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Cell-to-cell transmission of HSV1 in human keratinocytes in the absence of the major entry receptor, nectin1

Herpes simplex virus 1 (HSV1) infects the stratified epithelia of the epidermis, oral or genital mucosa, where the main cell type is the keratinocyte. Here we have used nTERT human keratinocytes to generate a CRISPR-Cas9 knockout (KO) of the primary candidate HSV1 receptor, nectin1, resulting in a c...

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Autores principales: Kite, Joanne, Russell, Tiffany, Jones, Juliet, Elliott, Gillian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8505007/
https://www.ncbi.nlm.nih.gov/pubmed/34587223
http://dx.doi.org/10.1371/journal.ppat.1009631
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author Kite, Joanne
Russell, Tiffany
Jones, Juliet
Elliott, Gillian
author_facet Kite, Joanne
Russell, Tiffany
Jones, Juliet
Elliott, Gillian
author_sort Kite, Joanne
collection PubMed
description Herpes simplex virus 1 (HSV1) infects the stratified epithelia of the epidermis, oral or genital mucosa, where the main cell type is the keratinocyte. Here we have used nTERT human keratinocytes to generate a CRISPR-Cas9 knockout (KO) of the primary candidate HSV1 receptor, nectin1, resulting in a cell line that is refractory to HSV1 entry. Nonetheless, a small population of KO cells was able to support infection which was not blocked by a nectin1 antibody and hence was not a consequence of residual nectin1 expression. Strikingly at later times, the population of cells originally resistant to HSV1 infection had also become infected. Appearance of this later population was blocked by inhibition of virus genome replication, or infection with a ΔUL34 virus defective in capsid export to the cytoplasm. Moreover, newly formed GFP-tagged capsids were detected in cells surrounding the initial infected cell, suggesting that virus was spreading following replication in the original susceptible cells. Additional siRNA depletion of the second major HSV1 receptor HVEM, or PTP1B, a cellular factor shown elsewhere to be involved in cell-to-cell transmission, had no effect on virus spread in the absence of nectin1. Neutralizing human serum also failed to block virus transmission in nectin1 KO cells, which was dependent on the receptor binding protein glycoprotein D and the cell-to-cell spread glycoproteins gI and gE, indicating that virus was spreading by direct cell-to-cell transmission. In line with these results, both HSV1 and HSV2 formed plaques on nectin1 KO cells, albeit at a reduced titre, confirming that once the original cell population was infected, the virus could spread into all other cells in the monolayer. We conclude that although nectin1 is required for extracellular entry in to the majority of human keratinocytes, it is dispensable for direct cell-to-cell transmission.
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spelling pubmed-85050072021-10-12 Cell-to-cell transmission of HSV1 in human keratinocytes in the absence of the major entry receptor, nectin1 Kite, Joanne Russell, Tiffany Jones, Juliet Elliott, Gillian PLoS Pathog Research Article Herpes simplex virus 1 (HSV1) infects the stratified epithelia of the epidermis, oral or genital mucosa, where the main cell type is the keratinocyte. Here we have used nTERT human keratinocytes to generate a CRISPR-Cas9 knockout (KO) of the primary candidate HSV1 receptor, nectin1, resulting in a cell line that is refractory to HSV1 entry. Nonetheless, a small population of KO cells was able to support infection which was not blocked by a nectin1 antibody and hence was not a consequence of residual nectin1 expression. Strikingly at later times, the population of cells originally resistant to HSV1 infection had also become infected. Appearance of this later population was blocked by inhibition of virus genome replication, or infection with a ΔUL34 virus defective in capsid export to the cytoplasm. Moreover, newly formed GFP-tagged capsids were detected in cells surrounding the initial infected cell, suggesting that virus was spreading following replication in the original susceptible cells. Additional siRNA depletion of the second major HSV1 receptor HVEM, or PTP1B, a cellular factor shown elsewhere to be involved in cell-to-cell transmission, had no effect on virus spread in the absence of nectin1. Neutralizing human serum also failed to block virus transmission in nectin1 KO cells, which was dependent on the receptor binding protein glycoprotein D and the cell-to-cell spread glycoproteins gI and gE, indicating that virus was spreading by direct cell-to-cell transmission. In line with these results, both HSV1 and HSV2 formed plaques on nectin1 KO cells, albeit at a reduced titre, confirming that once the original cell population was infected, the virus could spread into all other cells in the monolayer. We conclude that although nectin1 is required for extracellular entry in to the majority of human keratinocytes, it is dispensable for direct cell-to-cell transmission. Public Library of Science 2021-09-29 /pmc/articles/PMC8505007/ /pubmed/34587223 http://dx.doi.org/10.1371/journal.ppat.1009631 Text en © 2021 Kite et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Kite, Joanne
Russell, Tiffany
Jones, Juliet
Elliott, Gillian
Cell-to-cell transmission of HSV1 in human keratinocytes in the absence of the major entry receptor, nectin1
title Cell-to-cell transmission of HSV1 in human keratinocytes in the absence of the major entry receptor, nectin1
title_full Cell-to-cell transmission of HSV1 in human keratinocytes in the absence of the major entry receptor, nectin1
title_fullStr Cell-to-cell transmission of HSV1 in human keratinocytes in the absence of the major entry receptor, nectin1
title_full_unstemmed Cell-to-cell transmission of HSV1 in human keratinocytes in the absence of the major entry receptor, nectin1
title_short Cell-to-cell transmission of HSV1 in human keratinocytes in the absence of the major entry receptor, nectin1
title_sort cell-to-cell transmission of hsv1 in human keratinocytes in the absence of the major entry receptor, nectin1
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8505007/
https://www.ncbi.nlm.nih.gov/pubmed/34587223
http://dx.doi.org/10.1371/journal.ppat.1009631
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