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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 許輔 | |
dc.contributor.author | Ju-Chun Hsu | en |
dc.contributor.author | 許如君 | zh_TW |
dc.date.accessioned | 2021-06-13T06:52:58Z | - |
dc.date.available | 2007-08-01 | |
dc.date.copyright | 2005-08-01 | |
dc.date.issued | 2005 | |
dc.date.submitted | 2005-07-27 | |
dc.identifier.citation | 參考文獻
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/35441 | - |
dc.description.abstract | 摘要
本研究探討台灣金線連免疫調節蛋白(immunomodulatory protein from A. formosanus Hayata, IPAF)之生化特性及免疫調節活性,並進一步瞭解IPAF在免疫細胞中可能之訊息傳導路徑。台灣金線連經30%硫酸銨沈澱可得分子量為14 kDa之IPAF蛋白,名為台灣金線連免疫調節蛋白,等電點約為5.4。IPAF能活化RAW 264.7巨噬細胞產生一氧化氮(nitric oxide, NO),並能刺激TNF-α及IL-1β的產生。IPAF亦可活化淋巴細胞,提升細胞激素IFN-γ的分泌量,以及刺激小鼠脾臟細胞的增生,但不引起IL-4的產生。經由流式細胞儀分析得知小鼠脾臟細胞主要之增生細胞群為B細胞,並於去除T細胞之環境下仍能受IPAF刺激進行增生反應,故認為IPAF屬於胸腺非依賴性抗原(thymus-independent antigen)。在訊息傳導路徑方面,C57BL/10ScN小鼠試驗發現IPAF刺激TLR4-/-腹腔巨噬細胞所產生之TNF-α分泌量降為TLR4+/+腹腔巨噬細胞分泌量之7.35 %,故推測IPAF於巨噬細胞中之訊息傳導路徑與TLR4高度相關;另一方面,IPAF在去除T細胞之環境下仍能使TLR4-/-之B細胞增生,故認為IPAF在B細胞中之訊息傳導路徑與TLR4無明顯相關,應有另一路徑傳遞其增生訊息。綜合實驗結果可知IPAF能活化B細胞,並能經由TLR4活化巨噬細胞,可提升宿主免疫反應,極具研究開發價值。 | zh_TW |
dc.description.abstract | Abstract
IPAF (immunomodulatory protein from Anoectochilus formosanus Hayata) was purified from Anoectochilus formosanus Hayata by 30% saturation of ammonium sulfate precipitation. SDS-PAGE analysis showed that IPAF has a molecular weight of 14 kDa and isoelectric focusing electrophoresis revealed the isoelectric point of IPAF was 5.4. Culturing RAW 264.7 macrophages with IPAF in vitro showed that IPAF could activate the cells and then increase nitric oxide, TNF-α and IL-1β production. Moreover, IPAF also stimulated the proliferation of murine splenocytes and increased their IFN-γ secretion, but did not result in IL-4 production. However, the cell depletion test demonstrated that IPAF increased the B cell proliferation without T cells existing. These results suggested that IPAF was a thymus-independent antigen. Peritoneal cells from C57BL/10ScN mice, which have a null mutation in TLR4 (TLR4-/-), were hyporesponsive in TNF-α secretion to both LPS and IPAF, suggesting that LPS and IPAF may share a same signaling pathway involving TLR4. On the other hand, IPAF could increase TLR4-/- B cells proliferation. Therefore, we propose that IPAF might be not involved in TLR4 signaling pathway and have another signaling pathway in B cell proliferation. Taking together, this study clearly demonstrated that IPAF, which activated B cells and also activated macrophages through TLR4, is an immune stimulus and could be helpful to strength the immunity of its host. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T06:52:58Z (GMT). No. of bitstreams: 1 ntu-94-R91628212-1.pdf: 1430259 bytes, checksum: f5ed2fb80f3b70798c3a957e66e53b44 (MD5) Previous issue date: 2005 | en |
