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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王汎熒 | |
| dc.contributor.author | Chia-Wei Yen | en |
| dc.contributor.author | 顏嘉緯 | zh_TW |
| dc.date.accessioned | 2021-07-11T15:28:49Z | - |
| dc.date.available | 2023-08-21 | |
| dc.date.copyright | 2018-08-21 | |
| dc.date.issued | 2018 | |
| dc.date.submitted | 2018-08-17 | |
| dc.identifier.citation | Akira, S., Uematsu, S., Takeuchi, O., 2006. Pathogen recognition and innate immunity. Cell 124, 783-801.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78914 | - |
| dc.description.abstract | 藍舌病(bluetongue)是藍舌病毒(bluetongue virus; BTV)所引起、由昆蟲媒介傳播的家畜及野生反芻動物疾病，是全球畜牧業都會面臨到的重要經濟威脅之一。台灣地區也曾有牛和山羊的感染紀錄，呈亞臨床狀態，但藍舌病仍是畜牧業的潛在風險。目前對於BTV的相關免疫研究，大多聚焦在曾於西方國家爆發疫情的強毒株。在低致病性毒株研究，例如感受性較綿羊低的反芻宿主之免疫反應，例如臺灣家畜乳牛與山羊等，都是學界目前較少關注的。本研究嘗試以低毒力台灣株(BTV2 / KM / 2003)對牛隻周邊血液單核球(PBMCs)進行攻毒後的免疫反應對時序進程研究(time course study)。以流式細胞儀分析發現，除了時序進程造成的單核細胞(monocyte)分化效應以外，攻毒組別的單核細胞呈現更強的前向角散射光(forward scatter; FSC)與側向角散射光(side scatter; SSC)信號變化，反映了單核細胞因攻毒而產生的外型及內部結構上的變化，並且，於感染後24及48小時約有30 ± 5%活的單核細胞呈現被感染狀態，其細胞膜CD14-FITC+與細胞質內NS3-PE+皆呈陽性。而細胞激素的信使核糖核酸(mRNA)的相對定量分析則顯示，牛隻單核球細胞於攻毒後未能成功表現IL-1β，但在感染初期即能激起顯著的IL-4表現，也代表著動物實體外細胞免疫Th2途徑的啟動，顯示低毒力台灣株BTV2能躲避宿主的先天免疫機制，有利於病毒傳播感染時的初期複製。然而，本實驗尚未能釐清此毒株是否能在台灣牛隻實體內引起顯著IL-4表現與及在動物宿主免疫上扮演的具體角色，尚需進一步研究。 | zh_TW |
| dc.description.abstract | Bluetongue (BT) is an insect-transmitted disease of domestic and wild ruminants caused by bluetongue virus (BTV), and it is one of the economic threats to animal husbandry worldwide. BTV infection in cow and goats has been recorded in Taiwan; although being subclinical, it still poses a potential risk for livestock. Most studies on BTV-induced immune responses focused on the virulent strains of outbreaks in western countries. Few have studied low virulent strains as well as the less susceptible species, like dairy cow and goats in Taiwan. In this study, responses of bovine peripheral blood mononuclear cells (PBMCs) to low virulent Taiwan strains (BTV2/KM/2003) were conducted. Flow cytometry of infected PBMC demonstrated changes in forward scatter (FSC) and side scatter (SSC) in monocyte population, reflecting the morphological alteration in size and internal structure following BTV infection. Approximately 30 ± 5% viable monocytes could be infected by BTV, evidenced by CD14-FITC+/NS3-PE+ dual positivity. Relative quantification of cytokine mRNA showed BTV failed to induced IL-1β yet markedly induce IL-4 expression at early stage of infection, suggestive of the initiation of Th2 pathway of immune cells ex vivo. The ability to avoid innate immunity is beneficial for early replication of BTV in the host. However, the concrete role of IL-4 in cattle in vivo triggered by low virulent Taiwan strain may require further studies. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-11T15:28:49Z (GMT). No. of bitstreams: 1 ntu-107-R04644002-1.pdf: 1236357 bytes, checksum: 121575ef136e87e6a3a7d8c87028baef (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | Certificate i
