請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/10747
標題: | 探討過敏性紫斑症之Th1與Th17 cells之角色及建立小鼠動物模式 The Role of Th1/Th17 Cells and Establishment of Mouse Model in Henoch-Schonlein Purpura |
作者: | Hsiao-Yu Jen 任曉玉 |
指導教授: | 莊雅惠(Ya-Hui Chuang) |
關鍵字: | 過敏性紫斑症,Th1 cells,Th17 cells,human β2GPI, HSP,Th1 cells,Th17 cells,human β2GPI, |
出版年 : | 2010 |
學位: | 碩士 |
摘要: | 過敏性紫斑症 (Henoch-Schonlein Purpura; HSP) 是孩童常見的全身性微血管發炎之自體免疫疾病。在體液性免疫方面,急性期病人的血清IgA升高,TNF-α、IL-6、IL-8、TGF-β與VEGF亦升高,且出現多種IgA 自體抗體 (IgA autoantibodies),但在細胞免疫方面的研究則相當缺乏。此外,目前亦無動物模式可用於研究其致病機轉。在本研究探討T helper (Th)1與Th17 cells之角色及嘗試破除免疫耐受性以建立自體免疫疾病之小鼠模式。
第一部分 探討過敏性紫斑症中Th1與Th17 cells之角色 Th1與Th17 cells之角色在許多自體免疫疾病已被證實。在此我們探討Th1與Th17 cells在過敏性紫斑症之角色。首先我們測病人血清中Th1與Th17相關細胞激素IL-17、IL-21、IL-23與IFN-γ之表現量。結果顯示,相較於正常對照組,急性期過敏性紫斑症病人血清中IL-17表現量顯著增加,但IFN-γ並無差異。此外,急性過敏性紫斑症病人之週邊血單核球IL-17-secreting T cells (Th17 cells)與上清液中IL-17含量均比緩解期時增加;然而IFN-γ-secreting T cells (Th1 cells)與上清液中IFN-γ則無明顯差異。由此推論,過敏性紫斑症可能是Th17主導的自體免疫疾病,而與Th1免疫反應較無相關。 第二部分 建立過敏性紫斑症小鼠動物模式 β2GPI被認為是過敏性紫斑症的自體抗原之一,在我們以注射human β2GPI (hβ2GPI)以建立小鼠疾病模式。將human β2GPI混合於佐劑以腹腔注射致敏小鼠。結果顯示,hβ2GPI混合於CFA/IFA中致敏小鼠,小鼠可產生高量之anti-mouse β2GPI (mβ2GPI)IgG,但anti-mβ2GPI IgA並無增加,病理檢驗結果亦未見血管發炎。再以hβ2GPI與先前研究證實可以提高血清IgA的Deoxynivalenol (DON)同時致敏小鼠仍然只能引起anti-mβ2GPI IgG,而無anti-mβ2GPI IgA。改以Alum當佐劑仍然無法引起小鼠anti-mβ2GPI IgA反應。綜合言之,注射human β2GPI可以打破免疫耐受性使小鼠產生anti-mouse β2GPI IgG,但仍需其他方法以引發IgA反應。 由本研究的結果得知,過敏性紫斑症可能是Th17主導的自體免疫疾病,而與Th1免疫反應較無相關。另外,注射human β2GPI可以打破小鼠免疫耐受性,若可建立此過敏性紫斑症之小鼠模式,則有利於後續更深入之研究。 Henoch-Schonlein Purpura (HSP) is an autoimmune vasculitis that occurs mainly in childhood. At the acute stage of HSP, serum levels of IgA, TNF-α, IL-6, IL-8, TGF-β, and VEGF were increased. In addition, a number of studies report that several IgA autoantibodies are associated with HSP. However, there were no reports regarding cellular immunity in HSP and no HSP animal models so far to explore the mechanism of HSP. Herein, the aims of this study are to investigate the role of Th1 and Th17 cells in HSP and try to establish a mouse model of HSP. Part I : The role of Th1 and Th17 cells in Henoch-Schonlein Purpura Recently, various studies demonstrated that Th1 and Th17 are important in the pathogenesis of several autoimmune diseases, such as multiple sclerosis, rheumatoid arthritis, and Crohn’s disease. The role of Th1 and Th17 cells has not been investigated in HSP. Therefore, the aim of this study is to examine the role of Th1 and Th17 cells in HSP. Our results demonstrated that serum levels of IL-17 were significantly elevated in patients with acute stage of HSP compared with healthy controls, whereas serum levels of IFN-γ remained unchanged. Furthermore, the percentage of Th17 cells and the cytokine levels of IL-17 were increased at the acute stage compared with those at the convalescent stage by using intracellular staining for Th1 and Th17 cell detection and ELISA for culture supernatant detection. However, the percentage of Th1 cells and IFN-γ expression were not elevated at the acute stage of HSP. These results suggested that IL-17 rather than IFN-γ play a crucial role in the pathogenesis of HSP. Part II : Establishment of mouse model of Henoch-Schonlein Purpura β2-glycoprotein I (β2GPI) are supposed to be an autoantigen of HSP because high levels of IgA anti-β2GPI antibodies were observed in patients with HSP. The specific aim of this study is to establish a mouse model of HSP. We firstly immunized naïve mice with human β2GPI (hβ2GPI) protein emulsified in adjuvant. Our results showed that hβ2GPI-immunized mice had elevated serum levels of anti-mβ2GPI IgG but not anti-mβ2GPI IgA. In addition, the biopsies have not seen inflammation in the wall of capillaries. To induce a high serum level of IgA, we used hβ2GPI and deoxynivalenol (DON), a mycotoxin can elevate serum IgA and mesangial IgA deposition in mice, or hβ2GPI emulsified in aluminum to immunize mice. However, the serum levels of β2GPI-specific IgA were not elevated in both strategies. In conclusion, our results demonstrated that IL-17 may play an important role in the pathogenesis of HSP. Moreover, establishment of the mouse model of HSP can provide the further study to determine the mechanism of HSP. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/10747 |
全文授權: | 同意授權(全球公開) |
顯示於系所單位: | 醫學檢驗暨生物技術學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-99-1.pdf | 2.69 MB | Adobe PDF | 檢視/開啟 |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。