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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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Histopathological features in lung sections of mice infected with 10 MLD50 of mouse adapted A/chicken/Thailand/NP-172/2006 (H5N1) at 96 h post infection in comparison with the lung section of a normal mouse. (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 (H & E; 20× original magnification).
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viruses-07-00154-f006: Histopathological features in lung sections of mice infected with 10 MLD50 of mouse adapted A/chicken/Thailand/NP-172/2006 (H5N1) at 96 h post infection in comparison with the lung section of a normal mouse. (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 (H & E; 20× original magnification).

Mentions: Lung histopathological features of the infected mice that received different treatments at 96 h p.i., in comparison with the histology of the lung of a normal mouse are shown in Figure 6. The untreated infected mice (PBS control group) showed the most severe pulmonary lesion including multifocal necrotizing lymphocytic-neutrophilic interstitial pneumonia and necrosis of the lung tissue. Cellular debris (admixed desquamated epithelial cells and degenerative neutrophils) was found in the bronchiolar and alveolar spaces (Figure 6I). The increasing order of the pathological degrees in the lungs of the treated infected mice were PAb to H5N1 (Figure 6G) = rimantadine (Figure 6H) < PEN-scFv21 (Figure 6B) < scFv21 (Figure 6A) < control scFv (Figure 6C) = control PEN-scFv (Figure 6D) = E. coli lysate (Figure 6E) = PAb to M1 (Figure 6F).


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)

Histopathological features in lung sections of mice infected with 10 MLD50 of mouse adapted A/chicken/Thailand/NP-172/2006 (H5N1) at 96 h post infection in comparison with the lung section of a normal mouse. (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 (H & E; 20× original magnification).
© Copyright Policy
Related In: Results  -  Collection

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

viruses-07-00154-f006: Histopathological features in lung sections of mice infected with 10 MLD50 of mouse adapted A/chicken/Thailand/NP-172/2006 (H5N1) at 96 h post infection in comparison with the lung section of a normal mouse. (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 (H & E; 20× original magnification).
Mentions: Lung histopathological features of the infected mice that received different treatments at 96 h p.i., in comparison with the histology of the lung of a normal mouse are shown in Figure 6. The untreated infected mice (PBS control group) showed the most severe pulmonary lesion including multifocal necrotizing lymphocytic-neutrophilic interstitial pneumonia and necrosis of the lung tissue. Cellular debris (admixed desquamated epithelial cells and degenerative neutrophils) was found in the bronchiolar and alveolar spaces (Figure 6I). The increasing order of the pathological degrees in the lungs of the treated infected mice were PAb to H5N1 (Figure 6G) = rimantadine (Figure 6H) < PEN-scFv21 (Figure 6B) < scFv21 (Figure 6A) < control scFv (Figure 6C) = control PEN-scFv (Figure 6D) = E. coli lysate (Figure 6E) = PAb to M1 (Figure 6F).

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