Foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells

Foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells

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Conventional foot-and-mouth disease (FMD) vaccines are produced from virus grown in cell culture that is chemically inactivated by using binary ethylenimide (BEI). Here, we show that BEI treatment preserves both the architecture of FMDV particles, as inactivated viral particles showed by electron microscopy characteristics similar to those of infectious virions, as well as the general features of infectious virus internalization. Binding of inactivated particles to BHK-21 cells was blocked by preincubation with either a FMDV-specific monoclonal antibody or a synthetic peptide spanning the integrin-binding viral motif Arg-Gly-Asp (RGD). In addition, these particles were internalized into cultured cells through endocytosis, being directed to early endosomes, as indicated by their colocalization with the marker protein Rab5. When purified BEI-inactivated virions were labelled and their interaction with live cultured cells analyzed by time-lapse fluorescence microscopy, a major subpopulation of virus particles, about 80%, was shown to undergo internalization into a static endosome population, insensitive to the microtubule depolymerization exerted by nocodazole, while the remaining subpopulation (about 20%) was dynamic and sensitive to this drug. Thus, BEI-inactivated particles provide an interesting tool to study early steps in FMDV-cell interactions enabling a distinction between FMDV internalization and productive infection. Possible implications for FMDV immune response elicited following vaccine administration are discussed. © 2011 Elsevier Ltd.

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Main Authors: Martín-Acebes, M. A., Vázquez-Calvo, Á, González-Magaldi, M., Sobrino, F.
Format: journal article biblioteca
Language:eng
Published: 2011
Online Access:http://hdl.handle.net/20.500.12792/2679
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spelling dig-inia-es-20.500.12792-26792020-12-15T09:48:06Z Foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells Martín-Acebes, M. A. Vázquez-Calvo, Á, González-Magaldi, M. Sobrino, F. Conventional foot-and-mouth disease (FMD) vaccines are produced from virus grown in cell culture that is chemically inactivated by using binary ethylenimide (BEI). Here, we show that BEI treatment preserves both the architecture of FMDV particles, as inactivated viral particles showed by electron microscopy characteristics similar to those of infectious virions, as well as the general features of infectious virus internalization. Binding of inactivated particles to BHK-21 cells was blocked by preincubation with either a FMDV-specific monoclonal antibody or a synthetic peptide spanning the integrin-binding viral motif Arg-Gly-Asp (RGD). In addition, these particles were internalized into cultured cells through endocytosis, being directed to early endosomes, as indicated by their colocalization with the marker protein Rab5. When purified BEI-inactivated virions were labelled and their interaction with live cultured cells analyzed by time-lapse fluorescence microscopy, a major subpopulation of virus particles, about 80%, was shown to undergo internalization into a static endosome population, insensitive to the microtubule depolymerization exerted by nocodazole, while the remaining subpopulation (about 20%) was dynamic and sensitive to this drug. Thus, BEI-inactivated particles provide an interesting tool to study early steps in FMDV-cell interactions enabling a distinction between FMDV internalization and productive infection. Possible implications for FMDV immune response elicited following vaccine administration are discussed. © 2011 Elsevier Ltd. 2020-10-22T13:19:59Z 2020-10-22T13:19:59Z 2011 journal article http://hdl.handle.net/20.500.12792/2679 10.1016/j.vaccine.2011.10.031 eng Attribution-NonCommercial-ShareAlike 4.0 International http://creativecommons.org/licenses/by-nc-sa/4.0/ open access
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language eng
description Conventional foot-and-mouth disease (FMD) vaccines are produced from virus grown in cell culture that is chemically inactivated by using binary ethylenimide (BEI). Here, we show that BEI treatment preserves both the architecture of FMDV particles, as inactivated viral particles showed by electron microscopy characteristics similar to those of infectious virions, as well as the general features of infectious virus internalization. Binding of inactivated particles to BHK-21 cells was blocked by preincubation with either a FMDV-specific monoclonal antibody or a synthetic peptide spanning the integrin-binding viral motif Arg-Gly-Asp (RGD). In addition, these particles were internalized into cultured cells through endocytosis, being directed to early endosomes, as indicated by their colocalization with the marker protein Rab5. When purified BEI-inactivated virions were labelled and their interaction with live cultured cells analyzed by time-lapse fluorescence microscopy, a major subpopulation of virus particles, about 80%, was shown to undergo internalization into a static endosome population, insensitive to the microtubule depolymerization exerted by nocodazole, while the remaining subpopulation (about 20%) was dynamic and sensitive to this drug. Thus, BEI-inactivated particles provide an interesting tool to study early steps in FMDV-cell interactions enabling a distinction between FMDV internalization and productive infection. Possible implications for FMDV immune response elicited following vaccine administration are discussed. © 2011 Elsevier Ltd.
format journal article
author Martín-Acebes, M. A.
Vázquez-Calvo, Á, González-Magaldi, M.
Sobrino, F.
spellingShingle Martín-Acebes, M. A.
Vázquez-Calvo, Á, González-Magaldi, M.
Sobrino, F.
Foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells
author_facet Martín-Acebes, M. A.
Vázquez-Calvo, Á, González-Magaldi, M.
Sobrino, F.
author_sort Martín-Acebes, M. A.
title Foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells
title_short Foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells
title_full Foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells
title_fullStr Foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells
title_full_unstemmed Foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells
title_sort foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells
publishDate 2011
url http://hdl.handle.net/20.500.12792/2679
work_keys_str_mv AT martinacebesma footandmouthdiseasevirusparticlesinactivatedwithbinaryethylenimineareefficientlyinternalizedintoculturedcells
AT vazquezcalvoagonzalezmagaldim footandmouthdiseasevirusparticlesinactivatedwithbinaryethylenimineareefficientlyinternalizedintoculturedcells
AT sobrinof footandmouthdiseasevirusparticlesinactivatedwithbinaryethylenimineareefficientlyinternalizedintoculturedcells
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