Subproject 1: Virus-host interaction involved in inter-species barrier
This subproject will identify viral genetic determinants and cellular mechanisms of influenza inter-species barrier at the nuclear export process in human cells by:
1. Identifying the genetic determinant of nuclear export defect in human cells of an H7N1 avian influenza virus using reverse genetic techniques, reassortment and site-specific mutagenesis.
2. Indentifying the species-specific interaction between the viral nuclear export protein and cellular nuclear export machinery using co-immunoprecipitation.
Subproject 2: Virus-host interaction in interferon evasion
1. To study the role of heat shock proteins in the mechanism of interferon evasion by influenza virus in the cold.
2. To study the role of poly(rC)-binding protein (PCBP) and the viral protein 3C in the interferon evasion by enterovirus 71.
Subproject 3: Risk evaluation of emerging zoonotic viruses using pseudotyped viruses
1. To develop pseudotyped viral reporter vector carrying surface glycoproteins of newly identified viruses of interest.
2. To obtain primary cells or cell lines of respiratory and gastrointestinal origin from bat, rodent, pig, monkey and human.
3. To study relative transducing ability of the pseudotyped vector in those cell culture.
4. To evaluate the mutations required to enhance the transduction in human cells.
Subproject 4: Epitope characterization of antibody response to viral infection
1. To study the kinetics of antibody to the stalk domain of hemagglutinin of the 2009 H1N1 pandemic influenza virus in general population.
2. To characterize epitopes recognized by primary and secondary antibody responses to dengue infection.