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Cell penetrable human scFv specific to middle domain of matrix protein-1 protects mice from lethal influenza.

Dong-din-on F, Songserm T, Pissawong T, Srimanote P, Thanongsaksrikul J, Thueng-in K, Moonjit P, Lertwatcharasarakul P, Seesuay W, Chaicumpa W - Viruses (2015)

Bottom Line: An agent that blocks the M1 functions should be an effective anti-influenza agent.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 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.

View Article: PubMed Central - PubMed

Affiliation: Center for Agricultural Biotechnology, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand. fddvt66@outlook.com.

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.

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Related in: MedlinePlus

Immunohistochemical staining for influenza virus nucleoprotein (brownish gold pigments) in lung sections (40× original magnification) of mice infected with 10 MLD50 of mouse adapted A/chicken/Thailand/NP-172/2006 (H5N1), clade 2, subclade 3 at 96 h post infection. (A–H) Infected mice treated with 10 mg/kg body weight of scFv21, PEN-scFv21, control scFv, control PEN-scFv, E. coli lysate, PAb to M1, PAb to H5N1 and rimantadine; (I) Infected mouse treated with PBS and (J) normal mouse.
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viruses-07-00154-f007: Immunohistochemical staining for influenza virus nucleoprotein (brownish gold pigments) in lung sections (40× original magnification) of mice infected with 10 MLD50 of mouse adapted A/chicken/Thailand/NP-172/2006 (H5N1), clade 2, subclade 3 at 96 h post infection. (A–H) Infected mice treated with 10 mg/kg body weight of scFv21, PEN-scFv21, control scFv, control PEN-scFv, E. coli lysate, PAb to M1, PAb to H5N1 and rimantadine; (I) Infected mouse treated with PBS and (J) normal mouse.

Mentions: Lung sections of mice stained for the influenza virus nucleoprotein (NP) are shown in Figure 7. Negligible amount of the NP was observed in lung sections of infected mice treated with PAb to H5N1 (Figure 7G) and rimantadine (Figure 7H) in comparison with the sections of the PBS treated infected mice (Figure 7I) which revealed the most of the NP among all infected mouse groups. There was much less NP in the lung sections of mice treated with PEN-scFv21 (Figure 7B) and scFv21 (Figure 7A) compared with the PBS group. Although the amounts of NP in lungs of the groups treated with control scFv (Figure 7C), control PEN-scFv (Figure 7D), E. coli lysate (Figure 7E) and PAb to M1 (Figure 7F) were slightly less than the PBS group, they were much more than the PAb to H5N1, the rimantadine, the PEN-scFv and the scFv treated groups.


Cell penetrable human scFv specific to middle domain of matrix protein-1 protects mice from lethal influenza.

Dong-din-on F, Songserm T, Pissawong T, Srimanote P, Thanongsaksrikul J, Thueng-in K, Moonjit P, Lertwatcharasarakul P, Seesuay W, Chaicumpa W - Viruses (2015)

Immunohistochemical staining for influenza virus nucleoprotein (brownish gold pigments) in lung sections (40× original magnification) of mice infected with 10 MLD50 of mouse adapted A/chicken/Thailand/NP-172/2006 (H5N1), clade 2, subclade 3 at 96 h post infection. (A–H) Infected mice treated with 10 mg/kg body weight of scFv21, PEN-scFv21, control scFv, control PEN-scFv, E. coli lysate, PAb to M1, PAb to H5N1 and rimantadine; (I) Infected mouse treated with PBS and (J) normal mouse.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4306832&req=5

viruses-07-00154-f007: Immunohistochemical staining for influenza virus nucleoprotein (brownish gold pigments) in lung sections (40× original magnification) of mice infected with 10 MLD50 of mouse adapted A/chicken/Thailand/NP-172/2006 (H5N1), clade 2, subclade 3 at 96 h post infection. (A–H) Infected mice treated with 10 mg/kg body weight of scFv21, PEN-scFv21, control scFv, control PEN-scFv, E. coli lysate, PAb to M1, PAb to H5N1 and rimantadine; (I) Infected mouse treated with PBS and (J) normal mouse.
Mentions: Lung sections of mice stained for the influenza virus nucleoprotein (NP) are shown in Figure 7. Negligible amount of the NP was observed in lung sections of infected mice treated with PAb to H5N1 (Figure 7G) and rimantadine (Figure 7H) in comparison with the sections of the PBS treated infected mice (Figure 7I) which revealed the most of the NP among all infected mouse groups. There was much less NP in the lung sections of mice treated with PEN-scFv21 (Figure 7B) and scFv21 (Figure 7A) compared with the PBS group. Although the amounts of NP in lungs of the groups treated with control scFv (Figure 7C), control PEN-scFv (Figure 7D), E. coli lysate (Figure 7E) and PAb to M1 (Figure 7F) were slightly less than the PBS group, they were much more than the PAb to H5N1, the rimantadine, the PEN-scFv and the scFv treated groups.

Bottom Line: An agent that blocks the M1 functions should be an effective anti-influenza agent.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 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.

View Article: PubMed Central - PubMed

Affiliation: Center for Agricultural Biotechnology, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand. fddvt66@outlook.com.

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.

Show MeSH
Related in: MedlinePlus