Cell penetrable human ScFv specific to influenza A virus matrix protein, M1, mitigates influenza severity

Abstract

Event Abstract Back to Event Cell penetrable human ScFv specific to influenza A virus matrix protein, M1, mitigates influenza severity Fonthip Dong-din-on1, Potjanee Srimanote2, Tippawan Pissawong3, Angkasiya Monkong4, Preeda Lertwatcharasarakul4, Thaweesak Songserm4 and Wanpen Chaicumpa5* 1 Kasetsart University, Center for Agricultural Biotechnology, Thailand 2 Thammasat University, Graduate Program in Biomedical Science, Faculty of Allied Health Sciences, Thailand 3 Mahidol University, Department of Immunology, Faculty of Medicine Siriraj Hospital, Thailand 4 Kasetsart University, Veterinary Midicine, Thailand 5 Mahidol University, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Thailand Matrix protein-1 (M1) is highly conserved across type A influenza viruses. This protein has many important functions in the viral replication cycle; thus it is one of the targets of novel anti-influenza. To study therapeutic efficacy of M1 specific, cell penetrable human single chain antibody (HuScFv) in mitigating severity of influenza in infected mice. E. coli derived-human single chain antibody fragments (HuScFv) specific to recombinant M1 of influenza A virus H5N1 (clade 1) was produced using a human antibody phage display library. HuScFv from selected huscfv-phagemid transformed E. coli clones were linked molecularly to a cell penetrating peptide, penetratin (PEN) and PEN-HuScFv were produced and purified. BALB/c mice were infected intranasally with mouse adapted-avian H5N1 virus (clade 2.3). They were then treated with M1 specific-PEN-HuScFv. Control infected mice received PBS treatment. Internal organs (lung, brain, spleen, liver and kidney) were collected. Tissue viral loads were determined by real time RT-PCR. Histopathology of the tissues was also examined. Infected BALB/c mice that received M1 specific-PEN-HuScFv had reduced viral loads and histopathological features in tissues compared to the controls. M1 specific-PEN-HuScFv could mitigate severity of influenza in mice infected with A H5N1 of the heterologous clade. Cross therapeutic efficacy of the transbodies on influenza caused by different virus subtypes remains to be evaluated. Acknowledgements This research work was financially supported by grant no. DPG5380001 of the Thailand Research Fund (TRF) and the National Research University (NRU) Project, Office of Higher Education Commission (CHE), Ministry of Education, Thailand. Fonthip is a TRF scholar of the Royal Golden Jubilee Ph.D. Program.