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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳秀男 | |
| dc.contributor.author | Po-Wen Chang | en |
| dc.contributor.author | 鄭博文 | zh_TW |
| dc.date.accessioned | 2021-06-15T16:38:51Z | - |
| dc.date.available | 2020-08-20 | |
| dc.date.copyright | 2015-08-20 | |
| dc.date.issued | 2015 | |
| dc.date.submitted | 2015-08-11 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/53006 | - |
| dc.description.abstract | 虹彩病毒科裡共有五屬，其中細胞腫大病毒屬 (Megalocystovirus)對養殖漁業造成危害最為嚴重。本實驗是採用DNA疫苗來做為防治虹彩病毒的方法。先前以虹彩病毒錐體膜蛋白基因 (OSGIV-8L)作為抗原基因之DNA疫苗，另外導入一段點帶石斑魚細胞白介素1-β (Interleukin-1β, IL-1β)基因作為免疫佐劑，但並未執行攻毒試驗，因此本實驗測試IL-1β基因做為免疫佐劑的功效進而評估提升疫苗免疫效果。將本實驗DNA疫苗轉染 (Transfect)進石斑魚鰭細胞結果發現，細胞確實會表現出抗原蛋白。同樣的結果也在活體實驗中發現，接種DNA疫苗後石斑魚肌肉、脾臟在組織免疫化學染色 (Immunohistochemical, IHC）結果中發現皆有抗原被偵測到，其中又以含免疫佐劑IL-1β之DNA疫苗表現最多。而對於石斑魚免疫力增強部分，在實驗淋巴細胞吞噬能力、超氧陰離子產生立、溶菌酶活性中搭配IL-1β使用的DNA疫苗表現皆較控制組、不含免疫佐劑DNA組來的好。魚血清中專一性抗體的量亦是含免疫佐劑IL-1β之DNA疫苗組別最多。且IL-1β基因能誘導TNF-α、CD8α、IL-8之基因表現，進而在攻毒試驗中展現了良好的存活力 (90 %)。本實驗結果顯示，搭配免疫佐劑IL-1β基因之DNA疫苗能提升疫苗免疫效果。且IL-1β基因可能藉由提升石斑魚非特異性免疫、刺激免疫相關基因，進一步使DNA疫苗能更迅速低誘發特異性免疫反應，並使石斑魚對點帶石斑虹彩病毒產生抵抗力。這些結果顯示，細胞白介素1β作為DNA疫苗之基因免疫佐劑 (genetic adjuvants)的潛力。 | zh_TW |
| dc.description.abstract | Megalocytivirus is one of iridoviridae family and emerged in recent years as a troublesome pathogen to a wide range of marine and freshwater fish. Vaccination is one of the useful methods against viral disease. In previous study, we used a bicistronic vector encoding OSGIV-8L and grouper interleukin-1β (IL-1β) as a DNA vaccine. The expression of grouper IL-1β was applied to increase the efficiency of DNA vaccine. But, we didn't evaluate how much efficiency increased by IL-1β. So we aimed to determine the efficiency and function of IL-1β in DNA vaccine in this study. In the premiere results, the grouper fin cells were transfected with the DNA vaccine and the antigen protein were detected through western blot. Moreover, in the in vivo experiments, after vaccinating via intramuscular injection, OSGIV-8L's mRNA and protein were detected in spleen and muscle. Among the experimental conditions, the DNA vaccine containing IL-1β gene express more OSGIV-8L's mRNA and protein than the DNA vaccine without IL-1β gene. Furthermore, the immunological analysis showed that pIRES-8L/IL-1β was able to (Ⅰ) stimulate the respiratory burst of lymphocytes at 21 day post-vaccination, (Ⅱ) stimulate the phagocytic capacity of lymphocytes at 3day and 21day post-vaccination, (III) stimulate the lysozyme activity at 7day,14day and 21day post-vaccination, (IV) upregulate the expression of immune related genes, and (V) induce the production of serum antibodies. And when challenged groupers with 104 copies of OSGIV, the groupers vaccinated with pIRES-8L/IL-1β showed higher survive rate compared to control groupers and the groupers vaccinated with DNA vaccine without IL-1β gene. In summary, the DNA vaccine with IL-1β gene may enhance the grouper nonspecific immunity to induce specific immune response more rapidly, and develop resistance to virus. These results reveal that the potential application of fish IL-1β in fish vaccination. | en |
