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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80161完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王汎熒(Fun-In Wang) | |
| dc.contributor.author | Jia-Ling Yang | en |
| dc.contributor.author | 楊佳玲 | zh_TW |
| dc.date.accessioned | 2022-11-23T09:29:24Z | - |
| dc.date.available | 2021-08-06 | |
| dc.date.available | 2022-11-23T09:29:24Z | - |
| dc.date.copyright | 2021-08-06 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-07-11 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80161 | - |
| dc.description.abstract | "藍舌病(Bluetongue, BT)因造成反芻獸出血性疾病，對畜牧產業衝擊嚴重。世界動物衛生組織(OIE)將BT列為須通報疾病，許多國家重視BT的防治與疫苗開發。藍舌病病毒(bluetongue virus, BTV)屬於呼腸孤病毒科(Reoviridae)的Orbivirus屬，通過昆蟲媒介傳播。迄今全世界已經確認了29種BTV血清型。西元2003年，台灣也分離出兩種BTV毒株，分別為BTV2 / KM / 2003和BTV12 / PT / 2003。由於BTV毒株間差異可能引發不同強度的臨床症狀，因此本論文旨在比較該兩種病毒株與其宿主免疫細胞的交互作用，觀察各病毒株間的差異。在第二章，我們將BTV2分別接種牛隻週邊血液單核細胞(peripheral blood mononuclear cell, PBMC)、單核球來源巨噬細胞(monocyte-derived macrophage, MDM)及巨噬細胞重建自體淋巴球細胞(MDM-Lymphocyte)後，觀察BTV的複製及細胞素的變化。此三類細胞群分別代表了BTV進入宿主體內，會遭遇的不同細胞族群。BTV2的複製曲線在PBMC、MDM與MDM-Lymphocyte中，三者相似且無顯著差異。在PBMC與MDM-Lymphocyte中，IL-4 mRNA的表現早於先天免疫和細胞介導免疫的細胞素。培養細胞上清液中IL-4蛋白的穩定存在與IgE抗體的出現，顯示第I型過敏反應參與了致病機轉。然而在MDM(無淋巴細胞)中，IL-1β，TNF-α和IL-12p40的mRNA表現均未受抑制。此結果支持了淋巴球與IL-4可能對先天免疫與細胞介導免疫的抑制作用。值得注意的是，即使在相同物種中，各BTV血清型之間的致病機轉也不同，因此在第三章，我們比較了BTV2和BTV12在PBMC中免疫路徑的差異。BTV2優先刺激IL-4的早期表現(T輔助2路徑)，並可能反饋抑制先天免疫。相反地，BTV12優先刺激先天免疫，以誘導TNF-α和IL-1β。BTV非結構蛋白3抗體(抗BTV-NS3)螢光信號顯示PBMC和MDM中的單核球是BTV複製的首選細胞。生物資訊學分析顯示誘導IL-4的能力可能位於VP2蛋白三維結構的尖端區域(tip region)。其中VP2尖端區上，預測肽段的數量與BTV感染中報導過的過敏反應呈正相關。BTV2-VP2的合成肽可於12-48小時，誘導IL-4及IgE表現，而BTV12的合成肽則無，與生物資訊學預測一致。合成肽刺激牛PBMC似乎是BTV誘導過敏反應的良好模型，此方法應可加速病毒蛋白致病機轉和發展疫苗的應用研究。" | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-23T09:29:24Z (GMT). No. of bitstreams: 1 U0001-2806202121031500.pdf: 4462907 bytes, checksum: 5caea5e7cec900bb1bbae0419640a84a (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "Certificate i Acknowledgement ii 中文摘要 iii Abstract iv Content vi Lists of Figures ix Lists of Tables x Chapter One. General Introduction 1 1.1 Background 1 1.2 The virion structure and viral replication 4 1.3 Viral-host immunity 8 1.3.1 Innate immunity 9 1.3.2 Humoral immune response (Th2 pathway) 11 1.3.3 cell-mediated immunity (Th1 pathway) 12 1.4 Bioinformatics 16 1.4.1 MHC class II binding protein prediction 16 1.4.2 IL-4 inducing peptide prediction by IL-4pred 17 1.4.3 The three-dimensional (3D) protein structure modeling of VP2 18 1.5 The aim of this study 18 Chapter Two. Type I Hypersensitivity Is Induced in Cattle PBMC during Bluetongue Virus Taiwan Isolate Infection 19 2.1 Introduction 19 2.2 Materials and methods 21 2.2.1 Experimental design 21 2.2.2 Preparation of virus, bovine PBMC, MDM and MDM-Lymphocyte 22 2.2.3 Quantitation of intracellular BTV loads represented by core protein VP7 genome RNA and load of progeny virus in cell associated culture supernatant 24 2.2.4 Sample RNA extraction and relative quantitation of cytokine expression 25 2.2.5 Quantitation of Cytokine and IgE protein levels by Enzyme-linked immunosorbent assay (ELISA) 28 2.2.6 Transferrable anti-viral activity by quantitating BTV loads in recipients 28 2.2.7 Statistical analysis 28 2.3 Results 29 2.3.1 Yields of PBMC and MDM 29 2.3.2 BTV2 viral load increases gradually from 12 to 48 hpi in PBMC, MDM and MDM-Lymphocyte 31 2.3.3 BTV2 infection restrained innate immunity in PBMC and MDM-Lymphocyte but not in MDM 33 2.3.4 BTV2 induces more dominant Th2 cytokine expression than that of the Th1 pathway 35 2.3.5 The