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Title: | 豬鐵士古病毒攜帶口蹄疫病毒VP1殼蛋白及豬瘟E2醣蛋白抗原決定位作為疫苗載體的可行性 Feasibility of Porcine Teschovirus Derived Vector carrying both Foot and Mouth Disease VP1 capsid protein and Classical Swine Fever E2 glycoprotein epitopes as a Vaccine |
Authors: | Chih-Shuan Sheng 沈芷萱 |
Advisor: | 王汎熒 |
Keyword: | 豬鐵士古病毒,病毒源性載體,免疫螢光試驗, porcine teschovirus,virus-derived vector,immunofluorescent assay, |
Publication Year : | 2018 |
Degree: | 碩士 |
Abstract: | 小核糖核酸病毒科(Picornaviridae)是一個包含大量動物致病原的病毒科,也是用於研究正向單股RNA病毒生物學、發病機轉和流行病學的模型病毒,近年來反向遺傳學技術的發展,使得此病毒的結構和功能得以詳細分析,因而可作為疫苗載體的候選病毒之一。例如:脊髓灰質炎病毒 (Poliovirus) 具有作為疫苗載體的優點,包括易於口服遞送,腸道內保持穩定性,以及誘導體液和細胞免疫的能力。本研究中豬鐵士古病毒(Porcine Teschovirus, PTV)相似於脊髓灰質炎病毒,亦有其開發疫苗載體的優勢,且不是OIE表列之重大動物傳染病病毒。雖在全球的豬群中廣泛存在,但均為散發流行的弱毒株。因此將其作為疫苗載體可降低環境衝擊和疾病預防的風險。此外,PTV具有廣泛的組織親和性,使其可作為攜帶外源基因開發多價黏膜疫苗載體的平台。PTV的基因體較短,長度為7~9 kb,因此更容易構築以表現外源蛋白抗原決定位。本研究利用豬瘟病毒E2蛋白和口蹄疫病毒VP1蛋白抗原決定位(epitope) 取代PTV病毒VP1蛋白上的BC環狀區域構築重組載體,並使用poly-His tag替代G-H環狀區域以區分野外株病毒或進行蛋白質純化和檢測之用。結果顯示,使用單株和多株抗體進行免疫螢光檢測,重組PTV載體能夠表現兩種外源蛋白抗原決定位,證明其作為疫苗平台之潛力。 The family Picornaviridae comprises a large number of animal pathogens, and it is also a model system for the study of positive single-stranded RNA virus biology, pathogenesis, and epidemiology. The development of reverse genetics technology allows the analysis in details of the structure and function of these viruses. Poliovirus has several advantages to serve as vaccine vector, including easy to deliver orally, inherent stability in the intestinal tract, and the ability to induce both humoral and cell immunities. Like poliovirus, porcine teschovirus (PTV) belongs to the family Picornaviridae, which is a positive single-strand RNA virus. The aim of this study was to take the advantages of PTV to develop a vaccine vector. PTV encephalomyelitis is not an OIE listed disease that could be widely detected in swine herds worldwide, and yet shows mild virulence (except for a few recent virgin epidemics), thus lowers the risk of environmental impact and disease prevention by using it as a vaccine vector. Also, PTV has a wide tissue tropism making it feasible to be used as a platform to develop multivalent mucosal vaccine vector by carrying heterogenous genes. Its genome length of only 7 - 9 kb makes it easier to construct and to present heterogenous proteins. Thus, a recombinant PTV vector was constructed by replacing the B-C loop region on its VP1 protein with both classical swine fever virus (CSFV) E2 and foot-and-mouth disease (FMDV) VP1 epitopes, and its G-H loop region by poly-His tag for easier distinction from wildtype virus and easier protein purification and detection if necessary. Immunofluorescent assay showed that the constructed PTV vector was able to express both heterogenous epitopes, that could be detected by both polyclonal and monoclonal antibodies directed to each protein or antigenic sites. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78923 |
DOI: | 10.6342/NTU201802334 |
Fulltext Rights: | 有償授權 |
metadata.dc.date.embargo-lift: | 2023-08-23 |
Appears in Collections: | 分子暨比較病理生物學研究所 |
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ntu-107-R05644002-1.pdf Restricted Access | 1.99 MB | Adobe PDF |
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