| Summary: | 碩士 === 慈濟大學 === 醫學生物技術研究所 === 98 === “Flu” is a contagious respiratory illness mainly caused by influenza A virus. Flu outbreaks due to emerging influenza A virus infections could cause millions death, e.g. 1918 Spanish flu (H1N1), 1957 Asian flu (H2N2) and 1968 Hong Kong flu (H3N2).
To search for cellular factors modulating influenza A virus replication, microarray analysis was performed to detect the cellular genes differentially expressed after influenza A virus WSN33 infection in A549 cells at different time periods (2 hrs, 4 hrs, 23 hrs). As expected, cellular genes related to immune responses were modulated after virus infection. Interestingly, various histone genes were up-regulated after virus infection. Up-regulation of histone was confirmed using real time RT-PCR to detect mRNA level and Western blotting analysis to detect protein amount. To determine the effect of up-regulated histone gene on influenza A virus replication, siRNA technology was used to knock-down histone gene expression in A549 cells. Reduction of histone gene expression was confirmed using Western blotting analysis to detect protein amount. Western blotting analysis to detect viral protein expression and plague assay to analyze the viral particles were used to examine the replication of influenza A virus. Our results showed that reduction of histone expression did inhibit virus replication.
When influenza A virus NS1 gene was transfected to A549 cells, the histone expression was not increased. Interestingly, histones were found in the viral particles. Molecular mechanisms regarding how influenza A virus up-regulates histone gene expression and how histone affects virus replication will be addressed in the future.
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