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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71207完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 莊雅惠(Ya-Hui Chuang) | |
| dc.contributor.author | Hung-Wen Chen | en |
| dc.contributor.author | 陳虹汶 | zh_TW |
| dc.date.accessioned | 2021-06-17T04:58:38Z | - |
| dc.date.available | 2028-07-26 | |
| dc.date.copyright | 2018-08-01 | |
| dc.date.issued | 2018 | |
| dc.date.submitted | 2018-07-26 | |
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Liu, X., et al., A protective role of IL-30 via STAT and ERK signaling pathways in macrophage-mediated inflammation. Biochem Biophys Res Commun, 2013. 435(2): p. 306-12. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71207 | - |
| dc.description.abstract | 原發性膽汁性膽管炎(Primary biliary cholangitis, PBC)是一個肝臟慢性自體免疫疾病，在我們先前的研究中發現將攜帶IFN-γ基因的Adeno-associated virus (AAV)給予小鼠的同時誘發小鼠產生PBC，在疾病早期與對照組小鼠相比發炎程度升高很多，但在疾病晚期可能因為Interlukin-30 (IL-30)的上升使IFN-γ造成的疾病嚴重程度趨緩，因此推論IL-30在PBC小鼠中可能具有免疫抑制功能。IL-30，是IL-27的次單元體，又稱為IL-27p28，其他研究指出IL-30可以減緩肝臟發炎所造成的損傷與纖維化。因此在本研究中我們將AAV-mIL-30打入2-OA-OVA誘發的PBC小鼠模式，探討IL-30在原發性膽汁性膽管炎中是否具有免疫抑制功能。首先，我們使用已被廣泛研究的conA誘發之急性肝損傷小鼠模型確認AAV-mIL-30在體內的免疫抑制能力，發現給予AAV-mIL-30的急性肝損傷小鼠其血清中的IFN-γ與IL-12與對照組小鼠相比明顯降低，顯示AAV-mIL-30在體內確實有免疫抑制的能力。在PBC中，我們使用2-OA-OVA誘發小鼠產生PBC後三週再給予AAV-mIL-30，兩週後分析肝臟浸潤免疫細胞的數目與功能，發現IL-30可以降低CD4+ T細胞的活化與IFN-γ的分泌，且抗原呈獻細胞的功能被抑制，而CD4+ Foxp3+ 調節性T細胞數目增加，因此我們認為IL-30是透過抑制抗原呈獻細胞與增加調節性T細胞來達到抑制T細胞的效果。另外，我們也發現在給予AAV-mIL-30小鼠肝臟中類鐸受體(Toll-like receptor, TLR)的表現量下降。而在11週PBC小鼠中我們發現給予AAV-mIL-30小鼠相較於對照組小鼠，肝臟中第一型纖維蛋白的表現量下降，但是肝臟門脈區淋巴細胞浸潤的情形並沒有被減緩。由以上實驗結果顯示IL-30在PBC中可以抑制T細胞的活化與分泌IFN-γ的能力，且可以增加調節性T細胞。 | zh_TW |
| dc.description.abstract | Primary biliary cholangitis (PBC) is a chronic liver autoimmune disease. Our previous study found that PBC mice administered with adeno-associated virus-expressing IFN-γ (AAV-IFN-γ) showed a severe disease performance in early phase but subsequently led to downregulation of chronic inflammation with an increase of interleukin-30 (IL-30). IL-30, also called IL-27p28, has been shown to attenuate liver injury and fibrosis. In this study, we investigated whether IL-30 had an immunosuppressive function in PBC by administering mouse IL-30 expressing AAV (AAV-mIL-30) to 2-OA-OVA immunized PBC mice. At first, we defined the immunosuppressive function of AAV-mIL-30 in vivo by a well-known conA-induced hepatitis mouse model. The results showed that serum levels of IFN-γ and IL-12 were decreased in AAV-mIL-30 receiving conA induced hepatitis mice, indicating that AAV-mIL-30 had an immunosuppressive function in vivo. In PBC, the expression of CD25 and IFN-γ secretion in CD4+ T cells were decreased in mice administered with AAV-mIL-30 three weeks post 2-OA-OVA immunization. The suppressive function of IL-30 might be due to the increase the frequency of CD4+Foxp3+ regulatory T cells and inhibition of the function of antigen presenting cells. In addition, we also found the expression of toll-like receptors (TLRs) in liver was decreased in AAV-mIL-30 receiving PBC mice. Moreover, the type I collagen production was decreased while the lymphocytes infiltration in portal area was not changed in AAV-mIL-30 injected mice 11 weeks post 2-OA-OVA immunization. These results suggested that IL-30 could suppress the function of T cells and increase the number of Tregs in PBC. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T04:58:38Z (GMT). No. of bitstreams: 1 ntu-107-R05424004-1.pdf: 2213665 bytes, checksum: ef1edfa7cc3c83b1206726d582cd29dd (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | 致謝 i