dc.description.tableofcontents | 目錄
中文摘要 1 英文摘要 2 第一章 緒論 1-1 前言 3 1-2 金線連簡介 4 1-2-1 金線連之種類及生長形態 5 1-2-2 金線連之生長與栽培 5 1-2-3 金線連之繁殖方法 6 1-2-4 台灣金線連之生理活性 6 1-2-5 台灣金線連之毒性測試 12 1-2-6 台灣金線連IFAF之生理活性 14 1-3 先天性免疫反應與Toll-like receptor 4介紹 18 1-3-1先天性免疫反應(Innate immune response) 19 1-3-2 Toll-like receptors簡介 19 1-3-3 Toll-like receptor 4之相關分子研究及其訊息傳導路徑 22 1-3-3-1 Toll-like receptor 4之相關分子研究 22 1-3-3-2 MyD88 dependent signaling pathway介紹 24 1-3-3-3 MyD88 independent signaling pathway介紹 26 1-4 B細胞及T細胞之活化因子 28 1-4-1 B細胞之活化 28 1-4-1-1體液性免疫反應(humoral immune response) 28 1-4-1-2 B細胞的活化 29 1-4-2 T細胞之活化與分化 31 1-4-2-1 T細胞之活化與增生 31 1-4-2-2 CD4 T細胞的分化 34 1-4-2-3 CD8 T細胞的分化 34 1-5 研究動機與構想 35 第二章 材料與方法 2-1 台灣金線連蛋白之純化及濃度測定 37 2-1-1台灣金線連蛋白IPAF之純化 38 2-1-2 IPAF蛋白質濃度測定 38 2-2 台灣金線連蛋白IPAF之特性分析 39 2-2-1 膠體電泳分析(SDS-polyacrylamide slab gel electrophoresis) 40 2-2-2西方轉漬分析(Western blot) 42 2-2-3等電點分析 44 2-3台灣金線連蛋白IPAF對免疫細胞活性及增生之影響 45 2-3-1 IPAF對巨噬細胞RAW 264.7之影響 48 2-3-1-1 IPAF之PMB及LAL試驗 48 2-3-1-2一氧化氮(NO)試驗 50 2-3-1-3 TNF-α試驗 52 2-3-1-4 IL-1β分析 53 2-3-2 IPAF對小鼠脾臟細胞之影響 54 2-3-2-1細胞激素IFN-γ之測定 55 2-3-2-2細胞激素IL-4之測定 56 2-3-2-3 BrdU攝取分析 57 2-3-2-4利用流式細胞儀分析IPAF對B細胞及T細胞 增生之影響 58 2-3-2-5利用microbead純化法分析IPAF對非B(non-B)、 非T(non-T)細胞之影響 60 2-4台灣金線連蛋白IPAF在免疫細胞中之訊息傳導路徑 62 2-4-1 IPAF在巨噬細胞中可能之訊息傳導路徑與鍵結關係 64 2-4-1-1 C57BL/10ScN小鼠試驗 65 2-4-1-2 IPAF與RAW 264.7巨噬細胞之鍵結關係 66 2-4-2 IPAF對B細胞之活化與TLR4之關係 67 第三章 結果 3-1 台灣金線連蛋白之純化及特性分析 69 3-1-1台灣金線連蛋白之純化與全光譜分析(UV/VIS absorption spectrum analysis) 69 3-1-2膠體電泳分析(SDS-polyacrylamide slab gel electrophoresis) 70 3-1-3西方轉漬分析(Western blot) 71 3-1-4 IPAF之蛋白質濃度測定 72 3-1-5等電點分析 72 3-2台灣金線連蛋白IPAF對免疫細胞活性及增生之影響 73 3-2-1 IPAF對巨噬細胞RAW 264.7之影響 73 3-2-1-1 IPAF之PMB及LAL試驗 73 3-2-1-2一氧化氮(NO)及TNF-α試驗 74 3-2-1-3 IL-1β分析 76 3-2-2 IPAF對小鼠脾臟細胞之影響 76 3-2-2-1細胞激素IFN-γ之測定 76 3-2-2-2細胞激素IL-4之測定 77 3-2-2-3 BrdU攝取分析 78 3-2-2-4利用流式細胞儀分析IPAF對B細胞及 T細胞增生之影響 79 3-2-2-5利用microbead純化法及BrdU攝入分析法探討IPAF對T細胞、B細胞、非B(non-B)、非T(non-T)細胞之影響 81 3-2-2-6利用microbead純化法及流式細胞儀分析探討IPAF 對非B(non-B)、非T(non-T)細胞之影響 83 3-3台灣金線連蛋白IPAF在免疫細胞中可能之訊息傳導路徑 85 3-3-1 IPAF在巨噬細胞中可能之訊息傳導路徑與鍵結關係 85 3-3-1-1 C57BL/10ScN小鼠試驗 85 3-3-1-2 IPAF與RAW 264.7巨噬細胞之鍵結關係 87 3-3-2 IPAF對B細胞之活化與TLR4之關係 88 第四章 討論 91 參考文獻 96 圖 107 附錄 131 | |
dc.language.iso | zh-TW | |
dc.title | 台灣金線連免疫調節蛋白IPAF活化免疫細胞之機制 | zh_TW |
dc.title | Activating mechanism of Immunomodulatory Protein from Anoectochius formosanus (IPAF) on immune cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 93-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 柯俊良,沈立言,繆希椿 | |
dc.subject.keyword | 台灣金線連,活化,免疫細胞, | zh_TW |
dc.subject.keyword | Anoectochilus formosanus,activation,immune cells, | en |
dc.relation.page | 138 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2005-07-28 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 園藝學研究所 | zh_TW |
顯示於系所單位: | 園藝暨景觀學系 |
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