| Autor: |
Krömmelbein N; Institute for Virology, University Medical Center of the Johannes Gutenberg-University Mainz, D-55131 Mainz, Germany. kroemme@uni-mainz.de., Wiebusch L; Department of Pediatric Molecular Biology, Charité University Medical Centre Berlin, D-10117 Berlin, Germany. lueder.wiebusch@charite.de., Schiedner G; CAP-CMV GmbH, D-51105 Cologne, Germany. schiedner@t-online.de., Büscher N; Institute for Virology, University Medical Center of the Johannes Gutenberg-University Mainz, D-55131 Mainz, Germany. bueschni@uni-mainz.de., Sauer C; Institute for Virology, University Medical Center of the Johannes Gutenberg-University Mainz, D-55131 Mainz, Germany. sauer.caroline@gmx.de., Florin L; Institute for Medical Microbiology and Hygiene, University Medical Center of the Johannes Gutenberg-University Mainz, D-55131 Mainz, Germany. lflorin@uni-mainz.de., Sehn E; Institute for Zoology, Johannes Gutenberg-University Mainz, D-55099 Mainz, Germany. sehn@uni-mainz.de., Wolfrum U; Institute for Zoology, Johannes Gutenberg-University Mainz, D-55099 Mainz, Germany. wolfrum@uni-mainz.de., Plachter B; 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. |
| Abstrakt: |
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. |
