Adherence status regulates the primary cellular activation responses to the flavivirus West Nile

Increases in cell-surface intercellular adhesion molecule-1 (ICAM-1; CD54) and major histocompatibility complex antigen class I (MHC-I) and class II (MHC-II) expression during flavivirus infection of murine macrophages was strongly dependent on adherence status. CD54 and MHC expression was significantly increased during infection with the flavivirus West Nile (WNV) on adherent, but not on non-adherent, macrophages. In contrast, increased CD54 and MHC-I expression was induced by interferon-gamma (IFN-gamma) in both cultures but was significantly greater on adherent cells than non-adherent cells. Adherent status was also important in human embryonic fibroblasts (HEF), adherent cells of non-immune origin. