本文選題：Th17 + 細(xì)胞　； 參考：《南京醫(yī)科大學(xué)》2012年博士論文

【摘要】：本論文分為兩個(gè)部分。第一部分主要探討Th17細(xì)胞及白細(xì)胞介素-17（IL-17）與小鼠異基因造血干細(xì)胞移植后移植物抗宿主�。℅VHD）之間的關(guān)系；第二部分則探索了一種基于基因工程技術(shù)制備和分選Th17細(xì)胞的新方法。 Th17細(xì)胞是近年提出的一個(gè)輔助性T細(xì)胞新亞群，主要通過分泌IL-17等細(xì)胞因子發(fā)揮功能，與抗感染免疫和自身免疫病發(fā)生密切相關(guān)。然而，Th17細(xì)胞與GVHD的關(guān)系至今仍有很大爭(zhēng)論。在第一部分研究中，我們首先通過輸注不同的脾細(xì)胞量建立了重度和輕-中度GVHD小鼠模型，發(fā)現(xiàn)外周血Th17細(xì)胞的比率和IL-17水平與GVHD嚴(yán)重程度成負(fù)相關(guān)關(guān)系。我們進(jìn)一步細(xì)化了allo-HSCT后CD4+T細(xì)胞亞群失衡與不同靶臟器損傷的相關(guān)性，發(fā)現(xiàn)Th1細(xì)胞與肝臟損傷相關(guān)，Th17細(xì)胞與肺損傷相關(guān)，Th1/Th17細(xì)胞比率與小腸損傷相關(guān)。為了進(jìn)一步驗(yàn)證Th17細(xì)胞與GVHD相關(guān)肺損傷的關(guān)系，我們使用了肺特異性高表達(dá)IL-17的轉(zhuǎn)基因小鼠作為受鼠建立GVHD模型，，不僅證實(shí)IL-17是GVHD相關(guān)肺損傷的促進(jìn)因素，而且發(fā)現(xiàn)IL-17系通過促進(jìn)肺組織表達(dá)趨化因子增加炎癥細(xì)胞招募和促進(jìn)Th1細(xì)胞分化加重GVHD相關(guān)肺損傷，初步闡明了其作用機(jī)制。 Th17細(xì)胞由于含量很低且無(wú)膜表面特異性標(biāo)志或標(biāo)記組合，又由于其發(fā)育調(diào)控的復(fù)雜性導(dǎo)致體外誘導(dǎo)產(chǎn)率不高而難以制備和純化。在第二部分研究中，我們嘗試將過表達(dá)Th17細(xì)胞發(fā)育的主要正調(diào)控因子RORγt與TGF-β+IL-6細(xì)胞因子體外處理聯(lián)合起來(lái)，使naive CD4+T細(xì)胞獲得了更高的Th17細(xì)胞得率。此外，我們?cè)谳d體上負(fù)載了IL-17啟動(dòng)子驅(qū)動(dòng)的截短型人神經(jīng)生長(zhǎng)因子受體（ΔNGFR）報(bào)告基因，通過分選ΔNGFR+細(xì)胞純化了制備的基因工程Th17細(xì)胞。表型和功能鑒定表明，過表達(dá)RORγt聯(lián)合TGF-β+IL-6細(xì)胞因子體外處理制備的Th17細(xì)胞較單純過表達(dá)RORγt制備的Th17細(xì)胞更接近天然的Th17細(xì)胞，可為Th17細(xì)胞功能的研究提供有效的工具。
[Abstract]:This paper is divided into two parts. In the first part, the relationship between Th17 cells, interleukin-17 (IL-17) and graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation was studied. In the second part, a new method for preparing and sorting Th17 cells based on genetic engineering is explored. Th17 cells are a new helper T cell subgroup proposed in recent years. Th17 cells play a role by secreting cytokines such as IL-17. It is closely related to the occurrence of anti-infection immunity and autoimmune disease. However, the relationship between Th17 cells and GVHD is still controversial. In the first part of the study, we first established the severe and mild-moderate GVHD model by injecting different amount of spleen cells. We found that the ratio of Th17 cells and IL-17 level in peripheral blood were negatively correlated with the severity of GVHD. We further refined the relationship between the imbalance of CD4 T cell subsets after allo-HSCT and the injury of different target organs. It was found that the ratio of Th1 cells and liver injury related to Th17 cells and lung injury correlated with the ratio of Th1 / Th17 cells and intestinal injury. In order to further verify the relationship between Th17 cells and GVHD-associated lung injury, we used transgenic mice with lung specific high expression of IL-17 as a GVHD model to establish GVHD model, which not only confirmed that IL-17 was the promoting factor of GVHD-associated lung injury. It was also found that IL-17 increased the recruitment of inflammatory cells and enhanced the differentiation of Th1 cells by promoting the expression of chemokines in lung tissue. Th17 cells are difficult to be prepared and purified due to their low content and lack of membrane surface specific markers or marker combinations, and because of the complexity of their developmental regulation, resulting in a low induced yield in vitro. In the second part of the study, we try to combine ROR 緯 t, the main positive regulatory factor of over-expression Th17 cell development, with TGF- 尾 IL-6 cytokines in vitro, so that naive CD4 T cells can obtain higher Th17 cell yield. In addition, we loaded truncated human nerve growth factor receptor (螖 NGFR) reporter gene driven by IL-17 promoter on the vector and purified the engineered Th17 cells by sorting 螖 NGFR cells. Phenotypic and functional identification showed that Th17 cells prepared by overexpression of ROR 緯 t combined with TGF- 尾 IL-6 cytokines were closer to natural Th17 cells than those prepared by over-expressing ROR 緯 t in vitro, which could provide an effective tool for the study of Th17 cell function.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R392.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Inhibiting the expression of CD28 costimulatory molecule on human lymphocytes by special siRNA[J];Chinese Medical Journal;2005年06期

本文編號(hào)：2105688