title: Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication.
abstract: 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.
authors: Institute for Virology, University Medical Center of the Johannes Gutenberg-University Mainz, D-55131 Mainz, Germany. kroemme@uni-mainz.de.
authors: Natascha
authors: N
authors: Krömmelbein
authors: Department of Pediatric Molecular Biology, Charité University Medical Centre Berlin, D-10117 Berlin, Germany. lueder.wiebusch@charite.de.
authors: Lüder
authors: L
authors: Wiebusch
authors: CAP-CMV GmbH, D-51105 Cologne, Germany. schiedner@t-online.de.
authors: Gudrun
authors: G
authors: Schiedner
authors: Institute for Virology, University Medical Center of the Johannes Gutenberg-University Mainz, D-55131 Mainz, Germany. bueschni@uni-mainz.de.
authors: Nicole
authors: N
authors: Büscher
authors: Institute for Virology, University Medical Center of the Johannes Gutenberg-University Mainz, D-55131 Mainz, Germany. sauer.caroline@gmx.de.
authors: Caroline
authors: C
authors: Sauer
authors: Institute for Medical Microbiology and Hygiene, University Medical Center of the Johannes Gutenberg-University Mainz, D-55131 Mainz, Germany. lflorin@uni-mainz.de.
authors: Luise
authors: L
authors: Florin
authors: Institute for Zoology, Johannes Gutenberg-University Mainz, D-55099 Mainz, Germany. sehn@uni-mainz.de.
authors: Elisabeth
authors: E
authors: Sehn
authors: Institute for Zoology, Johannes Gutenberg-University Mainz, D-55099 Mainz, Germany. wolfrum@uni-mainz.de.
authors: Uwe
authors: U
authors: Wolfrum
authors: Institute for Virology, University Medical Center of the Johannes Gutenberg-University Mainz, D-55131 Mainz, Germany. plachter@uni-mainz.de.|Research Center for Immunotherapy, University Medical Center of the Johannes Gutenberg-University Mainz, D-55131 Mainz, Germany. plachter@uni-mainz.de.
authors: Bodo
authors: B
authors: Plachter
mesh_terms: D017500:Adenovirus E1A Proteins / Q000235:genetics / Q000378:metabolism*; D017501:Adenovirus E1B Proteins / Q000235:genetics / Q000378:metabolism*; D000258:Adenovirus Infections, Human / Q000821:virology*; D000260:Adenoviruses, Human / Q000235:genetics / Q000378:metabolism*; D002472:Cell Transformation, Viral; D003587:Cytomegalovirus / Q000235:genetics / Q000502:physiology*; D003586:Cytomegalovirus Infections / Q000821:virology*; D006801:Humans; D014779:Virus Replication
chemical_list: D017500:Adenovirus E1A Proteins; D017501:Adenovirus E1B Proteins
keywords: CAP; Cevec’s aminocyte production cell line; adenovirus E1A/E1B; amniotic fluid cells; cytomegalovirus