Viral Infection and Inactivation Protocols

For assays where cells were infected before being used to immunize mice, 293A cells (ATCC, CRL-1573) were used. For in vitro viral growth or plaque assays, BSC-1 cells (ATCC, CCL26) were used. For antigen presentation assays; MutuDCs (originally a gift from Hans Acha-Orbea, Lausanne, Switzerland) (56 (link)), 293A cells, and 293KbC2 cells (57 (link)) were used. Unmodified WR was originally a gift from B. Moss (National Institutes of Health, Bethesda, MD). HSV-1 strain KOS was provided by F. Carbone (University of Melbourne, Melbourne, Australia). Both were grown and titrated according to standard methods. Where indicated, virus or virus-infected cells were diluted in PBS and heat inactivated by incubation at 60°C for 1 h. Alternative inactivation methods included treatment of concentrated stocks with H₂O₂ as published (8 (link)) and with 1% paraformaldehyde for 20 min.