摘要 ii Abstract iii Abbreviation v Content vii Content of Figures x Content of Tables xii Chapter 1. Introduction 1 1.1 Primary biliary cholangitis (PBC) 2 1.2 Adaptive immune response in PBC 3 1.2.1 Humoral immune response 3 1.2.2 Cellular immune response 4 1.3 Innate immunity in PBC 5 1.4 Xenobiotic-induced PBC murine model 6 1.5 Immune response in xenobiotic-induced PBC mice 7 1.6 IL-12 family cytokines 8 1.7 Biological function of Interleukin-30 (IL-30) 8 1.8 IL-30 and autoimmune diseases 9 1.9 IL-30 and other diseases 9 1.10 Adeno-associated virus (AAV) 10 1.11 AAV-DJ Helper Free Bicistronic Expression System (GFP) 11 1.12 Concanavalin A (ConA) induced hepatitis murine model 11 Specific aims 13 Chapter 2. Materials and Methods 14 2.1 Mice 15 2.2 AAV-mIL-30 packaging 15 2.3 AAV purification 16 2.4 AAV concentration 17 2.5 AAV titration 17 2.6 AAV injection 18 2.7 Serum sampling 18 2.8 Enzyme-Linked Immunosorbent Assay (ELISA) 19 2.9 Liver RNA extraction and cDNA synthesis 19 2.10 Quantitative real-time PCR (qRT-PCR) 20 2.11 Liver perfusion and pathological section 20 2.12 Masson’s trichrome (TRI) stain 21 2.13 Isolation of liver leukocytes 21 2.14 ConA induced hepatitis murine model 22 2.15 2-OA-OVA/α-GalCer induced PBC murine model 22 2.16 Serum anti-PDC-E2 IgG and IgM level 23 2.17 Serum cytokines detection 24 2.18 Cell surface staining 24 2.19 Intracellular cytokine detection 25 2.20 Intracellular staining (ICS) 25 2.21 Functional assay of antigen presenting cells (APCs) 26 2.22 Intracellular staining for Foxp3 26 2.23 Flow cytometry analysis 27 2.24 Statistical analysis 28 Chapter 3. Results 29 3.1 No significant inflammation response in naïve mice injected with AAV-mIL-30.30 3.2 AAV-mIL-30 had immunosuppressive function in conA-induced hepatitis murine model. 31 3.3 IL-30 in PBC murine model. 31 3.3.1 IL-30 decreased the activation of T cells. 31 3.3.2 IL-30 decreased the IFN-γ secretion of CD4+ T cells. 32 3.3.3 IL-30 decreased the proportion of PMNs. 32 3.3.4 IL-30 did not affect the antibody production of B cells. 33 3.4 The possible mechanism of IL-30 in PBC. 33 3.4.1 IL-30 did not increase the expression of IL-10 in liver. 33 3.4.2 IL-30 did not affect the DCs population and antigen presenting function. 34 3.4.3 IL-30 decreased the ability of cytokine production in liver leukocytes activated with TLR agonists. 34 3.4.4 IL-30 decreased the mRNA expression of TLR4 and TLR7 in liver. 35 3.5 The therapeutic effect of IL-30 in late stage PBC. 35 3.5.1 IL-30 did not attenuate the liver injury. 36 3.5.2 IL-30 slightly decreased the expression of collagen production in liver. 36 3.5.3 IL-30 increased the percentage and the number of CD4+ Foxp3+ regulatory T cells. 36 3.5.4 IL-30 seems to increase the serum expression of various cytokines and chemokines after 11 weeks post immunization. 37 3.6 Conclusion 38 Chapter 4. Discussion 39 Figures 46 Tables 68 Reference 71 Appendix 79 | |
| dc.language.iso | en | |
| dc.subject | 自體免疫疾病 | zh_TW |
| dc.subject | 原發性膽汁性膽管炎 | zh_TW |
| dc.subject | CD4+ T細胞 | zh_TW |
| dc.subject | AAV | zh_TW |
| dc.subject | 免疫療法 | zh_TW |
| dc.subject | IL-30 | zh_TW |
| dc.subject | CD4+ T cells. | en |
| dc.subject | autoimmune disease | en |
| dc.subject | Interleukin-30 | en |
| dc.subject | immune therapy | en |
| dc.subject | AAV | en |
| dc.subject | Primary biliary cholangitis | en |
| dc.title | 探討IL-30於原發性膽汁性膽管炎之免疫調控作用 | zh_TW |
| dc.title | Investigating the Immunoregulatory Effects of Interleukin-30 on Primary Biliary Cholangitis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 106-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蘇剛毅,張永祺,沈家瑞 | |
| dc.subject.keyword | 原發性膽汁性膽管炎,自體免疫疾病,IL-30,免疫療法,AAV,CD4+ T細胞, | zh_TW |
| dc.subject.keyword | Primary biliary cholangitis,autoimmune disease,Interleukin-30,immune therapy,AAV,CD4+ T cells., | en |
| dc.relation.page | 83 | |
| dc.identifier.doi | 10.6342/NTU201801995 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2018-07-26 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 醫學檢驗暨生物技術學研究所 | zh_TW |
| 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-107-1.pdf 未授權公開取用 | 2.16 MB | Adobe PDF |
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