Limits...
An Anti-Influenza Virus Antibody Inhibits Viral Infection by Reducing Nucleus Entry of Influenza Nucleoprotein.

Yoon A, Yi KS, Chang SY, Kim SH, Song M, Choi JA, Bourgeois M, Hossain MJ, Chen LM, Donis RO, Kim H, Lee Y, Hwang do B, Min JY, Chang SJ, Chung J - PLoS ONE (2015)

Bottom Line: To date, four main mechanisms mediating inhibition of influenza infection by anti-hemagglutinin antibodies have been reported.In this study we identified a human monoclonal IgG1 antibody (CT302), which does not inhibit both the receptor binding and the membrane fusion process but efficiently reduced the nucleus entry of viral nucleoprotein suggesting a novel inhibition mechanism of viral infection by antibody.This antibody binds to the subtype-H3 hemagglutinin globular head domain of group-2 influenza viruses circulating throughout the population between 1997 and 2007.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul National University, Seoul, South Korea; Cancer Research Institute, Seoul National University College of Medicine, Seoul National University, Seoul, South Korea.

ABSTRACT
To date, four main mechanisms mediating inhibition of influenza infection by anti-hemagglutinin antibodies have been reported. Anti-globular-head-domain antibodies block either influenza virus receptor binding to the host cell or progeny virion release from the host cell. Anti-stem region antibodies hinder the membrane fusion process or induce antibody-dependent cytotoxicity to infected cells. In this study we identified a human monoclonal IgG1 antibody (CT302), which does not inhibit both the receptor binding and the membrane fusion process but efficiently reduced the nucleus entry of viral nucleoprotein suggesting a novel inhibition mechanism of viral infection by antibody. This antibody binds to the subtype-H3 hemagglutinin globular head domain of group-2 influenza viruses circulating throughout the population between 1997 and 2007.

No MeSH data available.


Related in: MedlinePlus

Reactivity of CT302 IgG1 to H3N2 trimeric HA.(A) After recombinant trimeric HA protein was coated onto 96-well plates, CT302 IgG1 (■) or negative-control anti-respiratory syncytial virus IgG1 (□) was incubated in wells as primary detection antibodies. Antibody bound to the recombinant His-tagged trimeric HA protein was detected with an HRP-conjugated anti-human IgG by addition of HRP substrate. Results represent the mean±S.D. obtained from duplicate wells per each condition. (B) Recombinant His-tagged trimeric HA proteins from H1, H3, and H5 strains were resolved by SDS-PAGE and transferred to nitrocellulose membranes, which were probed with CT302 IgG1 or anti-His antibody. (C) Amino acid sequences in the VH and VL regions of CT302 IgG1. HA, hemagglutinin; HRP, horseradish peroxidase.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4626144&req=5

pone.0141312.g001: Reactivity of CT302 IgG1 to H3N2 trimeric HA.(A) After recombinant trimeric HA protein was coated onto 96-well plates, CT302 IgG1 (■) or negative-control anti-respiratory syncytial virus IgG1 (□) was incubated in wells as primary detection antibodies. Antibody bound to the recombinant His-tagged trimeric HA protein was detected with an HRP-conjugated anti-human IgG by addition of HRP substrate. Results represent the mean±S.D. obtained from duplicate wells per each condition. (B) Recombinant His-tagged trimeric HA proteins from H1, H3, and H5 strains were resolved by SDS-PAGE and transferred to nitrocellulose membranes, which were probed with CT302 IgG1 or anti-His antibody. (C) Amino acid sequences in the VH and VL regions of CT302 IgG1. HA, hemagglutinin; HRP, horseradish peroxidase.

Mentions: Thirteen healthy volunteers who had not been vaccinated against influenza virus within the previous 2 years were vaccinated against the A/Uruguay/716/2007 (H3N2) strain. Almost all of the donor sera showed increased antibody titers after vaccination (data not shown). Using mRNA from mononuclear cells of the vaccinated group, we generated a scFv library with a complexity of 1.65×1010 clones. Next, we performed four rounds of biopanning and enzyme immunoassay to select phage bound to recombinant trimeric HA protein of the A/Brisbane/10/2007 strain, which shares the same amino acid sequence (except A138) with A/Uruguay/716/2007 (accession number CY121632.1). Twenty positive clones were identified among 192 colonies initially tested, and sequence analysis revealed 14 positive clones with unique amino-acid sequences. From these 14 positive clones, we selected the five most reactive clones to generate human anti-HA IgG1 antibodies. Reactivity of the antibodies to HA was confirmed by enzyme immunoassay and immunoblot analysis (Fig 1A and 1B, respectively).


