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Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication

The human cytomegalovirus (HCMV) replicates to high titers in primary human fibroblast cell cultures. A variety of primary human cells and some tumor-derived cell lines do also support permissive HCMV replication, yet at low levels. Cell lines established by transfection of the transforming function...

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Autores principales: Krömmelbein, Natascha, Wiebusch, Lüder, Schiedner, Gudrun, Büscher, Nicole, Sauer, Caroline, Florin, Luise, Sehn, Elisabeth, Wolfrum, Uwe, Plachter, Bodo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4776192/
https://www.ncbi.nlm.nih.gov/pubmed/26848680
http://dx.doi.org/10.3390/v8020037
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author Krömmelbein, Natascha
Wiebusch, Lüder
Schiedner, Gudrun
Büscher, Nicole
Sauer, Caroline
Florin, Luise
Sehn, Elisabeth
Wolfrum, Uwe
Plachter, Bodo
author_facet Krömmelbein, Natascha
Wiebusch, Lüder
Schiedner, Gudrun
Büscher, Nicole
Sauer, Caroline
Florin, Luise
Sehn, Elisabeth
Wolfrum, Uwe
Plachter, Bodo
author_sort Krömmelbein, Natascha
collection PubMed
description The human cytomegalovirus (HCMV) replicates to high titers in primary human fibroblast cell cultures. A variety of primary human cells and some tumor-derived cell lines do also support permissive HCMV replication, yet at low levels. Cell lines established by transfection of the transforming functions of adenoviruses have been notoriously resistant to HCMV replication and progeny production. Here, we provide first-time evidence that a permanent cell line immortalized by adenovirus type 5 E1A and E1B (CAP) is supporting the full HCMV replication cycle and is releasing infectious progeny. The CAP cell line had previously been established from amniotic fluid cells which were likely derived from membranes of the developing fetus. These cells can be grown under serum-free conditions. HCMV efficiently penetrated CAP cells, expressed its immediate-early proteins and dispersed restrictive PML-bodies. Viral DNA replication was initiated and viral progeny became detectable by electron microscopy in CAP cells. Furthermore, infectious virus was released from CAP cells, yet to lower levels compared to fibroblasts. Subviral dense bodies were also secreted from CAP cells. The results show that E1A/E1B expression in transformed cells is not generally repressive to HCMV replication and that CAP cells may be a good substrate for dense body based vaccine production.
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spelling pubmed-47761922016-03-09 Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication Krömmelbein, Natascha Wiebusch, Lüder Schiedner, Gudrun Büscher, Nicole Sauer, Caroline Florin, Luise Sehn, Elisabeth Wolfrum, Uwe Plachter, Bodo Viruses Article The human cytomegalovirus (HCMV) replicates to high titers in primary human fibroblast cell cultures. A variety of primary human cells and some tumor-derived cell lines do also support permissive HCMV replication, yet at low levels. Cell lines established by transfection of the transforming functions of adenoviruses have been notoriously resistant to HCMV replication and progeny production. Here, we provide first-time evidence that a permanent cell line immortalized by adenovirus type 5 E1A and E1B (CAP) is supporting the full HCMV replication cycle and is releasing infectious progeny. The CAP cell line had previously been established from amniotic fluid cells which were likely derived from membranes of the developing fetus. These cells can be grown under serum-free conditions. HCMV efficiently penetrated CAP cells, expressed its immediate-early proteins and dispersed restrictive PML-bodies. Viral DNA replication was initiated and viral progeny became detectable by electron microscopy in CAP cells. Furthermore, infectious virus was released from CAP cells, yet to lower levels compared to fibroblasts. Subviral dense bodies were also secreted from CAP cells. The results show that E1A/E1B expression in transformed cells is not generally repressive to HCMV replication and that CAP cells may be a good substrate for dense body based vaccine production. MDPI 2016-02-02 /pmc/articles/PMC4776192/ /pubmed/26848680 http://dx.doi.org/10.3390/v8020037 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Krömmelbein, Natascha
Wiebusch, Lüder
Schiedner, Gudrun
Büscher, Nicole
Sauer, Caroline
Florin, Luise
Sehn, Elisabeth
Wolfrum, Uwe
Plachter, Bodo
Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication
title Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication
title_full Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication
title_fullStr Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication
title_full_unstemmed Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication
title_short Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication
title_sort adenovirus e1a/e1b transformed amniotic fluid cells support human cytomegalovirus replication
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4776192/
https://www.ncbi.nlm.nih.gov/pubmed/26848680
http://dx.doi.org/10.3390/v8020037
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