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Viral gene expression and genome replication after HAdV-5 infection in primary cells and tumor cell lines.

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posted on 30.11.2011, 02:18 by Dominik E. Dorer, Frank Holtrup, Kurt Fellenberg, Johanna K. Kaufmann, Sarah Engelhardt, Jörg D. Hoheisel, Dirk M. Nettelbeck

Human primary cells (left panels) and tumor cell lines (right panels) were infected with HAdV-5 at titers resulting in 80% infection efficiency (see Fig. 1 for names of cell types; HBEC d1, HBEC d2 are HBEC from different donors). Inoculums were removed after one hour incubation. Total RNA and DNA was harvested for every indicated time point and was analyzed for E1A (A) and fiber (C) mRNA levels and for viral genome copies (B), respectively, by qPCR. Results of representative experiments are shown; repetition experiments yielded identical time points for the onset of E1A and fiber mRNA expression and virus genome replication. For HBEC d2, genome copy numbers were not determined at 20 h and 24 h, because of limited numbers of cells from this donor.