Study on antisense phosphorothioate oligodeoxynudeotide resistance to influenza A virus infection in vitro

Abstract

Objective To investigate the antiviral effects of antisense phosphorothioate oligodeoxynucleotide (ASODN)in 9HTEO infected with influenza A virus (IFAV) in vitro. Methods The ASODN which complemented to genomic PB1, M2, NS, PB2, HA mRNA of IFAV was used to investigate antiviral effection in vitro. The cytopathic effect (CPE) was observed, and the cell survival rates were measured by MTF assay, plaque assay, RT-PCR, Western blot and Immunofluorescence were performed to test anti-viral efficiency of PB1, M2, NS in cells mRNA and protein level. Results 9HTEO cells infected with IFAV almost all died when the multipicity of infection (MOI) is aboved after 5 days cell culture. The ASODN could increase the cell survival rates. The IFAV PB1, M2, NS significantly reduced CPE of IFAV infected 9HTEO cells, reduced the viral replication of IFAV in the cells (P <0.05). Conclusion The ASODN which targeted the mRNA of IFAV gene showed a significant and specific anti-IFAV effect both in mRNA and protein level in cells culture system. The study indicates that the PB1, M2 and NS mRNA may play an important role in regulating IFAV replication, and ASODN may have inhibitory activity on IFAV replication. The results established the basis for further study on new drugs against IFAV infection include the highly pathogenic H5N1 influenza virus. Key words: Influenza A virus; Antisense oligodeoxynucleotide; 9HTEO cell; Virus replication