Investigating Macrophage Immune Response to MHV68 Infection

Wild-type MHV68 viral stocks were prepared and titers determined on NIH 3T12 cells. Bone marrow-derived macrophages or MEFs were infected with live or UV-inactivated MHV68 at the indicated multiplicity of infection (MOI) for 1 h to allow adsorption and were washed 2 to 3 times with phosphate-buffered saline (PBS) prior to medium replenishment. UV inactivation of MHV68 was performed as previously described (17 (link)). Supplementation experiments were performed by adding the indicated compound(s) to the replenishment media following viral adsorption. GW3965 and TOFA [tetradecyloxy-2-furoic acid] were purchased from Cayman Chemical (Ann Arbor, MI). GW3965 was dissolved in dimethyl sulfoxide (DMSO) and then diluted in cell culture media to a concentration of 5 µM. TOFA was dissolved in DMSO and then diluted in cell culture media to a concentration of 10 µg/ml. Control cell cultures were treated with equivalent amounts of DMSO.