【摘要】：Toll樣受體(TLR)是固有性免疫應(yīng)答中的重要成分，不但在感染性免疫中發(fā)揮重要作用，，而且參與了腫瘤的免疫應(yīng)答和免疫逃逸過程，TLR家族中受到廣泛關(guān)注的是TLR4分子。樹突狀細(xì)胞(DC)作為機(jī)體免疫應(yīng)答的始動者，在對多種疾病應(yīng)答中，尤其是在對腫瘤的免疫應(yīng)答中發(fā)揮了不可替代的作用。已有研究表明多種因素可以影響單核細(xì)胞及樹突狀細(xì)胞上TLR4的表達(dá)，但共刺激分子CD40和4-1BB對TLR4表達(dá)的影響尚未見報(bào)道。本研究旨在通過激活DC或單核細(xì)胞上CD40或4-1BB分子后，觀察TLR4分子表達(dá)的變化，以期為腫瘤免疫治療提供新的方法與思路。 1．4-1BB信號對小鼠樹突狀細(xì)胞TLR4表達(dá)的調(diào)節(jié) 目的：探討激發(fā)型小鼠4-1BB(CD137)單克隆抗體2A對小鼠骨髓來源未成熟DC(imDC)及樹突狀細(xì)胞株DC2.4表面TLR4表達(dá)的調(diào)節(jié)。方法：在imDC中加入不同劑量2A(聯(lián)合兔抗大鼠IgG-Fc段抗體，CL)、LPS、2A(+CL)與LPS聯(lián)合后作用6h，流式細(xì)胞術(shù)(FCM)檢測TLR4表達(dá)；或在以上處理因素作用后不同時相(0h、3h、6h、12h、24h)，以及先用LPS刺激imDC 6h再加入2A(+CL)作用6h后，以FCM檢測imDC表面TLR4表達(dá)。結(jié)果：LPS可下調(diào)imDC上TLR4的表達(dá)，作用可維持24h。單獨(dú)使用2A無明顯效應(yīng)，但與CL聯(lián)合后2A可使imDC上調(diào)TLR4分子的表達(dá)，作用可維持12h。2A(+CL)與LPS聯(lián)合作用仍可使imDC上調(diào)表達(dá)TLR4分子。先用LPS預(yù)處理imDC6h再加入2A作用6h，2A仍可拮抗LPS的下調(diào)作用。在樹突狀細(xì)胞株DC2.4細(xì)胞表面也得到相似的結(jié)果。結(jié)論：4-1BB信號可上調(diào)小鼠骨髓來源未成熟DC和樹突狀細(xì)胞株DC2.4表面TLR4的表達(dá)，上調(diào)效應(yīng)具有時間和劑量依賴性，并可拮抗LPS介導(dǎo)的TLR4下調(diào)效應(yīng)。 2．CD40信號對人單核細(xì)胞和樹突狀細(xì)胞TLR4表達(dá)的調(diào)節(jié) 目的：探討人激發(fā)型CD40單克隆抗體5C11對人單核細(xì)胞和樹突狀細(xì)胞TLR4
[Abstract]:Toll like receptor (TLR) is an important component of innate immune response, which not only plays an important role in infectious immunity, but also plays an important role in tumor immune response and immune escape. As the initiator of immune response, dendritic cell (DC) plays an irreplaceable role in the response to various diseases, especially in the immune response to tumor. Several factors have been shown to affect the expression of TLR4 in monocytes and dendritic cells, but the effects of costimulatory molecules CD40 and 4-1BB on the expression of TLR4 have not been reported. The purpose of this study was to observe the expression of CD40 or 4-1BB on DC or monocytes and to provide new methods and ideas for tumor immunotherapy. Regulation of TLR4 expression in murine dendritic cells by 1.4-1BB signal objective: to investigate the effect of mouse 4-1BB (CD137) monoclonal antibody 2A on mouse bone marrow Regulation of TLR4 expression on DC2.4 surface of mature DC (imDC) and dendritic cell lines. Methods: different dosages of 2A (combined with rabbit anti-rat IgG-Fc fragment antibody CL) were added to imDC for 6 h after treatment with LPS and TLR4 expression was detected by flow cytometry (FCM). The expression of TLR4 on imDC surface was detected by FCM at different time (0 h ~ 3 h ~ 6 h ~ 12 h ~ (24 h) after the treatment of above factors, and imDC was stimulated by LPS for 6 h and then treated with 2A (CL) for 6 h. The expression of TLR4 on the surface of imDC was detected by FCM. Results the expression of TLR4 on imDC could be down-regulated by lipopolysaccharide (LPS) for 24 h. Using 2A alone had no obvious effect, but 2A combined with CL could up-regulate the expression of TLR4 molecule in imDC, and maintain the combination of 12h.2A (CL) and LPS, which could make imDC upregulate the expression of TLR4 molecule. Pretreatment of imDC6h with LPS and addition of 2A at 6 h could still antagonize the down-regulation of LPS. Similar results were obtained on the surface of dendritic cell line DC2.4. Conclusion the expression of TLR4 on the surface of murine bone marrow-derived immature DC and dendritic cell line DC2.4 can be up-regulated by the signal of: 4-1BB in a time-and dose-dependent manner, and can antagonize the down-regulation of TLR4 mediated by LPS. Regulation of TLR4 expression in human monocytes and dendritic cells by 2.CD40 signal objective: to investigate the effect of human stimulated CD40 monoclonal antibody 5C11 on human mononuclear fine cells Cell and dendritic cell TLR4
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 富寧;Toll樣受體與病原相關(guān)分子模式[J];上海免疫學(xué)雜志;2001年04期

2 郝軼群;Toll樣受體與樹突狀細(xì)胞[J];上海免疫學(xué)雜志;2003年03期

本文編號：2259278