as expression levels of cytokines in serum,BALB/c were immunized with above strains.Results:The expression of ES85A protein was proved by Western blot and immunofluorescence;compared to control vector,expression of spleen cells CD11+CD80+(P<0.001)and CD11+CD83+(P<0.01)as well as cytokine IL-4(P<0.05)in serum all were improved after mice were immunized with invasive lactic acid bacteria expressing ES85A.Conclusion:Invasive lactic acid bacteria expressing ES85A could promote the differentiation and maturity of DCs and then expression of cytokine IL-4 in serum in mice model,which was a foundation for later DNA vaccine against mycobacterium tuberculosis.
作者
刘晶
张赞
刘洋
杨桂连
姜延龙
王春凤
LIU Jing;ZHANG Zan;LIU Yang;YANG Gui-Lian;JIANG Yan-Long;WANG Chun-Feng(College of Animal Science and Technology,Jilin Provincial Engineering Research Center of Animal Probiotics,Minstry of Education Laboratory of Animal Production and Quality Security,Jilin Agricultural University,Changchun 130118,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第1期10-14,共5页
Chinese Journal of Immunology
基金
国家重点研发计划项目(2017YFD0501000)
国家自然科学基金项目(31672528
31602092)
吉林省教育厅科学技术研究项目(JJKH20170316KJ)
吉林省科技发展计划项目(20160519011JH
20170204034NY
2018020104NY)
吉林省产业创新专项基金项目(2016C063)资助
