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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 陳俊任(Chun-Jen Chen) | |
dc.contributor.author | Yao-Hsuan Yu | en |
dc.contributor.author | 游耀萱 | zh_TW |
dc.date.accessioned | 2021-06-15T07:00:17Z | - |
dc.date.available | 2016-02-20 | |
dc.date.copyright | 2011-02-20 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-01-24 | |
dc.identifier.citation | 1.許瑞祥: 靈芝概論. In. 台北市: 萬年出版社; 1993.
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Int J Immunopharmacol 1991, 13(8):1109-1115. 36.Kino K, Mizumoto K, Sone T, Yamaji T, Watanabe J, Yamashita A, Yamaoka K, Shimizu K, Ko K, Tsunoo H: An immunomodulating protein, Ling Zhi-8 (LZ-8) prevents insulitis in non-obese diabetic mice. Diabetologia 1990, 33(12):713-718. 37.Haak-Frendscho M, Kino K, Sone T, Jardieu P: Ling Zhi-8: a novel T cell mitogen induces cytokine production and upregulation of ICAM-1 expression. Cell Immunol 1993, 150(1):101-113. 38.Lin YL, Liang YC, Tseng YS, Huang HY, Chou SY, Hseu RS, Huang CT, Chiang BL: An immunomodulatory protein, Ling Zhi-8, induced activation and maturation of human monocyte-derived dendritic cells by the NF-kappaB and MAPK pathways. 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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/48520 | - |
dc.description.abstract | 由不同真菌純化出與 LZ-8序列相近的免疫調節蛋白質,通稱為 FIPs (fungal immunomodulatory proteins),其中 GMI 是近年來成功選殖自小孢子靈芝 (Ganoderma microsporum) 的免疫調節蛋白質。本實驗室發現 GMI 刺激小鼠巨噬細胞株 RAW264.7 分泌 TNF-α,增加一氧化氮生成,並增進吞噬作用 (phagocytosis),因此本篇研究以小鼠感染李斯特菌 (Listeria monocytogenes) 的模式,探討 GMI 對抗微生物感染之能力,結果顯示先給予小鼠 GMI 可以有效的降低脾臟和肝臟的菌數量,抑制細菌的感染。此外在動物實驗中,利用 GMI 免疫調節蛋白質作為免疫佐劑,以卵清蛋白 (OVA) 為抗原對小鼠進行免疫,發現 GMI 可增加抗原專一性的免疫球蛋白 IgG1 和 IgG2a及 Th 1 反應細胞激素之 IFN-γ 生成。以 OVA 刺激免疫過後之小鼠脾臟細胞,能刺激專一性的細胞增生,增加CD4和CD8 T細胞表面活化標誌CD44表現量,並引發專一性的CD8 T細胞毒殺作用:提升 granzyme B在CD8 T細胞中表現量及殺死表現有 OVA257-264 peptide (MHC class Ι epitops) 的目標細胞。進一步以 OVA 與GMI進行免疫,可使 MO5 腫瘤 (表現 OVA 之 B16 黑色素瘤) 之生長受到抑制。 雖然 GMI 在小鼠體內具有免疫佐劑活性,但以 GMI 刺激小鼠骨髓細胞衍生之樹突細胞 (BMDC) 之結果顯示, GMI 不能刺激增加樹突細胞的表面分子 CD40、CD80、CD86、MHC II 的表現量,也無法讓樹突細胞分泌 TNF-α 。由本研究結果顯示,選殖自小孢子靈芝之GMI蛋白,藉由巨噬細胞的活化,可增加小鼠抵抗微生物感染的能力。 此外 GMI 可作為免疫佐劑,促使免疫反應趨向 Th1反應,並活化CD4和CD8 T細胞,引發專一性的細胞毒殺作用。 | zh_TW |
