| Summary: | 碩士 === 國立臺灣大學 === 漁業科學研究所 === 88 === Abstract
IPNV (Infectious Pancreatic Necrosis Virus) is a double-stranded RNA viruse which has been shown to be a major cause of many contagious and widespread fish diseases. IPNV belongs to Birnaviridae virus family and is also a pathogen of many economically important fishes in Taiwan. The IPNV-E1S (Eel No.1 Spleen) virus strain was isolated from the spleen of diseased Japanese eel (Anguilla japonica) and is an Ab serotype of Aquatic Birnavirus. Recent study has established apoptosis as a major cause for the observed IPNV-induced cytopathic effects in infected fish cells. IPNV-infected CHSE-214 cells displayed all previously known morphological and biochemical hallmarks of apoptosis. At present, the apoptosis-inducing activity of IPNV has not been definitively assigned to any IPNV factors. Nevertheless, there was evidence implicating VP3 as such a factor. To identify the unknown apoptosis-inducing element of IPNV, CHSE-214 cells stably expressing various antisense RNAs (A2, A4, and A7, complement to nt 501-711, 285-711, and 1-711, respectively) against VP3 were established. All antisense RNA expressing cells showed increased resistance to IPNV-induced apoptosis in viral challenge experiments (MOI 1). Further analyses showed that VP3 protein expression was suppressed, Mcl-1 expression was longer sustained, and DNA fragmentation was reduced in these antisense RNA expressing cells during early viral replication cycle. These results suggest that the antisense RNAs could knock down the VP3 expression and VP3 is an apoptosis-inducing factor responsible for the IPNV-induced apoptotic cell death.
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