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標題: 建立氣管上皮細胞細胞株以利家禽呼吸道病毒的鑑定與增殖
Establishment of Cell Lines of Tracheal Epithelial Cells for the Identification and Amplification of Avian Respiratory Viruses
作者: 蘇鴻麟
關鍵字: 生物技術;基礎研究
目前禽類呼吸道病毒多於雞胚或雞纖維母細胞中增殖,此兩種繼代法的缺點分別有,病毒因需適應新宿主細胞常產生基因突變,以及所產生的病毒力價過低。此外,部份呼吸道病毒的毒力鑑定,如家禽支氣管炎病毒,因大都對纖維母細胞無細胞毒性,目前主要靠雞胚的生長停滯與氣管環上的纖毛擺動速率來認定,也有無法準確測出病毒力價與毒力的缺點。為解決以上問題,本計畫將藉由已建立之家禽氣管上皮細胞初代培養技術,利用腺病毒(adenovirus)與Nucleofactor,將致癌基因large T antigen, H-Ras以及c-myc送入細胞中,建立新的禽類呼吸道上皮細胞株。由於纖維母細胞容易感染慢病毒lentivirus,而呼吸道上皮細胞則否。我們將利用lentivirus帶DT-A (diphtheria toxin A),去除繼代過程中的纖維母細胞。建立的細胞株將可提供一新穎、簡易且高效率培養禽類病毒的增殖平台。利用此初代培養細胞與所建立之各種細胞株,並可研究呼吸道病毒如何感染上皮細胞的機制。所建立的細胞株除可提供野外分離病毒之增殖與鑑定外,我們也將評估是否此細胞株可提供一穩定、低成本與高效率的流行性感冒病毒疫苗株的製造平台。

In the present, most avian respiratory viruses are amplified in the chorioallantoicmembrane of embryonated eggs or chicken fibroblast cells. The viral adaptation to the newhost cells often associates with genetic mutation and low viral production. In addition, mostinfectious bronchitis viruses are non-cytopathic to fibroblast cells and their viral titers andvirulences can only be roughly estimated by the growth retention of embryos and ciliostasis oftracheal rings. To tackle these problems, our primary tracheal epithelial cell system will beapplied to establish the stable cell lines of bronchial epithelium, combined with the infectionof the recombinant adenovirus carrying the SV40 large T antigen, H-Ras and c-myconcogenes. We found that recombinant lentivirus showed high cell tropism to the fibroblastbut not to the respiratory epithelial cells. Infection with the diphtheria toxin A (DTA)recombinant lentivirus should reduce the growth of fibroblast cells and their contamination inthe transformed cell population. Primary tracheal epithelial cells and their derivative cell lineswill benefit the understanding of the mechanism of viral infection. Established cell lines willprovide a novel system for the detection, isolation and amplification of the avian viruses. Inaddition, the stable cell line may also be a stable and high efficient factory for the productionof vaccine strains of avian influenza viruses.
其他識別: NSC99-2321-B005-012-MY3
Appears in Collections:生命科學系所

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