4-1BB和CD40信号对TLR4表达调节的研究

发布时间：2018-10-09 12:07

【摘要】：Toll样受体(TLR)是固有性免疫应答中的重要成分，不但在感染性免疫中发挥重要作用，，而且参与了肿瘤的免疫应答和免疫逃逸过程，TLR家族中受到广泛关注的是TLR4分子。树突状细胞(DC)作为机体免疫应答的始动者，在对多种疾病应答中，尤其是在对肿瘤的免疫应答中发挥了不可替代的作用。已有研究表明多种因素可以影响单核细胞及树突状细胞上TLR4的表达，但共刺激分子CD40和4-1BB对TLR4表达的影响尚未见报道。本研究旨在通过激活DC或单核细胞上CD40或4-1BB分子后，观察TLR4分子表达的变化，以期为肿瘤免疫治疗提供新的方法与思路。 1．4-1BB信号对小鼠树突状细胞TLR4表达的调节目的：探讨激发型小鼠4-1BB(CD137)单克隆抗体2A对小鼠骨髓来源未成熟DC(imDC)及树突状细胞株DC2.4表面TLR4表达的调节。方法：在imDC中加入不同剂量2A(联合兔抗大鼠IgG-Fc段抗体，CL)、LPS、2A(+CL)与LPS联合后作用6h，流式细胞术(FCM)检测TLR4表达；或在以上处理因素作用后不同时相(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检测imDC表面TLR4表达。结果：LPS可下调imDC上TLR4的表达，作用可维持24h。单独使用2A无明显效应，但与CL联合后2A可使imDC上调TLR4分子的表达，作用可维持12h。2A(+CL)与LPS联合作用仍可使imDC上调表达TLR4分子。先用LPS预处理imDC6h再加入2A作用6h，2A仍可拮抗LPS的下调作用。在树突状细胞株DC2.4细胞表面也得到相似的结果。结论：4-1BB信号可上调小鼠骨髓来源未成熟DC和树突状细胞株DC2.4表面TLR4的表达，上调效应具有时间和剂量依赖性，并可拮抗LPS介导的TLR4下调效应。 2．CD40信号对人单核细胞和树突状细胞TLR4表达的调节目的：探讨人激发型CD40单克隆抗体5C11对人单核细胞和树突状细胞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
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

【参考文献】