An Anti-Influenza Virus Antibody Inhibits Viral Infection by Reducing Nucleus Entry of Influenza Nucleoprotein.

Yoon A, Yi KS, Chang SY, Kim SH, Song M, Choi JA, Bourgeois M, Hossain MJ, Chen LM, Donis RO, Kim H, Lee Y, Hwang do B, Min JY, Chang SJ, Chung J - PLoS ONE (2015)

Reactivity of CT302 IgG1 to H3N2 trimeric HA.(A) After recombinant trimeric HA protein was coated onto 96-well plates, CT302 IgG1 (■) or negative-control anti-respiratory syncytial virus IgG1 (□) was incubated in wells as primary detection antibodies. Antibody bound to the recombinant His-tagged trimeric HA protein was detected with an HRP-conjugated anti-human IgG by addition of HRP substrate. Results represent the mean±S.D. obtained from duplicate wells per each condition. (B) Recombinant His-tagged trimeric HA proteins from H1, H3, and H5 strains were resolved by SDS-PAGE and transferred to nitrocellulose membranes, which were probed with CT302 IgG1 or anti-His antibody. (C) Amino acid sequences in the VH and VL regions of CT302 IgG1. HA, hemagglutinin; HRP, horseradish peroxidase.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4626144&req=5

pone.0141312.g001: Reactivity of CT302 IgG1 to H3N2 trimeric HA.(A) After recombinant trimeric HA protein was coated onto 96-well plates, CT302 IgG1 (■) or negative-control anti-respiratory syncytial virus IgG1 (□) was incubated in wells as primary detection antibodies. Antibody bound to the recombinant His-tagged trimeric HA protein was detected with an HRP-conjugated anti-human IgG by addition of HRP substrate. Results represent the mean±S.D. obtained from duplicate wells per each condition. (B) Recombinant His-tagged trimeric HA proteins from H1, H3, and H5 strains were resolved by SDS-PAGE and transferred to nitrocellulose membranes, which were probed with CT302 IgG1 or anti-His antibody. (C) Amino acid sequences in the VH and VL regions of CT302 IgG1. HA, hemagglutinin; HRP, horseradish peroxidase.
Mentions: Thirteen healthy volunteers who had not been vaccinated against influenza virus within the previous 2 years were vaccinated against the A/Uruguay/716/2007 (H3N2) strain. Almost all of the donor sera showed increased antibody titers after vaccination (data not shown). Using mRNA from mononuclear cells of the vaccinated group, we generated a scFv library with a complexity of 1.65×1010 clones. Next, we performed four rounds of biopanning and enzyme immunoassay to select phage bound to recombinant trimeric HA protein of the A/Brisbane/10/2007 strain, which shares the same amino acid sequence (except A138) with A/Uruguay/716/2007 (accession number CY121632.1). Twenty positive clones were identified among 192 colonies initially tested, and sequence analysis revealed 14 positive clones with unique amino-acid sequences. From these 14 positive clones, we selected the five most reactive clones to generate human anti-HA IgG1 antibodies. Reactivity of the antibodies to HA was confirmed by enzyme immunoassay and immunoblot analysis (Fig 1A and 1B, respectively).

Bottom Line: To date, four main mechanisms mediating inhibition of influenza infection by anti-hemagglutinin antibodies have been reported.In this study we identified a human monoclonal IgG1 antibody (CT302), which does not inhibit both the receptor binding and the membrane fusion process but efficiently reduced the nucleus entry of viral nucleoprotein suggesting a novel inhibition mechanism of viral infection by antibody.This antibody binds to the subtype-H3 hemagglutinin globular head domain of group-2 influenza viruses circulating throughout the population between 1997 and 2007.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul National University, Seoul, South Korea; Cancer Research Institute, Seoul National University College of Medicine, Seoul National University, Seoul, South Korea.

ABSTRACT
To date, four main mechanisms mediating inhibition of influenza infection by anti-hemagglutinin antibodies have been reported. Anti-globular-head-domain antibodies block either influenza virus receptor binding to the host cell or progeny virion release from the host cell. Anti-stem region antibodies hinder the membrane fusion process or induce antibody-dependent cytotoxicity to infected cells. In this study we identified a human monoclonal IgG1 antibody (CT302), which does not inhibit both the receptor binding and the membrane fusion process but efficiently reduced the nucleus entry of viral nucleoprotein suggesting a novel inhibition mechanism of viral infection by antibody. This antibody binds to the subtype-H3 hemagglutinin globular head domain of group-2 influenza viruses circulating throughout the population between 1997 and 2007.

No MeSH data available.


Related in: MedlinePlus