Replication for herpes virus made more efficient
Members of the European project ANTICANCER RETINOID aimed to investigate the action of retinoid compounds against cancer. Past research has demonstrated that retinoids display very definite anti-cancer action favouring apoptosis or simply stopping tumourigenesis. Project objectives also included work on retinoid anti-viral action. As part of the viral research, Italian-based project partners at the University of Ferrara developed a method to prepare cell-free inoculum using the Human herpes virus 8 (HHV-8). Human herpes virus 8 causes Kaposi's sarcoma which results in tumours on the skin and digestive and respiratory tracts. The initial step was to obtain the purified and concentrated virus from activated lymphoma cell lines followed by high-speed centrifugation. The next procedure was to find a cell line which would enable the virus to replicate at high efficiency. Vero cells were used, their source being kidney epithelial cells. HHV-8 naturally targets primary endothelial cells. However, the team found that viral production was even more efficient in the Vero cell lines than in primary endothelial cells. The incorporation of promoters of HHV-8 replication to speed up the process was the next phase. The trials used the open reading frame 30 (ORF30) sequence in particular as an established promoter for the virus. The efficient replication of viruses like HHV-8 represents a crucial part for understanding the repressive action of retinoids at different stages of the viral life cycle. The further elucidation of the biochemical processes involved may be used in the continuing fight against cancer.