Acknowledgement ii 中文摘要 iii Abstract iv Content v Lists of figures vii Lists of tables viii 1. Introduction 1 2. Literature review 3 2.1. Background 3 2.2. Host range and clinical manifestation 5 2.3. Pathogenesis 7 2.4. The virion structure and replication 9 2.5. Host immunity 13 2.5.1. Innate immunity 13 2.5.2. Cellular immunity 16 2.5.3. Humoral immunity 17 3. Material and methods 18 3.1. Experimental design 18 3.2. Preparation of virus, bovine PBMC and ad-PBMC 19 3.3. Experimental infection of PBMC 20 3.4. Flow cytometry analysis 20 3.5. Preparation of external BTV VP7 cRNA standard 21 3.5.1. Quantification and serial dilution of cRNA standard 21 3.6. Quantitation of BTV loads in PBMC represented by core protein VP7 genome RNA 22 3.7. Sample RNA Extraction and Relative Quantitation of Cytokines Expression 23 3.8. Enzyme-linked immunosorbent assay (ELISA) for IL-4 24 3.9. Statistical analysis 24 4. Results 24 4.1. Physical characterization of BTV2 challenged peripheral blood mononuclear cells (PBMC) 24 4.2. Sustained yet limited proliferation of BTV2 in bovine PBMC 25 4.3. Restrained innate immune responses against BTV2 25 4.4. Predominant expression of Th2 rather than Th1-related cytokines 26 4.5. Absolute quantification in cytokine yields following BTV2 challenge 26 5. Discussion 27 6. References 30 Figure 1-1. Characterizations of bovine PBMC at various hours postinfection (hpi) infected with BTV at MOI of 1. 49 Figure 1-2. Isotype control for antibodies used in immunofluorescence staining to determine the antigen positive/negative cutoff. 49 Figure 2. Absolute quantification of BTV viral load in PBMC infected with BTV2 at MOI of 1 for 72 hours. 51 Figure 3. BTV2 induces TNFα expression (a) by monocytes in PBMC early in 6 hpi, but fails to induce significant expression of IL-1β (b). This suggests that monocyte functions are suppressed early postinfection. 52 Figure 4. Th2 pathway represented by IL-4 expression is induced significantly by BTV2 (a). Whereas those of Th1 (b, c) are also induced but insignificant compared to that of 0 hpi. 53 Figure 5. Secretion of IL-4 from lymphocytes in BTV2 infected PBMC. 54 Figure 6. Proposed pathogenesis and immnity between BTV2 and bovine PBMC 55 Table 1. Cytokine profiles to various BTV strains 56 Table 2. Primer sequences for the quantification of cytokine and viral genome expression by one step qRT-PCR 58 | |
| dc.language.iso | en | |
| dc.subject | 致病機轉 | zh_TW |
| dc.subject | 藍舌病 | zh_TW |
| dc.subject | 周邊血液單核球 | zh_TW |
| dc.subject | 細胞激素 | zh_TW |
| dc.subject | 信使核糖核酸表現 | zh_TW |
| dc.subject | mRNA expression | en |
| dc.subject | pathogenesis | en |
| dc.subject | cytokines | en |
| dc.subject | peripheral mononuclear blood cell | en |
| dc.subject | Bluetongue virus | en |
| dc.title | 臺灣藍舌病病毒接種牛隻周邊血液單核球之細胞激素分析 | zh_TW |
| dc.title | Cytokine Profile of Bovine Peripheral Blood Mononuclear Cells Challenged Against Low Virulent Bluetongue Virus Taiwan Isolate | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 106-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 李龍湖,許天來,李璠,張家宜 | |
| dc.subject.keyword | 藍舌病,周邊血液單核球,細胞激素,信使核糖核酸表現,致病機轉, | zh_TW |
| dc.subject.keyword | Bluetongue virus,peripheral mononuclear blood cell,cytokines,mRNA expression,pathogenesis, | en |
| dc.relation.page | 62 | |
| dc.identifier.doi | 10.6342/NTU201803890 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2018-08-18 | |
| dc.contributor.author-college | 獸醫專業學院 | zh_TW |
| dc.contributor.author-dept | 分子暨比較病理生物學研究所 | zh_TW |
| dc.date.embargo-lift | 2023-08-21 | - |
| 顯示於系所單位： | 分子暨比較病理生物學研究所 | |
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