DEN2对鼠源性DCTLR7、MyD88、NF-κB的表达与细胞因子分泌的影响

发布时间：2018-01-03 13:03

本文关键词：DEN2对鼠源性DCTLR7、MyD88、NF-κB的表达与细胞因子分泌的影响　出处：《中国免疫学杂志》2013年06期 　论文类型：期刊论文

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【摘要】：目的:观察登革2型病毒(DEN2)感染鼠源性DC后TLR7、MyD88、NF-κB的表达及IFN-α、IP-10的分泌变化,探讨MyD88依赖型信号通路在DEN2感染中的作用。方法:rmGM-CSF和rmIL-4联合诱导C57BL/6小鼠骨髓源性树突状细胞(BMDC),常规方法增殖鉴定DEN2,利用DEN2(MOI=0.4)感染BMDC,直接免疫荧光检测DEN2吸附鼠源性BMDC;West-ern blot检测感染后不同时间TLR7、MyD88、NF-κB蛋白表达。双抗体夹心ELISA法检测感染后不同时间细胞培养上清IP-10、IFN-α的水平,RT-PCR检测相应时间点被感染DC内DEN2 NS5核酸水平。结果:rmGM-CSF和rmIL-4联合诱导C57BL/6小鼠BMDC,5天后获得纯度为70%的相对未成熟DC;与对照组相比DEN2感染BMDC后24小时TLR7、MyD88、NF-κB的表达升高;48小时TLR7表达低于正常对照组,但MyD88、NF-κB的表达高于正常对照组;72小时DEN2感染组TLR7、MyD88、NF-κB的表达均较前降低,且TLR7、MyD88的表达低于正常对照组。DEN2感染组细胞培养上清IFN-α、IP-10的水平明显高于正常对照组,IFN-α逐渐升高,72小时分泌量达最高,可分泌(933.94±29.02)ρg/ml;IP-10在48小时分泌达高峰,分泌量为(834.44±43.60)ρg/ml,结果具有统计学意义(P0.05),且被感染DC内DEN2 NS5病毒核酸水平48小时达最高,72小时有所降低。结论:DEN2可影响小鼠BMDC TLR7、MyD88、NF-κB的表达,促进其分泌IFN-α、IP-10进而影响病毒复制。
[Abstract]:Objective: to observe the expression of NF- 魏 B and the secretion of IFN- 伪 -mil IP-10 in murine DC infected with dengue type 2 virus (DEN2). To investigate the role of MyD88 dependent signaling pathway in DEN2 infection methods the bone marrow-derived dendritic cells (BMSCs) of C57BL / 6 mice were induced by 1: rmGM-CSF and rmIL-4. BMDC. DEN2 was detected by routine method. BMDCwas infected with DEN2MoI0. The DEN2 adsorbed murine BMDCwas detected by direct immunofluorescence. West-ern blot was used to detect MyD88 at different time after infection. The expression of NF- 魏 B protein. The level of IP-10 IFN- 伪 in the supernatant of cell culture at different time after infection was detected by double antibody sandwich ELISA method. RT-PCR was used to detect the level of DEN2 NS5 nucleic acid in the infected DC at the corresponding time point. Results the BMDC of C57BL / 6 mice was induced by the combination of rmIL-4 and 7% rmGM-CSF. After 5 days, relative immature DCS with purity of 70% was obtained. Compared with the control group, the expression of NF- 魏 B increased 24 hours after DEN2 infection with BMDC. At 48 hours, the expression of TLR7 was lower than that of normal control group, but the expression of MyD88NF-kappa B was higher than that of normal control group. The expression of NF- 魏 B in TLR7, MyD88 and NF-kappa B in 72 hours DEN2 infected group was lower than that in the former group, and TLR7. The expression of MyD88 was lower than that of normal control group. The level of IFN- 伪 IP-10 in the culture supernatant of DEN2 infected group was significantly higher than that of normal control group. At 72 hours, the secretion reached the highest level, which could secrete 933.94 卤29.02 蟻 g / ml; The peak secretion of IP-10 was 834.44 卤43.60 蟻 g / ml at 48 h. The results were statistically significant (P 0.05). And the nucleic acid level of DEN2 NS5 virus in the infected DC reached the highest level at 48 hours to 72 hours. Conclusion: 1 DEN2 can affect BMDC TLR7 / MyD88 in mice. The expression of NF- 魏 B stimulates the secretion of IFN- 伪 -IP-10 and further affects viral replication.
【作者单位】：贵阳医学院免疫学教研室;
【基金】：国家自然科学基金资助项目(31060129) 贵州省优秀人才省长基金贵州省教育厅“125”重大科技专项资助
【分类号】：R392.12
【正文快照】： 登革病毒(Dengue virus,DEN)主要由埃及伊蚊及白纹伊蚊传播,其感染已成为全球性重要的公共卫生问题,其中以登革2型病毒(Dengue virustype 2,DEN2)感染最为常见和严重[1]。目前认为登革病毒的致病机制主要与机体的免疫功能和病毒毒力有关,由此提出的发病机制包括抗体依赖的增

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