A human single chain transbody specific to matrix protein (M1) interferes with the replication of influenza A virus.
A cell penetrating format of human single chain antibody (HuScFv) specific to matrix protein (M1) of influenza A virus was produced by molecular linking of the gene sequence encoding the HuScFv (huscfv) to a protein transduction domain, i.e., penetratin (PEN) of the Drosophila homeodomain. DNA of a recombinant phagemid vector carrying the huscfv was used as a platform template in a three-step PCR for generating a nucleotide sequence encoding a 16 amino acid PEN peptide. The PEN-HuScFv had negligible cytotoxicity on living MDCK cells. They were readily translocated across the cell membrane and bound to native M1 in the A/H5N1-infected cells as revealed by immunofluorescent confocal microscopy. The PEN-HuScFv, when used to treat the influenza virus infected cells, reduced the number of viruses released from the cells. In conclusion, the cell penetrating M1-specific HuScFv, a transbody, produced in this study affected the influenza A virus life cycle in living mammalian cells. While the molecular mechanisms of the PEN-HuScFv need more investigation, the reagent warrants further testing in animals before developing it into a human immunotherapeutic anti-influenza formula.
Poungpair
Ornnuthchar
O
Faculty of Allied Health Sciences, Thammasat University, Rangsit Center, Pathumthani 12120, Thailand.
Pootong
Anek
A
Maneewatch
Santi
S
Srimanote
Potjanee
P
Tongtawe
Pongsri
P
Songserm
Thaweesak
T
Tapchaisri
Pramuan
P
Chaicumpa
Wanpen
W
D000918:Antibody Specificity; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D057846:Cell-Penetrating Peptides; D006801:Humans; D009980:Influenza A virus / Q000187:drug effects* / Q000254:growth & development / Q000276:immunology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology*; D014763:Viral Matrix Proteins / Q000276:immunology*; D014779:Virus Replication / Q000187:drug effects*
D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D057127:Single-Chain Antibodies; D014763:Viral Matrix Proteins; C414409:penetratin