title: Cell penetrable human scFv specific to middle domain of matrix protein-1 protects mice from lethal influenza.
abstract: A new anti-influenza remedy that can tolerate the virus antigenic variation is needed. Influenza virus matrix protein-1 (M1) is highly conserved and pivotal for the virus replication cycle: virus uncoating, assembly and budding. An agent that blocks the M1 functions should be an effective anti-influenza agent. In this study, human scFv that bound to recombinant M1 middle domain (MD) and native M1 of A/H5N1 was produced. Phage mimotope search and computerized molecular docking revealed that the scFv bound to the MD conformational epitope formed by juxtaposed helices 7 and 9 of the M1. The scFv was linked molecularly to a cell penetrable peptide, penetratin (PEN). The PEN-scFv (transbody), when used to treat the cells pre-infected with the heterologous clade/subclade A/H5N1 reduced the viral mRNA intracellularly and in the cell culture fluids. The transbody mitigated symptom severity and lung histopathology of the H5N1 infected mice and caused reduction of virus antigen in the tissues as well as extricated the animals from the lethal challenge in a dose dependent manner. The transbody specific to the M1 MD, either alone or in combination with the cognate human scFvs specific to other influenza virus proteins, should be an effective, safe and mutation tolerable anti-influenza agent.
authors.affiliation: Center for Agricultural Biotechnology, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand. fddvt66@outlook.com.
authors.forename: Fonthip
authors.initials: F
authors.lastname: Dong-din-on
authors.affiliation: Department of Veterinary Pathology, Faculty of Veterinary Medicine, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand. fvettss@ku.ac.th.
authors.forename: Thaweesak
authors.initials: T
authors.lastname: Songserm
authors.affiliation: Laboratory for Research and Technology Development, Department of Parasitology, and Center of Excellence on Therapeutic Proteins and Antibody Engineering, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. tpw029@gmail.com.
authors.forename: Tippawan
authors.initials: T
authors.lastname: Pissawong
authors.affiliation: Graduate Program in Biomedical Science, Faculty of Allied Health Sciences, Thammasat University, Pathumthani 12120, Thailand. psrimanote01@yahoo.com.au.
authors.forename: Potjanee
authors.initials: P
authors.lastname: Srimanote
authors.affiliation: Laboratory for Research and Technology Development, Department of Parasitology, and Center of Excellence on Therapeutic Proteins and Antibody Engineering, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. psrimanote01@yahoo.com.au.
authors.forename: Jeeraphong
authors.initials: J
authors.lastname: Thanongsaksrikul
authors.affiliation: Laboratory for Research and Technology Development, Department of Parasitology, and Center of Excellence on Therapeutic Proteins and Antibody Engineering, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. mam_mt41@hotmail.com.
authors.forename: Kanyarat
authors.initials: K
authors.lastname: Thueng-in
authors.affiliation: Department of Veterinary Pathology, Faculty of Veterinary Medicine, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand. fvetpas@ku.ac.th.
authors.forename: Pattra
authors.initials: P
authors.lastname: Moonjit
authors.affiliation: Department of Veterinary Pathology, Faculty of Veterinary Medicine, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand. preeda_l@ku.ac.th.
authors.forename: Preeda
authors.initials: P
authors.lastname: Lertwatcharasarakul
authors.affiliation: Laboratory for Research and Technology Development, Department of Parasitology, and Center of Excellence on Therapeutic Proteins and Antibody Engineering, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. watee.see@gmail.com.
authors.forename: Watee
authors.initials: W
authors.lastname: Seesuay
authors.affiliation: Laboratory for Research and Technology Development, Department of Parasitology, and Center of Excellence on Therapeutic Proteins and Antibody Engineering, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. wanpen.cha@mahidol.ac.th.
authors.forename: Wanpen
authors.initials: W
authors.lastname: Chaicumpa
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics / Q000627:therapeutic use*; D000998:Antiviral Agents / Q000627:therapeutic use*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D057846:Cell-Penetrating Peptides; D004195:Disease Models, Animal; D005260:Female; D053124:Influenza A Virus, H5N1 Subtype / Q000187:drug effects*; D008807:Mice, Inbred BALB C; D062105:Molecular Docking Simulation; D009976:Orthomyxoviridae Infections / Q000188:drug therapy* / Q000473:pathology / Q000821:virology; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D057127:Single-Chain Antibodies / Q000235:genetics / Q000627:therapeutic use*; D016019:Survival Analysis; D016896:Treatment Outcome; D014763:Viral Matrix Proteins / Q000037:antagonists & inhibitors*
chemical_list: D000914:Antibodies, Viral; D000998:Antiviral Agents; D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; C488936:M1 protein, Influenza A virus; D019151:Peptide Library; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D014763:Viral Matrix Proteins; C414409:penetratin