dc.description.abstract | The immunomodulatory protein GMI was recently cloned from Ganoderma microsporum. We have previously found that GMI could stimulate murine monocyte macrophage RAW264.7 cells to produce TNF-α and NO and enhanced phagocytosis. In this study, we first evaluated the potential antimicrobial activity of GMI against Listeria monocytogenes infection in mice, and we found that GMI pretreatment significantly reduced the bacterial loads in liver and spleen after L. monocytogenes infection. We also investigated whether GMI could function as an adjuvant in vivo. Mice immunized with the OVA antigen and GMI generated OVA-specific IgG1 and IgG2a, and after OVA restimulation, the splenocytes underwent proliferation, produced IFN-γ, and had enhanced expression of CD44 on both CD4 and CD8 T lymphocytes. GMI and OVA immunization also induced an OVA-specific CTL response, as demonstrated by higher levels of granzyme B expression in CTLs and specific lysis of OVA257-264 peptide-pulsed target cells. Furthermore, mice immunized with OVA and GMI were also protected from MO5 (OVA-transfected B16 melanoma) tumor challenge. Although GMI stimulates the activation of macrophages, interestingly we could not detect a stimulatory effect of GMI on bone marrow-derived dendritic cells (BMDCs). Taken together, our results demonstrated that GMI could stimulate an innate immune response which protected mice from L. monocytogenes infection as well as function as an adjuvant that promoted the activation of Th1 and CTL responses. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T07:00:17Z (GMT). No. of bitstreams: 1 ntu-100-R97b47112-1.pdf: 1952140 bytes, checksum: 4b5f7dd2134df79b54d736c719d4d3e8 (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 論文口試委員審定書 i
致謝 ii 中文摘要 iii Abstract iv Abbreviation vi 目錄 vii 圖表目錄 x 第一章 總論 1 1. 靈芝之簡介 2 1.1 靈芝的成份及藥理活性 2 1.1.1 多醣體 ( polysaccharides) 3 1.1.2 三萜類 (triterpenoids) 3 1.1.3 真菌類免疫調節蛋白質 3 2. 免疫反應 (immunity) 4 2.1 先天性免疫 (innate immunity) 4 2.1.1 吞噬細胞之活化 4 2.1.2 發炎反應 (inflammatory response) 4 2.2 後天性免疫 (adaptive immunity) 5 2.2.1 細胞免疫反應 (cell-mediated immunity) 5 2.2.2 體液免疫反應 (humoral immunity) 6 2.3 抗原呈現細胞 (antigen-present cells, APC) 7 2.4 李斯細菌 (Listeria monocytogenes) 感染引發之免疫反應 8 2.5 佐劑 (adjuvant) 9 3. 靈芝免疫調節蛋白質 9 3.1 Ling Zhi-8 ( LZ-8 ) 9 3.2 Ganoderma microsporum immunomodulatory protein 10 第二章 研究動機與目的 12 1. GMI在動物體內對微生物感染之影響 13 2. GMI 作為免疫佐劑之活性 13 3. 以 MO5 腫瘤模式評估 GMI作為佐劑引發之細胞毒殺作用 13 4. 