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.
Highlights
Human cytomegalovirus (HCMV) remains associated with considerable disease burden, despite the availability of antiviral medication [1]
The experiments showed that CAP cells supported the whole replication cycle of human cytomegalovirus (HCMV) and dense bodies (DBs)-release, rendering them exceptional to other E1A/E1B transformed cell lines
Is is characterized by by concomitant attempts of the HCMV infection infectionofofhuman humanfibroblasts fibroblasts characterized concomitant attempts of host cell to silence incoming viral genomes through the attachment of a repressive chromatin structure, the host cell to silence incoming viral genomes through the attachment of a repressive chromatin thereby shutting viral down gene expression
Summary
Human cytomegalovirus (HCMV) remains associated with considerable disease burden, despite the availability of antiviral medication [1]. A cell line was established from cells of fetal amniotic fluid by transformation with the early gene functions E1A and E1B of Ad5 [31] These cells (CAP; Cevec’s aminocyte production cell line; CEVEC Pharmaceuticals, Cologne, Germany) grow both anchor-dependent and in suspension and do not require animal serum for culture. Pilot experiments exposing CAP cells to HCMV in our laboratory provided surprising results, indicating a high level of initial viral infection and gene expression, leading to marked cytopathic alterations. This was opposed to what was known from other human cells transformed with. The experiments showed that CAP cells supported the whole replication cycle of HCMV and DB-release, rendering them exceptional to other E1A/E1B transformed cell lines
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