探討Notch ligand Delta4對樹突狀細胞的影響與應用於氣喘之研究

• Main Author: 林鉅倫
• Other Authors: 醫學科學研究所
• Language: Chinese; English
• Published: 2010
• Subjects: 樹突狀細胞; Dent; Weaver; Osborne; Barton; Johansen; McIntyre; Ramos; Rousseau; Fang; Buckley; Jagged; Ramsay; Bertrand; Mellor; Wharton; Chiang; Janus; Lira; Yvon; Corry; Kanda; Kuipers; Brockton; Dent La; Crassostrea gigas; Mite
• Online Access: http://libir.tmu.edu.tw/handle/987654321/36417

謝誌 4 中文摘要 6 英文摘要 8 縮寫對照表 10 第一章 序論 12 （一）Notch的介紹 13 （二）Notch及Notch ligand的組成 14 （三）Notch的訊息傳遞 15 （四）Notch對樹突狀細胞分化的影響 15 （五）Notch對T細胞分化的影響 17 （六）氣喘疾病的發生 20 （七）細胞激素在氣喘中扮演之角色 21 （八）樹突狀細胞於氣喘反應之角色 22 （九）氣喘的治療 23 第二章 研究動機與目的 25 第三章 探討重組蛋白rmDelta4對樹突狀細胞活化之影響以及是否可藉由此修飾後之樹突狀細胞調控CD4+ T細胞之分化 28 （一）實驗目的與原理 29 （二）實驗材料與方法 30 （三）研究結果 37 第四章 探討經重組蛋白rmDelta4刺激後之樹突狀細胞在氣喘動物模式中的預防效果 41 （一）實驗目的與原理 42 （二）實驗材料與方法 43 （三）研究結果 48 第五章 討論 51 第六章 結論與未來方向 56 圖 59 參考文獻 69 Wharton KA, Johansen KM, Xu T, Artavanis-Tsakonas S. 1985. Nucleotide sequence from the neurogenic locus notch implies a gene product that shares homology with proteins containing EGF-like repeats. Cell 43: 567-81 2. Tsukumo S, Yasutomo K. 2004. Notch governing mature T cell differentiation. J Immunol 173: 7109-13 3. Cheng P, Nefedova Y, Corzo CA, Gabrilovich DI. 2007. Regulation of dendritic-cell differentiation by bone marrow stroma via different Notch ligands. Blood 109: 507-15 4. Fiuza UM, Arias AM. 2007. Cell and molecular biology of Notch. 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Respir Res 7: 72 Notch訊息可維持細胞增生、分化及凋亡間的平衡，影響許多器官的發育和功能，在生物體中扮演著很重要的角色。樹突細胞為抗原呈獻細胞的一種，在抗原刺激後會促進T細胞的活化及naïve CD4+ T細胞的分化。一般認為Notch ligands中的Delta4跟抗原呈獻細胞衍生的Th1細胞分化有關，但Nocth訊息中複雜的交互作用，以及在樹突細胞的發育及分化、還有T細胞的活化與Th細胞的分化仍尚未完全明白。在本實驗中，我們假設Delta4對於調控樹突狀細胞的活化及功能扮有很重要的角色。體外實驗中可以觀察到，若在樹突狀細胞發育的過程中給予Delta4刺激會造成樹突狀細胞表面分子MHC class Ⅱ的表現量下降但CD80及CD86的表現量則有上升的情形。另外，我們也觀察到這種經Delta4修飾過後的樹突狀細胞分泌細胞激素IL-10、IL-12、IL-23及IL-27與前發炎激素IL-1β、IL-6及TNF-α皆有下降的情形，因此我們推測這一種樹突狀細胞為一種半成熟的樹突狀細胞。另外，將這群修飾後的樹突狀細胞與T細胞共培養，我們觀察到T細胞的增生情形會被抑制，同時細胞激素IL-5及IFN-γ的分泌量也會減少。此外，我們進一步探討在氣喘動物模式下給予經Delta4作用後的樹突狀細胞是否能預防氣喘疾病的發生。由實驗結果發現，經由Delta4作用的樹突狀細胞能降低OVA專一性的IgE抗體的產生，同時減少氣管呼吸道阻力、肺部發炎細胞浸潤及細胞激素的產生。綜合以上所述，我們推測這一群經由Delta4刺激後的樹突狀細胞確實能有效的降低氣喘的發炎反應，因此未來或許可利用此修飾的樹突狀細胞來進行氣喘的治療。 The Notch pathway participates in cell proliferation, differentiation, and apoptosis processes that affects the development and function in various organs. Dendritic cells (DCs), as professional antigen presenting cells (APCs), induce T cell activation and promote T cells differentiation by antigen stimulation. It is known that Delta4 on APCs is associated with the stimulation of Th1-type response. However, the regulation of activation and differentiation in DCs and T cells by Delta4 remains unclear. In this study, we firstly investigated whether Delta4 played an immune-modulatory role in the activation and function of DCs in vitro. The DCs were treated with Delta4 during cells development and show low levels of MHC class Ⅱ but high level of CD80 and CD86 expression in Delta4-treated DCs. These modified DCs might be semi-mature form of DCs because they expressed low level of cytokines such as IL-12,IL-23,IL-27,IL-1β,IL-6,IL-10 and TNF-α. In addition, these Delta4-treated DCs not only inhibited the proliferation of T cells, but also reduced the production of IL-5 and IFN-γ of activated T cells. Furthermore, these modified DCs was applied to examine the preventive effects in established asthmatic animal model. In here, we showed that these modified DCs efficently moderated the characteristics of asthma, including expression of OVA-specific IgE antibody, airway hyper-responsiveness, eosinophilic airway inflammation, and cytokine production. Taken together, these results suggest that Delta4-modulated DCs are effective in suppressing asthmatic airway inflammation and are a potential therapeutic tool for asthma.