本文選題：IL-4 + SDF-1α/CXCL12　； 參考：《安徽醫(yī)科大學(xué)》2005年碩士論文

【摘要】：目的:通過探討IL-4和基質(zhì)細(xì)胞源因子(SDF)-1α/CXCL12協(xié)同刺激下,Naive Th細(xì)胞Th1型、Th2型細(xì)胞因子的表達(dá)以及信號(hào)轉(zhuǎn)導(dǎo)通路中關(guān)鍵分子的活化情況,判斷協(xié)同刺激誘導(dǎo)的Th細(xì)胞極化方向,以及其信號(hào)轉(zhuǎn)導(dǎo)通路可能的分子機(jī)制。 方法:使用流式細(xì)胞術(shù)和RT-PCR檢測IL-4和SDF-1α協(xié)同刺激的臍帶血fCB)CD4~+T細(xì)胞中Th1型/Th2型細(xì)胞因子的mRNA和胞內(nèi)蛋白兩個(gè)水平的表達(dá)情況;利用免疫復(fù)合物激酶分析法和免疫印記法,檢測協(xié)同誘導(dǎo)Th2極化過程中Syk激酶和ZAP-70激酶的磷酸化情況以及Cb1家族和NFAT家族活化的情況;使用ZAP-70PNA技術(shù)阻斷ZAP-70的表達(dá),探討ZAP-70激酶在協(xié)同刺激后Th細(xì)胞極化過程中的重要意義;使用EMSA技術(shù)進(jìn)一步探討NFAT轉(zhuǎn)錄因子家族成員的活化情況,明確參與細(xì)胞因子轉(zhuǎn)錄的關(guān)鍵轉(zhuǎn)錄因子。 結(jié)果:流式檢測結(jié)果:用IL-4和SDF-1α/CXCL12刺激8天,CB CD4+T細(xì)胞(Naive Th)轉(zhuǎn)換成Th2細(xì)胞模式(90.3% IL-4陽性);RT-PCR檢測結(jié)果:新鮮分離的CBCD4+T細(xì)胞中的IFN-γ或者IL-4 mRNA大約7.3×10~2或者2.1×10~3拷貝,相應(yīng)的,用IL-4+SDF-1α/CXCL12刺激的細(xì)胞中的IL-4 mRNA大約有1.3×10~4拷貝,IFN-γ mRNA約7.8×10~1拷貝;免疫復(fù)合物激酶分析法和免疫印記檢測結(jié)果:IL-4和SDF-1α/CXCL12協(xié)同刺激的8天內(nèi),ZAP-70激酶和Cbl-b蛋白發(fā)生顯著而穩(wěn)
[Abstract]:Aim: to investigate the expression of Th1 type Th2 cytokines and the activation of key molecules in signal transduction pathway of naive Th cells after co-stimulation of IL-4 and stromal cell source factor SDF- 1 偽 / CXCL12, and to determine the polarization direction of Th cells induced by co-stimulation. And the possible molecular mechanism of its signal transduction pathway. Methods: flow cytometry and RT-PCR were used to detect the expression of mRNA and intracellular protein of Th1 type Th2 cytokines in IL-4 and SDF-1 偽 co-stimulated cord blood fCBC CD4T cells, and immunocomplex kinase assay and imprinting method were used to detect the expression of Th1 type Th2 cytokines. The phosphorylation of Syk kinase and ZAP-70 kinase and the activation of Cb1 family and NFAT family during co-induced Th2 polarization were detected, and the expression of ZAP-70 was blocked by ZAP-70PNA technique to explore the significance of ZAP-70 kinase in the process of co-stimulated Th cell polarization. The activation of family members of NFAT transcription factors was further investigated by EMSA technique, and the key transcription factors involved in cytokine transcription were identified. Results: IL-4 and SDF-1 偽 / CXCL12 were used to stimulate the transformation of CB CD4 T cells into Th2 cell model (90.3% IL-4 positive RT-PCR): IFN- 緯 or IL-4 mRNA was about 7.3 脳 10 ~ (-2) or 2.1 脳 10 ~ (-3) copies of CBCD4 T cells. There were about 1.3 脳 10 ~ (4) copies of IL-4 mRNA in cells stimulated with IL-4 SDF-1 偽 / CXCL12 and about 7.8 脳 10 ~ (-1) copy of IFN- 緯 mRNA. The results of immuno-complex kinase assay and immunological imprinting showed that ZAP-70 kinase and Cbl-b protein were remarkably stable in 8 days after co-stimulation of SDF-1 偽 -IL-4 and SDF-1 偽 / CXCL12.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R392
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本文編號(hào)：1837288