GOTO Hideo The University of Tokyo, Institute of Medical Science, Research Assistant, 医科学研究所, 助手 (50323639)
TAKADA Ayato The University of Tokyo, Institute of Medical Science, Research Assistant, 医科学研究所, 助手 (10292062)
|Budget Amount *help
¥2,900,000 (Direct Cost : ¥2,900,000)
Fiscal Year 2003 : ¥1,100,000 (Direct Cost : ¥1,100,000)
Fiscal Year 2002 : ¥1,800,000 (Direct Cost : ¥1,800,000)
Progeny viruses are rapidly produced from the cells infected with influenza A viruses for a short period. A several cellular factors regulate virus replication in cells, interacting with viral proteins or RNAs directly or indirectly. In this study, we sought to identify and analyze cellular factors responsible for virus replication. It is hopeful that such information would contribute one to develop novel anti-influenza drugs in the future. To this end, we attempted to establish and use new methodology to identify cellular factors ; that is, "self-motivating" recombinant virus system. In this system, we constructed recombinant influenza viruses, such as HA-NA tandem construct or 9 segment construct, randomly accommodating cellular cDNAs derived from virus-susceptible tissues and cells. Theoretically, if cellular factors responsible for virus replication are expressed by these cDNAs, recombinant viruses would replicate even in virus-insusceptible cells as well. In this case, we will be able to identify cellular factors responsible for virus replication by cDNA sequencing in each recombinant virus. First, we tried to select cell clones insensitive to influenza virus infection by treatment of Chinese hamster ovary CHO cells with chemical mutagenesis reagent ICR191, and established a total of 72 clones. On the other hand, we constructed recombinant viruses accommodating cDNAs prepared from human lung tissues by plasmid-based reverse genetics protocols. Then, we infected each cell clone with these recombinant viruses. However, we were not able to obtain infectious viruses replicating in these cell clones. Thus, we have not identified cellular factors responsible for virus replication so far. We are not sure whether our recombinant virus system did not work, or whether our selected cell clone insensitive to virus infection was not suitable for this study.