Activation of the Mitochondrial Apoptotic Signaling Platform during Rubella Virus Infection

Claus, Claudia; Manssen, Lena; Hübner, Denise; Roßmark, Sarah; Bothe, Viktoria; Petzold, Alice; Große, Claudia; Reins, Mareen; Mankertz, Annette; Frey, Teryl K.; Liebert, Uwe G.

Comparable to the vaccine strain HPV77, the selection of the Therien strain for its high replication rate in Vero cells was the result of a cell-culture adaptation process. CPE induction by a low-passaged clinical isolate, represented by the strain Wuerzburg-12 (Wb-12, RVi/Wuerzburg.DEU/47.11) was therefore addressed. The viral titer generated during Wb-12 infection is comparable to the one observed for Therien (representative graph in Figure 8A and [25] ). Additionally, CPE development by Wb-12 was comparable to Therien in terms of the onset of cell detachment. However, the number of floaters detected at 3 dpi per well of a six-well plate was 5 Figure 7 . Impact of RV infection on intracellular distribution of cyclophilin A. Immunofluorescence analysis was performed with anti-CypA antibody (shown in red) and anti-E1 antibody (shown in green) for RV-and mock-infected Vero cells (3 dpi). Nuclear DNA is shown in blue. Pink colour indicates overlap areas. As a control, overnight incubation with 0.003% hydrogen peroxide (H 2 O 2 ) was included. After fixation with 2% (w/v) paraformealdehyde, cells were permeabilized with 0.1% TritonX-100 (A) or with 0.3% Tween 20 (B). Arrows point to representative cells with a clear shift of CypA to the nucleus. Scale bar, 10 µm.