探討 GMI是否能刺激樹突細胞之成熟 14 第三章 材料與方法 15 1. GMI 材料介紹 16 2. 實驗用小鼠 16 3. 細胞培養 16 4. GMI 在動物體內對微生物感染之影響 16 4.1 培養李斯特菌 (Listeria monocytogenes ) 16 4.2 評估GMI抗微生物感染之小鼠模式 17 4.3 脾臟和肝臟菌數之分析 17 4.4 統計與繪圖軟體之分析 18 5. 以GMI 作為免疫佐劑之活性 18 5.1 GMI作為免疫佐劑之小鼠模式 18 5.2 去除OVA抗原內毒素 19 5.3 收集血清樣品進行具OVA專一性抗體測定 19 5.4 小鼠初代脾臟細胞培養與刺激 20 5.5 細胞激素濃度之測定 20 5.6 脾臟細胞增生試驗 21 5.7 以流式細胞儀分析 T 細胞表面 CD44 之表現量 21 5.8 以流式細胞儀分析CD8 T細胞之granzyme B表現量 21 5.9 細胞毒殺作用測試 (in vitro killing) 22 5.10 統計與繪圖軟體之分析 23 6. 以 MO5 腫瘤模式評估 GMI 作為佐劑引發之細胞毒殺作用 23 6.1 MO5 (B16-OVA) 腫瘤細胞 23 6.2 小鼠免疫與腫瘤模式 23 7. 探討 GMI 是否能刺激樹突細胞之成熟 24 7.1 BMDC之培養與刺激 24 7.2 細胞激素濃度之測定 24 7.3 BMDC成熟標誌之分析 25 7.4 統計與繪圖軟體之分析 25 第四章 實驗結果 26 1 GMI 在動物體內對微生物感染之影響 27 1.1 李斯特菌感染小鼠之致死劑量測試 27 1.2 給予 GMI對小鼠感染李斯特菌之影響 27 2. 以GMI 作為免疫佐劑之活性 27 2.1 以GMI作為佐劑可增加 OVA-specific IgG1 及IgG2a之生成 27 2.2 以GMI 作為佐劑可增加細胞激素 IFN-γ 之生成 28 2.3 以 GMI 作為佐劑可刺激具抗原專一性之淋巴球活化與增生 28 2.4 以GMI作為佐劑進行免疫後可增加 T 細胞表面CD44之表現量 28 2.5 以GMI作為佐劑可活化CD8 T 細胞之細胞毒殺作用 29 3. 以 MO5 腫瘤模式評估 GMI 作為佐劑引發之細胞毒殺作用 29 3.1 以 OVA 併 GMI 對小鼠進行免疫,可有效的抑制 MO5 腫瘤的生 長 29 4. 探討 GMI 是否能刺激樹突細胞之成熟 30 第五章 結論與討論 31 圖表 34 參考文獻 52 圖表目錄 圖一、已發表真菌類免疫調節蛋白胺基酸序列比對 35 圖二、 FIP-gts 之胺基酸序列與二級結構 36 圖三、免疫調節蛋白 LZ-8 (A) 與 GMI (B) 之三級結構 37 圖四、以SDS-PAGE分析pichia pastoris所生產之重組蛋白質GMI 38 圖五、給予 GMI 之小鼠感染李斯特菌 48小時後,對肝臟與脾臟菌數 量之影響 39 圖六、以 GMI 作為佐劑對血清中 OVA-specific IgG1 及IgG2a 生成之影響 40 圖七、以 GMI作為佐劑對細胞激素 IFN-γ 及 IL-17A 生成之影響41 圖八、以 GMI 作為佐劑刺激脾臟細胞之情形 42 圖九、以 GMI 作為佐劑增加 CD4 T 細胞上表面 CD44 表現量 43 圖十、以 GMI 作為佐劑增加 CD8 T 細胞上表面 CD44 表現量 44 圖十一、以 GMI 作為佐劑增加 CD8 T 細胞中 granzyme B 表現之情 形 45 圖十二、以 GMI 作為佐劑對細胞毒殺作用之影響 46 圖十三、以 GMI 作為佐劑對 MO 5抗腫瘤之效果 47 圖十四、以 GMI 作為佐劑各組帶有腫瘤的小鼠百分比 48 圖十五、以 GMI 不能誘導小鼠骨髓細胞分化之樹突細胞表面成熟標 誌表現 49 表一、以 GMI 刺激 BMDC 表面成熟 marker 之情形之影響 50 圖十六、以 GMI 刺激小鼠骨髓細胞分化之樹突細胞對細胞激素 TNF-α 生成之影響 51 | |
dc.language.iso | zh-TW | |
dc.title | Ganoderma microsporum 免疫調節蛋白GMI 在動物體之免疫佐劑與抗微生物感染活性之研究 | zh_TW |
dc.title | Antimicrobial and adjuvant functions of the immunomodulatory protein GMI from Ganoderma microsporum in animals | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 許秉寧,徐立中 | |
dc.subject.keyword | 免疫調節蛋白質,佐劑, | zh_TW |
dc.subject.keyword | fungal immunomodulatory protein,Ganoderma microsporum,antimicrobia l,Listeria monocytogenes,adjuvant, | en |
dc.relation.page | 56 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-01-24 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
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