significant protein expressions of IL-4 and IgE suggest a bias toward Th2 and the engagement of type I hypersensitivity during BTV2 infection 36 2.3.6 Transferrable anti-viral activity by quantitating BTV loads in recipient infected PBMC from other cows 37 2.4 Discussion 39 2.5 Conclusions 43 Chapter Three. The Tip Region on VP2 Protein of Bluetongue Virus Contains Potential IL-4-Inducing Amino Acid Peptide Segments 44 3.1 Introduction 44 3.2 Materials and Methods 48 3.2.1 Animals and Preparation of Bovine PBMC and Monocyte-Derived Macrophage (MDM) 48 3.2.2 Virus Infection and Quantitation of BTV Viral Loads in PBMC 49 3.2.3 TCID50 Assay to Determine the Titers of Progeny Virus in the PBMC Supernatant 49 3.2.4 Pair-Wise Comparison of the Cytokine mRNA Expression by qRT-PCR 50 3.2.5 IFA of BTV-Infected MDM by Confocal Microscopy 51 3.2.6 Apoptosis Detection Assay 52 3.2.7 Enzyme-Linked Immunosorbent Assay (ELISA) for IL-1β, IFN-α, IL-4 and IgE 52 3.2.8 The Prediction of Potential IL-4-Inducing Peptides and Three-Dimensional (3D) Protein Structure Modeling on VP2 of BTVs 53 3.2.9 Synthesis of BTV Potential IL-4 Inducing Peptides 53 3.2.10 PBMC Stimulation Assay for Synthetic IL-4-Inducing Peptides 54 3.2.11 Statistical Analysis 55 3.3 Results 55 3.3.1 Experimental Design 55 3.3.2 The Viral Loads and Titers of BTV2/BTV12 in Infected PBMC 56 3.3.3 Infection Rates of BTV2 and BTV12 in MDM Detected by IFA 58 3.3.4 BTVs induce apoptosis, cell aggregation, with or without syncytium in MDM 59 3.3.5 Expression of mRNA in BTV-Infected PBMC 62 3.3.6 The Cytokine Production Induced by BTV2 and BTV12 63 3.3.7 Prediction of Potential IL-4-Inducing Peptides on the VP2 of Various BTV Strains 65 3.3.8 Predicted IL-4-Inducing Peptide Segments in the VP2 Tip Region of BTV2 and BTV12 70 3.3.9 Induction of IL-4 Production in PBMC Stimulated with Synthetic Peptide 72 3.4 Discussion 73 3.5 Conclusions 79 Chapter Four. General Discussion 80 Chapter Five. Conclusion and Perspectives 84 References 85 Appendices 105 List of Published Papers 126 國立臺灣大學獸醫專業學院獸醫學研究所博士學位口試紀錄 127" | |
| dc.language.iso | en | |
| dc.subject | 生物資訊學 | zh_TW |
| dc.subject | 藍舌病 | zh_TW |
| dc.subject | 週邊血液單核細胞 | zh_TW |
| dc.subject | 巨噬細胞 | zh_TW |
| dc.subject | 細胞素 | zh_TW |
| dc.subject | 致病機轉 | zh_TW |
| dc.subject | monocyte-derived macrophage | en |
| dc.subject | bluetongue virus | en |
| dc.subject | bioinformatics | en |
| dc.subject | pathogenesis | en |
| dc.subject | cytokines | en |
| dc.subject | peripheral blood mononuclear cell | en |
| dc.title | 臺灣藍舌病第二血清型病毒株BTV2/KM/2003引起牛隻免疫細胞第一型過敏反應之探討 | zh_TW |
| dc.title | Type I Hypersensitivity in Bovine Immune Cells Induced by Bluetongue Virus Taiwan Isolate BTV2/KM/2003 | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.advisor-orcid | 王汎熒(0000-0001-9597-5182) | |
| dc.contributor.coadvisor | 王家琪(Chia-Chi Wang) | |
| dc.contributor.coadvisor-orcid | 王家琪(0000-0002-1272-1513) | |
| dc.contributor.oralexamcommittee | 李龍湖(Hsin-Tsai Liu),宣詩玲(Chih-Yang Tseng),張銘煌,張家宜 | |
| dc.subject.keyword | 藍舌病,週邊血液單核細胞,巨噬細胞,細胞素,致病機轉,生物資訊學, | zh_TW |
| dc.subject.keyword | bluetongue virus,peripheral blood mononuclear cell,monocyte-derived macrophage,cytokines,pathogenesis,bioinformatics, | en |
| dc.relation.page | 134 | |
| dc.identifier.doi | 10.6342/NTU202101179 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2021-07-12 | |
| dc.contributor.author-college | 獸醫專業學院 | zh_TW |
| dc.contributor.author-dept | 獸醫學研究所 | zh_TW |
| 顯示於系所單位： | 獸醫學系 | |
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