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標題: 研發可同時表現日本腦炎病毒封套蛋白及豬介白素-2之假性狂犬病重組病毒疫苗
Development of a Recombinant Pseudorabies Virus Co-Expressing JEV Envelope Protein and Swine IL-2
作者: 張天傑
關鍵字: 應用研究;pseudorabies virus (PRV);生物技術, 畜牧獸醫類;假性狂犬病毒;日本腦炎病毒;封套蛋白E;TK;gE;vhs;UL41基因;豬介白素-2;重組病毒;Japanese encephalitis virus (JEV);E protein;TK;gE;vhs;sIL-2;UL41 gene;recombinant virus
假性狂犬病毒(pseudorabies virus, PRV)是疱疹病毒科的阿爾伐疱疹病毒亞科內之一員,可以感染許多的哺乳類動物,在台灣是重要的猪隻傳染病之一,影響本省農業經濟甚鉅。日本腦炎係由日本腦炎病毒(Japanese encephalitis virus, JEV)感染引起的急性腦膜腦炎,此病為一種病毒性人畜共通疾病,其感染途徑是藉由三斑家蚊與環紋家蚊等病媒蚊傳染。妊娠母猪感染日本腦炎後,胎盤受病毒侵害,引起流產和死產。台灣夏季常有孩童感染腦炎症的病例,本病在公共衛生上是不可忽視的重要疾病。而使養猪界煩惱的猪的死產及流產,主要是由於猪感染日本腦炎病毒或假性狂犬病毒所致。目前全世界已有推行假性狂犬病撲滅計劃的趨勢,因此,良好的疫苗研發,將是控制這些重要病毒性疾病的重要利器。本實驗室已完成了PRV的vhs缺損病毒及R23 (TK-/gE-)病毒的選殖及特性分析,根據in vitro及in vivo的實驗結果發現,vhs的刪除並不會影響PRV的複製生長,且其毒力也有明顯下降的趨勢。同時也發現vhs缺陷病毒感染細胞或小鼠後,能夠測到大量的TNF-α;且能延長小鼠的死亡時間及存活率,其臨床的感染症狀也比較輕微。因為TK、gE及vhs都和PRV的病毒毒力有關;但並不會直接影響PRV的複製;所以將這3種基因刪除後,會降低PRV的毒力使其成為一個安全的病毒載體。同時我們也完成了猪的IL-2及日本腦炎病毒的套膜蛋白E基因之選殖,也能利用原核系統表現出相對應之蛋白質。而套膜蛋白E具有3個抗原決定位,其中以第三個抗原決定位(ED3)為3個抗原決定區域中唯一能單獨折疊的抗原區域,可誘發宿主產生病毒專一性中和抗體反應及保護性免疫反應的產生。因此,我們擬將猪的介白素-2(IL-2)及JEV封套蛋白E的第三個抗原決定位E-D3基因一起植入假性狂犬病毒之基因體內,以開發具有佐劑特質及抗假性狂犬病與日本腦炎之多價活毒疫苗。

Pseudorabies, caused by the alphaherpesvirus pseudorabies virus (PRV), is a serious infection in a wide range of mammals. Pig is the natural reservoir for the virus, and infection with PRV results reproductive and respiratory disorders, nervous symptoms and even death, dependent on the age of the animal and virus strain-specific virulence. Japanese encephalitis is a potentially lethal disease of the central nervous system caused by infection with Japanese encephalitis virus (JEV), a member of the mosquito-borne encephalitis complex of the family Flaviviridae. Swine are an important amplifier of Japanese encephalitis (JE) virus and PRV in the paradomestic environment. PRV and Japanese encephalitis virus (JEV) are major global health and growing medical problems. When piglets are infected with PRV or JEV have caused strillbirths and abortions in pregnant sows and cause significant losses in the pig industry worldwide. To date, eradication of PRV and control of JEV become important tasks in the pig industry to reduce economical losses. From our previous study, we constructed and characterized that R23 (gE-/TK-) and vhs mutant virus (Δ41). The inactivation of TK and gE synergistically reduces PRV replication and virulence in vivo. While the vhs mutant virus has similar growth kinetics to wild type PRV, absence of vhs activity significantly reduced PRV virulence; wild type is lethal, whereas vhs mutant is highly attenuated in mice model. Moreover, we found the lethality is correlated with the dissemination of virus in neural tissues where only wt PRV but not vhs deletion virus was detected. Hence, the gene-delected of TK, gE and vhs in PRV has been used as a live-viral vector to develop multivalent genetic engineering vaccine. The E protein of JEV contains 3 antigenic domains and plays important roles in receptor binding and cell fusion and possesses most of the virus-neutralizing epitopes. The major antigenic domain of E protein is domain III (ED3). In this study, we will develope and test a novel recombinant PRV, which could co-express protein of the swine IL-2 and ED3 protein of JEV using PRV TK-/gE- mutant (R23) as the vector. The immunogenicity of this recombinant virus will be compared to the currently used commercial JEV and PRV vaccine in animal model.
其他識別: NSC99-2313-B005-007-MY3
Appears in Collections:獸醫學系所

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