外源性IFN-β对OL细胞和OL/BDV细胞中内源性Ⅰ型IFN的诱导作用  

Effect of exogenous IFN-βon the induction of endogenous typeⅠIFN in OL and OL/BDV cells

作　　者：翟爱霞[1] 颜冬梅[2] 王海萍[2] 崔乐乐 卜桐 考文萍[1] 张凤民[1] Zhai Aixia;Yan Dongmei;Wang Haiping;Cui Lele;Bu Tong;Kao Wenping;Zhang Fengmin(Department of Microbiology,Harbin Medical University Wu Lien-Teh institute,Heilongjiang Provincial Key Laboratory of Infection and Immunity Key Laboratory of Pathogen Biology in Heilongjiang Provincial Education Institute Science and Technology Innovation Team for Infection and Immunity in Heilongjiang Provincial Education Institute,Harbin 150081,China;The Fourth Affiliated Hospital of Harbin Medical University 150001,China)

机构地区：[1]哈尔滨医科大学微生物学教研室伍连德研究所、黑龙江省感染与免疫重点实验室、黑龙江省普通高校病原生物学重点实验室、黑龙江省普通高校感染与免疫科技创新团队,哈尔滨150081 [2]哈尔滨医科大学附属第四医院,150001

出　　处：《国际免疫学杂志》2019年第1期1-6,共6页International Journal of Immunology

基　　金：国家自然科学基金(81501737);黑龙江省教育厅科学技术研究项目(12541365).

摘　　要：目的探讨外源性干扰素(interferon,IFN)-β对人类少突胶质细胞(oligodendrocyte,OL)和博尔纳病病毒(Borna disease virus,BDV)持续感染的OL细胞(OL/BDV)中内源性Ⅰ型IFN表达和干扰素调节因子(interferon regulatory factor,IRF)7细胞定位的影响。方法外源性IFN-β处理OL细胞和OL/BDV细胞0、0.5、2、4、8、24 h,根据细胞是否加入外源性IFN-β,分为外源性IFN-β未刺激组和刺激组。采用实时荧光定量PCR(quantitative real-time PCR,qPCR)检测Ⅰ型IFN mRNA表达。OL细胞和OL/BDV细胞分别转染IRF7-EGFP质粒,外源性IFN-β刺激4 h,观察IRF7细胞定位情况。结果外源性IFN-β刺激OL细胞,在2、4、24 h,IFN-αmRNA表达显著增加(P值分别为0.008,0.0001,0.004);在0.5、4、8、24 h,IFN-βmRNA表达显著增加(P值分别为0.0004,0.0002,0.011,0.012),两者均在4 h增加最为显著。外源性IFN-β刺激OL/BDV细胞4 h和24 h,IFN-αmRNA表达显著增加(P值分别为0.0004,0.022),IFN-βmRNA表达仅在刺激4增加(P=0.002)。外源性IFN-β刺激OL细胞和OL/BDV细胞4 h,IRF7绿色荧光蛋白均转移至细胞核内。结论外源性IFN-β可诱导OL细胞和OL/BDV细胞中内源性I型IFN表达,参与解除BDV对I型IFN应答的抑制过程,并促进IRF7的活化入核。Objective To investigate the effect of exogenous interferon (IFN) -β on the expression of endogenous typeⅠIFN and the localization of interferon regulatory factor (IRF) 7 in human oligodendrocyte (OL) and Borna disease virus (BDV) persistently infected OL cells (OL/BDV). Methods The expression of type Ⅰ IFN mRNA was detected by quantitative real-time PCR (qPCR) after OL cells and OL/BDV cells were treated with exogenous IFN-β for 0,0.5,2,4,8 h and 24 h.According to whether the exogenous IFN-β was added to the cell culture,cells were divided into exogenous IFN-β unstimulated and stimulated cells.The localization of IRF7 was observed after OL cells and OL/BDV cells were transfected with IRF7-EGFP plasmids and stimulated with exogenous IFN-β for 4 h. Results The expression of IFN-α mRNA was increased when OL cells were stimulated by exogenous IFN-β for 2,4 h,and 24 h (P values were 0.008,0.0001,0.004,respectively).The expression of IFN-β mRNA was increased when cells were treated for 0.5,4,8,24 h(P values were 0.0004,0.0002,0.011,0.012 respectively).The most significant increase for IFN-α and IFN-β mRNA were all at 4 h after treatment.The expression of IFN-α mRNA in OL/BDV cells which were treated with the same stimulation as for OL cells,increased at 4 h and 24 h(P values were 0.0004,0.022 respectively).The expression of IFN-β mRNA increased only when treated for 4 h (P=0.002).Meanwhile,IRF7 green fluorescent protein translocated into the nucleus in OL and OL/BDV cells treated with exogenous IFN-β stimulated for 4 h. Conclusions Exogenous IFN-β induces the expression of endogenous typeⅠIFN in OL and OL/BDV cells.Moreover,endogenous IFN-β participates in releasing the suppression of BDV toward typeⅠIFN,and promotes the activation of IRF7.

关 键 词：外源性干扰素-β 人类少突胶质细胞 博尔纳病病毒持续感染的人类少突胶质细胞 内源性I型干扰素 

分 类 号：R373[医药卫生—病原生物学]