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| dc.description.tableofcontents | 中文摘要 I
Abstract II 目錄 IV 圖目錄 VI 第一章 文獻整理 1 第一節 世界及台灣水產養殖現況 1 第二節 點帶石斑的習性與養殖現況 2 第三節 虹彩病毒簡介 2 第四節 海水魚類虹彩病毒感染症狀 3 第五節 魚類免疫概述 4 第六節 魚類疫苗概述 6 第七節 OSGIV-8L抗原分析 8 第八節 細胞白介素-1β 8 第九節 研究目的 8 第二章 材料與方法 10 第一節 DNA疫苗製備 10 1.1 限制酶切割 (Restriction enzyme digestion) 10 1.2 膠體純化 (Gel Elution) 10 1.3 電腦軟體比對與分析 10 1.4 大腸桿菌 (Escherichia coli , E.coli)勝任細胞製作 11 1.5 質體轉型作用 (Transformation） 11 1.6 質體純化 (Plasmid Extraction) 11 1.7 DNA疫苗確認 12 第二節DNA疫苗之細胞實驗 12 2.1實驗細胞 12 2.2細胞轉染 (Transfect)及採樣 12 2.3 DNA疫苗在於石斑魚鰭細胞 (In Vitro)之表現 13 第三節DNA疫苗之動物實驗 17 3.1 DNA疫苗在點帶石斑魚體內 (In Vivo)之表現-基因層次 17 3.2 DNA疫苗在點帶石斑魚體內 (In vivo)之表現-蛋白質層次 19 3.3 DNA疫苗對點帶石斑魚免疫能力之影響 20 3.4 DNA疫苗對點帶石斑血清中專一性抗體之影響 23 3.5 DNA疫苗對點帶石斑免疫相關基因影響 24 3.6 DNA疫苗保護力測試 25 第四節 統計分析 26 第三章 結果 27 第一節 DNA疫苗製備 27 1.1 OSGIV-8L 核酸序列比對 27 1.2 細胞白介素-1β核酸序列比對 27 1.3 DNA疫苗確認 27 第二節DNA疫苗之細胞實驗 28 2.1 DNA疫苗在石斑魚鰭細胞 （In Vitro）之表現 28 2.2 DNA疫苗對於石斑魚鰭細胞免疫相關基因影響 28 第三節DNA疫苗之動物實驗 29 3.1 DNA疫苗在點帶石斑魚體內 (In Vivo)之表現-基因層次 29 3.2 DNA疫苗在點帶石斑魚體內 (In Vivo)之表現-蛋白質層次 30 3.3 DNA疫苗對點帶石斑免疫能力之影響分析 30 第四章 討論 35 第五章 結論 40 參考文獻 41 圖表附件 53 | |
| dc.language.iso | zh-TW | |
| dc.subject | 點帶石斑魚 | zh_TW |
| dc.subject | 點帶石斑虹彩病毒 | zh_TW |
| dc.subject | DNA疫苗 | zh_TW |
| dc.subject | 免疫佐劑 | zh_TW |
| dc.subject | 細胞白介素1β | zh_TW |
| dc.subject | Orange-spotted grouper | en |
| dc.subject | DNA vaccine | en |
| dc.subject | immune adjuvant | en |
| dc.subject | interleukin-1β | en |
| dc.subject | Orange-spotted grouper iridovirus virus | en |
| dc.title | 細胞白介素-1β基因做為石斑虹彩病毒DNA疫苗佐劑之效果 | zh_TW |
| dc.title | Adjuvant effects of IL-1β on Grouper DNA vaccination against Iridovirus | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 103-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 冉繁華,劉秉忠,廖文亮 | |
| dc.subject.keyword | 點帶石斑魚,點帶石斑虹彩病毒,DNA疫苗,免疫佐劑,細胞白介素1β, | zh_TW |
| dc.subject.keyword | Orange-spotted grouper,Orange-spotted grouper iridovirus virus,DNA vaccine,immune adjuvant,interleukin-1β, | en |
| dc.relation.page | 80 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2015-08-12 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 漁業科學研究所 | zh_TW |
| 顯示於系所單位： | 漁業科學研究所 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-104-1.pdf 未授權公開取用 | 1.31 MB | Adobe PDF |
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。