【作者】 黄勇；

【导师】 张学光；

【作者基本信息】 苏州大学 ， 免疫学， 2006， 硕士

【摘要】 树突状细胞(DC)是体内已知功能最强的专职性抗原提呈细胞(APC)，能刺激初始型T淋巴细胞活化，从而启动机体特异性免疫应答。DC在诱导特异性抗肿瘤免疫应答中发挥重要的作用，DC疫苗业已用于某些肿瘤的临床免疫治疗。IL-2是最早被发现的淋巴细胞因子之一，不仅在T淋巴细胞的克隆性增殖和活化过程中发挥关键作用，而且可以促进NK细胞和B淋巴细胞的增殖分化。重组IL-2已被临床作为治疗性药物应用。近年来，小鼠DC分泌IL-2的生物学效应已有研究，但IL-2在人外周血单核细胞来源DC(Mo-DC)分化成熟和功能介导中的作用尚未见报导。 本文分析CD40信号和TNF-α对体外诱导Mo-DC表达CD25(IL-2Ra)的调节作用，继而着重研究CD40信号激发的Mo-DC能否表达功能性高亲合力IL-2受体(IL-2R)，在此基础上观察外源性IL-2对Mo-DC分化发育和功能的影响，以此探讨IL-2／IL-2R在调节Mo-DC分化、发育和成熟过程中介导的作用，为Mo-DC疫苗的研制及其在免疫治疗中的应用提供新的思路。 第一部分 CD40信号对单核细胞来源DC表达IL-2R／IL-2的诱导作用及其相关生物学效应 目的：比较抗人CD40激发型单抗和TNF-α对Mo-DC体外诱导表达CD25(IL-2Rα)的作用，以及Mo-DC表面高亲合力IL-2R的表达，并进一步探讨外源性IL-2在Mo-DC分化成熟和功能介导中的生物学效应。 方法：采用联合GM-CSF和IL-4体外诱导Mo-DC的培养方法，分别加入CD40单抗和TNF-α诱导Mo-DC成熟；免疫荧光标记分析Mo-DC表型；Westernblotting检测Mo-DC NF-κB核转位；共聚焦荧光显微镜和免疫共沉淀检测高亲和力IL-2受体的表达；细胞免疫组化检测IL-2的分泌；ELISA法和胞浆细胞因子测定IFN-γ的分泌水平；~3H-TdR掺入法分析Mo-DC对自体T细胞的促增殖效应。 结果：(1) CD40信号能有效的诱导成熟Mo-DC上调表达CD25(IL-2Rα)，而TNF-α更多还原

【Abstract】 Dendritic cells (DC) are professional APC that are specialized for the initiation and regulation of T cell immunity, especially the potent antitumor immunity. Now, DC vaccines has been applied to treat tumor in many clinic trials of immunotherapy. IL-2 is a cytokine which is able to sustain the growth of T, B and natural killer (NK) cells and, during the late phases of antigen specific T-cell response, it contributes to the maintenance of T-cell homeostasis by promoting activation-induced cell death (AICD) of effector T lymphocytes. Recombinant IL-2 demonstrated a value regent in clinic therapy. The effect of IL-2 on human monocyte-derived DC still need to be illustrated.In this study, we investigated the distinct expression profile of CD25 on CD40 and TNF-a activated Mo-DC, and determinate whether CD40 activated DC do express functional IL-2 receptor. Basing on above, we demonstrated the role of IL-2/IL-2R in differentiation, maturation and function of Mo-DC, which could provide value data in preparing DC vaccine.Part I : CD40 signal induced Mo-DC expression of IL-2R/IL-2 and the related biological effectsObjects: compare the roles of CD40mAb or TNF-a induced CD25 (IL-2Ra) expressionon Mo-DC in vitro, study on highly avidity IL-2R expressed on Mo-DC surface, roles ofIL-2 on Mo-DC generation and functions.Methods: using GM-CSF, IL-4 to induce Mo-DC as usual, CD40 or TNF-a was addedinto mature DC; FACS analyzed Mo-DC phenotypes; westernblotting detected Mo-DCNF-kB; confocal microscope and immunoprecipitation detected highly avidity IL-2Rexpression; cytohistochemistry detected IL-2; ELISA and FACS measured IFN-γ inplasma; ~3H-TdR measured T cell proliferation.Results: (1) CD40 could up-regulate the expression of CD25 (IL-2Ra) on Mo-DC, after更多还原