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Marek's disease virus infection of phagocytes: a de novo in vitro infection model

Marek’s disease virus (MDV) is an alphaherpesvirus that induces T-cell lymphomas in chickens. Natural infections in vivo are caused by the inhalation of infected poultry house dust and it is presumed that MDV infection is initiated in the macrophages from where the infection is passed to B cells and...

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Autores principales: Chakraborty, Pankaj, Vervelde, Lonneke, Dalziel, Robert G., Wasson, Peter S., Nair, Venugopal, Dutia, Bernadette M., Kaiser, Pete
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Microbiology Society 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5656796/
https://www.ncbi.nlm.nih.gov/pubmed/28548038
http://dx.doi.org/10.1099/jgv.0.000763
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author Chakraborty, Pankaj
Vervelde, Lonneke
Dalziel, Robert G.
Wasson, Peter S.
Nair, Venugopal
Dutia, Bernadette M.
Kaiser, Pete
author_facet Chakraborty, Pankaj
Vervelde, Lonneke
Dalziel, Robert G.
Wasson, Peter S.
Nair, Venugopal
Dutia, Bernadette M.
Kaiser, Pete
author_sort Chakraborty, Pankaj
collection PubMed
description Marek’s disease virus (MDV) is an alphaherpesvirus that induces T-cell lymphomas in chickens. Natural infections in vivo are caused by the inhalation of infected poultry house dust and it is presumed that MDV infection is initiated in the macrophages from where the infection is passed to B cells and activated T cells. Virus can be detected in B and T cells and macrophages in vivo, and both B and T cells can be infected in vitro. However, attempts to infect macrophages in vitro have not been successful. The aim of this study was to develop a model for infecting phagocytes [macrophages and dendritic cells (DCs)] with MDV in vitro and to characterize the infected cells. Chicken bone marrow cells were cultured with chicken CSF-1 or chicken IL-4 and chicken CSF-2 for 4 days to produce macrophages and DCs, respectively, and then co-cultured with FACS-sorted chicken embryo fibroblasts (CEFs) infected with recombinant MDV expressing EGFP. Infected phagocytes were identified and sorted by FACS using EGFP expression and phagocyte-specific mAbs. Detection of MDV-specific transcripts of ICP4 (immediate early), pp38 (early), gB (late) and Meq by RT-PCR provided evidence for MDV replication in the infected phagocytes. Time-lapse confocal microscopy was also used to demonstrate MDV spread in these cells. Subsequent co-culture of infected macrophages with CEFs suggests that productive virus infection may occur in these cell types. This is the first report of in vitro infection of phagocytic cells by MDV.
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spelling pubmed-56567962017-11-14 Marek's disease virus infection of phagocytes: a de novo in vitro infection model Chakraborty, Pankaj Vervelde, Lonneke Dalziel, Robert G. Wasson, Peter S. Nair, Venugopal Dutia, Bernadette M. Kaiser, Pete J Gen Virol Research Article Marek’s disease virus (MDV) is an alphaherpesvirus that induces T-cell lymphomas in chickens. Natural infections in vivo are caused by the inhalation of infected poultry house dust and it is presumed that MDV infection is initiated in the macrophages from where the infection is passed to B cells and activated T cells. Virus can be detected in B and T cells and macrophages in vivo, and both B and T cells can be infected in vitro. However, attempts to infect macrophages in vitro have not been successful. The aim of this study was to develop a model for infecting phagocytes [macrophages and dendritic cells (DCs)] with MDV in vitro and to characterize the infected cells. Chicken bone marrow cells were cultured with chicken CSF-1 or chicken IL-4 and chicken CSF-2 for 4 days to produce macrophages and DCs, respectively, and then co-cultured with FACS-sorted chicken embryo fibroblasts (CEFs) infected with recombinant MDV expressing EGFP. Infected phagocytes were identified and sorted by FACS using EGFP expression and phagocyte-specific mAbs. Detection of MDV-specific transcripts of ICP4 (immediate early), pp38 (early), gB (late) and Meq by RT-PCR provided evidence for MDV replication in the infected phagocytes. Time-lapse confocal microscopy was also used to demonstrate MDV spread in these cells. Subsequent co-culture of infected macrophages with CEFs suggests that productive virus infection may occur in these cell types. This is the first report of in vitro infection of phagocytic cells by MDV. Microbiology Society 2017-05 2017-05-26 /pmc/articles/PMC5656796/ /pubmed/28548038 http://dx.doi.org/10.1099/jgv.0.000763 Text en © 2017 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the terms of the Creative Commons Attribution 4.0 International License (http://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
Chakraborty, Pankaj
Vervelde, Lonneke
Dalziel, Robert G.
Wasson, Peter S.
Nair, Venugopal
Dutia, Bernadette M.
Kaiser, Pete
Marek's disease virus infection of phagocytes: a de novo in vitro infection model
title Marek's disease virus infection of phagocytes: a de novo in vitro infection model
title_full Marek's disease virus infection of phagocytes: a de novo in vitro infection model
title_fullStr Marek's disease virus infection of phagocytes: a de novo in vitro infection model
title_full_unstemmed Marek's disease virus infection of phagocytes: a de novo in vitro infection model
title_short Marek's disease virus infection of phagocytes: a de novo in vitro infection model
title_sort marek's disease virus infection of phagocytes: a de novo in vitro infection model
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5656796/
https://www.ncbi.nlm.nih.gov/pubmed/28548038
http://dx.doi.org/10.1099/jgv.0.000763
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