本文關(guān)鍵詞：DEN2對(duì)鼠源性DCTLR7、MyD88、NF-κB的表達(dá)與細(xì)胞因子分泌的影響　出處：《中國(guó)免疫學(xué)雜志》2013年06期 　論文類型：期刊論文

  更多相關(guān)文章： DEN 樹(shù)突狀細(xì)胞 MyD 細(xì)胞因子

【摘要】：目的:觀察登革2型病毒(DEN2)感染鼠源性DC后TLR7、MyD88、NF-κB的表達(dá)及IFN-α、IP-10的分泌變化,探討MyD88依賴型信號(hào)通路在DEN2感染中的作用。方法:rmGM-CSF和rmIL-4聯(lián)合誘導(dǎo)C57BL/6小鼠骨髓源性樹(shù)突狀細(xì)胞(BMDC),常規(guī)方法增殖鑒定DEN2,利用DEN2(MOI=0.4)感染BMDC,直接免疫熒光檢測(cè)DEN2吸附鼠源性BMDC;West-ern blot檢測(cè)感染后不同時(shí)間TLR7、MyD88、NF-κB蛋白表達(dá)。雙抗體夾心ELISA法檢測(cè)感染后不同時(shí)間細(xì)胞培養(yǎng)上清IP-10、IFN-α的水平,RT-PCR檢測(cè)相應(yīng)時(shí)間點(diǎn)被感染DC內(nèi)DEN2 NS5核酸水平。結(jié)果:rmGM-CSF和rmIL-4聯(lián)合誘導(dǎo)C57BL/6小鼠BMDC,5天后獲得純度為70%的相對(duì)未成熟DC;與對(duì)照組相比DEN2感染BMDC后24小時(shí)TLR7、MyD88、NF-κB的表達(dá)升高;48小時(shí)TLR7表達(dá)低于正常對(duì)照組,但MyD88、NF-κB的表達(dá)高于正常對(duì)照組;72小時(shí)DEN2感染組TLR7、MyD88、NF-κB的表達(dá)均較前降低,且TLR7、MyD88的表達(dá)低于正常對(duì)照組。DEN2感染組細(xì)胞培養(yǎng)上清IFN-α、IP-10的水平明顯高于正常對(duì)照組,IFN-α逐漸升高,72小時(shí)分泌量達(dá)最高,可分泌(933.94±29.02)ρg/ml;IP-10在48小時(shí)分泌達(dá)高峰,分泌量為(834.44±43.60)ρg/ml,結(jié)果具有統(tǒng)計(jì)學(xué)意義(P0.05),且被感染DC內(nèi)DEN2 NS5病毒核酸水平48小時(shí)達(dá)最高,72小時(shí)有所降低。結(jié)論:DEN2可影響小鼠BMDC TLR7、MyD88、NF-κB的表達(dá),促進(jìn)其分泌IFN-α、IP-10進(jìn)而影響病毒復(fù)制。
[Abstract]:Objective: to observe the expression of NF- 魏 B and the secretion of IFN- 偽 -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- 偽 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- 偽 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 鹵29.02 蟻 g / ml; The peak secretion of IP-10 was 834.44 鹵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- 偽 -IP-10 and further affects viral replication.
【作者單位】： 貴陽(yáng)醫(yī)學(xué)院免疫學(xué)教研室;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(31060129) 貴州省優(yōu)秀人才省長(zhǎng)基金 貴州省教育廳“125”重大科技專項(xiàng)資助
【分類號(hào)】：R392.12
【正文快照】： 登革病毒(Dengue virus,DEN)主要由埃及伊蚊及白紋伊蚊傳播,其感染已成為全球性重要的公共衛(wèi)生問(wèn)題,其中以登革2型病毒(Dengue virustype 2,DEN2)感染最為常見(jiàn)和嚴(yán)重[1]。目前認(rèn)為登革病毒的致病機(jī)制主要與機(jī)體的免疫功能和病毒毒力有關(guān),由此提出的發(fā)病機(jī)制包括抗體依賴的增

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本文編號(hào)：1373989