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Mechanism of action and capsid-stabilizing properties of VHHs with an in vitro antipolioviral activity (doi: 10.1128/JVI.03402-13). / Schotte, Lise; Strauss, Mike; Thys, Bert; Halewyck, Hadewych; Filman, David J; Bostina, Mihnea; Hogle, James M.; Rombaut, Bartholomeus.

In: Journal of Virology, Vol. 88, 2014, p. 4403-4413.

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@article{e0ee6c3f6dd14c20aa95d19e5895b723,
title = "Mechanism of action and capsid-stabilizing properties of VHHs with an in vitro antipolioviral activity (doi: 10.1128/JVI.03402-13)",
abstract = "Previously, we reported on the in vitro antiviral activity of single-domain antibody fragments (VHHs) directed against poliovirus type 1. Five VHHs were found to neutralize poliovirus type 1 in an in vitro setting and showed 50{\%} effective concentrations in the nanomolar range. In the present study we further investigated the mechanism of action of these VHHs. All five VHHs interfere at multiple levels of the viral replication cycle, as they interfere both with attachment of the virus to cells and with viral uncoating. The latter effect is consistent with their ability to stabilize the poliovirus capsid, as observed ina thermo-fluor thermal shift assay, in which the virus is gradually heated and the temperature causing 50{\%} of the RNA to be released from the capsid is determined, either in the presence of in the absence of the VHHs. The VHH-capsid interactions were also seen to induce aggregation of the virus-VHH complexes. However, this observation cannot yet be linked to their mechanism of action. Cryo-electron microscopy (cryo-EM) reconstructions of two VHHs in complex with poliovirus type 1 show no conformational changes of the capsid to explain this aggregation. On the other hand, these reconstructions do show that the binding sites of VHHs PVSP6A and PVSP29F overlap the binding site for the poliovirus receptor (CD155/PVR) and span interfaces that are altered during receptor-induced conformational changes associated with cell entry. This may explain the interference at the level of cell attachment of the virus as well as their effect on uncoating.",
keywords = "poliovirus, capsid-stabilizing properties, VHHs, antipolioviral activity",
author = "Lise Schotte and Mike Strauss and Bert Thys and Hadewych Halewyck and Filman, {David J} and Mihnea Bostina and Hogle, {James M.} and Bartholomeus Rombaut",
year = "2014",
language = "English",
volume = "88",
pages = "4403--4413",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",

}

RIS

TY - JOUR

T1 - Mechanism of action and capsid-stabilizing properties of VHHs with an in vitro antipolioviral activity (doi: 10.1128/JVI.03402-13)

AU - Schotte, Lise

AU - Strauss, Mike

AU - Thys, Bert

AU - Halewyck, Hadewych

AU - Filman, David J

AU - Bostina, Mihnea

AU - Hogle, James M.

AU - Rombaut, Bartholomeus

PY - 2014

Y1 - 2014

N2 - Previously, we reported on the in vitro antiviral activity of single-domain antibody fragments (VHHs) directed against poliovirus type 1. Five VHHs were found to neutralize poliovirus type 1 in an in vitro setting and showed 50% effective concentrations in the nanomolar range. In the present study we further investigated the mechanism of action of these VHHs. All five VHHs interfere at multiple levels of the viral replication cycle, as they interfere both with attachment of the virus to cells and with viral uncoating. The latter effect is consistent with their ability to stabilize the poliovirus capsid, as observed ina thermo-fluor thermal shift assay, in which the virus is gradually heated and the temperature causing 50% of the RNA to be released from the capsid is determined, either in the presence of in the absence of the VHHs. The VHH-capsid interactions were also seen to induce aggregation of the virus-VHH complexes. However, this observation cannot yet be linked to their mechanism of action. Cryo-electron microscopy (cryo-EM) reconstructions of two VHHs in complex with poliovirus type 1 show no conformational changes of the capsid to explain this aggregation. On the other hand, these reconstructions do show that the binding sites of VHHs PVSP6A and PVSP29F overlap the binding site for the poliovirus receptor (CD155/PVR) and span interfaces that are altered during receptor-induced conformational changes associated with cell entry. This may explain the interference at the level of cell attachment of the virus as well as their effect on uncoating.

AB - Previously, we reported on the in vitro antiviral activity of single-domain antibody fragments (VHHs) directed against poliovirus type 1. Five VHHs were found to neutralize poliovirus type 1 in an in vitro setting and showed 50% effective concentrations in the nanomolar range. In the present study we further investigated the mechanism of action of these VHHs. All five VHHs interfere at multiple levels of the viral replication cycle, as they interfere both with attachment of the virus to cells and with viral uncoating. The latter effect is consistent with their ability to stabilize the poliovirus capsid, as observed ina thermo-fluor thermal shift assay, in which the virus is gradually heated and the temperature causing 50% of the RNA to be released from the capsid is determined, either in the presence of in the absence of the VHHs. The VHH-capsid interactions were also seen to induce aggregation of the virus-VHH complexes. However, this observation cannot yet be linked to their mechanism of action. Cryo-electron microscopy (cryo-EM) reconstructions of two VHHs in complex with poliovirus type 1 show no conformational changes of the capsid to explain this aggregation. On the other hand, these reconstructions do show that the binding sites of VHHs PVSP6A and PVSP29F overlap the binding site for the poliovirus receptor (CD155/PVR) and span interfaces that are altered during receptor-induced conformational changes associated with cell entry. This may explain the interference at the level of cell attachment of the virus as well as their effect on uncoating.

KW - poliovirus

KW - capsid-stabilizing properties

KW - VHHs

KW - antipolioviral activity

M3 - Article

VL - 88

SP - 4403

EP - 4413

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

ER -

ID: 2444267