本文選題：細(xì)粒棘球蚴 + Eg10��； 參考：《寧夏醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的初步明確細(xì)粒棘球蚴重組抗原Eg10和m MDH免疫小鼠產(chǎn)生的免疫耐受機(jī)制。方法1.體外培養(yǎng)小鼠骨髓來(lái)源的樹突狀細(xì)胞(Dendritic cell,DC),用Eg10和m MDH體外刺激DCs,通過(guò)掃描電鏡觀察DCs的形態(tài)變化,細(xì)胞免疫熒光觀察DCs表達(dá)吲哚胺2,3雙加氧酶(Indoleamine 2,3-dioxygenase,IDO)的水平;2.利用IDO抑制劑1-甲基色氨酸(1-Methyl-tryptophan,1-MT)處理DCs 3h,用來(lái)抑制IDO的表達(dá)及酶活性,然后用重組抗原Eg10和m MDH體外刺激DCs,通過(guò)混合淋巴細(xì)胞反應(yīng)檢測(cè)DCs刺激CD4+T細(xì)胞增殖及誘導(dǎo)CD4+CD25+FOXP3+調(diào)節(jié)性T細(xì)胞(Regulatory T cell,Treg)生成的能力;Q-PCR檢測(cè)DCs中多種細(xì)胞因子的表達(dá);流式細(xì)胞術(shù)(FCM)檢測(cè)DC表面分子CD80/CD86、MHCII、CD40的表達(dá)情況。結(jié)果1.重組抗原Eg10和m MDH不能刺激DCs表面樹突的生成,DCs呈現(xiàn)不成熟狀態(tài);2.重組抗原Eg10和m MDH刺激DCs后,IDO的表達(dá)增強(qiáng),上清中犬尿氨酸的含量也升高,且1-MT能特異性降低IDO的表達(dá)水平和酶活性;3.與1-MT未處理的m MDH組相比,1-MT處理的m MDH組中DCs刺激CD4+T細(xì)胞增殖的能力升高,誘導(dǎo)CD4+CD25+FOXP3+Tregs生成的能力降低,差異有統(tǒng)計(jì)學(xué)意義;而Eg10組對(duì)1-MT的反應(yīng)不明顯;4.重組抗原Eg10和m MDH刺激DCs后,TNF-α、IL-6、IL-10 m RNA水平的表達(dá)呈不同程度的升高,1-MT處理的抗原組TNF-αm RNA表達(dá)升高,IL-6、IL-10m RNA表達(dá)降低;5.重組抗原Eg10和m MDH刺激DCs后,細(xì)胞表面分子的表達(dá)水平較低,1-MT處理并不能提高表面分子的表達(dá)水平。結(jié)論1.細(xì)粒棘球蚴重組抗原Eg10和m MDH體外刺激小鼠骨髓來(lái)源的DCs,不能使其發(fā)育成熟,DCs刺激CD4+T細(xì)胞增殖的能力較弱;2.細(xì)粒棘球蚴重組抗原m MDH通過(guò)刺激DCs表達(dá)高水平的IDO,引起免疫耐受。3.細(xì)粒棘球蚴重組抗原Eg10不是通過(guò)刺激DCs表達(dá)IDO引起免疫耐受的,可能存在其他機(jī)制。
[Abstract]:Objective to investigate the mechanism of immune tolerance induced by recombinant antigens Eg10 and m MDH of echinococcus granulosus in mice.Method 1.Dendritic cells derived from mouse bone marrow were cultured in vitro. DCS were stimulated with Eg10 and m MDH in vitro. The morphological changes of DCs were observed by scanning electron microscope. The expression of Indoleamine 23-dioxygenase (IDO) in DCs was observed by immunofluorescence.DCs was treated with 1-Methyl-tryptophane 1-MT-1, a IDO inhibitor, for 3 h to inhibit the expression and enzyme activity of IDO.Then the recombinant antigen Eg10 and m MDH were used to stimulate DCS in vitro, and the ability of DCs to stimulate the proliferation of CD4 T cells and to induce the production of CD4 CD25 FOXP3 regulatory T cell Treg-T cells was detected by mixed lymphocyte reaction. Q-PCR was used to detect the expression of various cytokines in DCs.Flow cytometry (FCM) was used to detect the expression of CD80 / CD86 and MHCII- 鈪，

本文編號(hào)：1737844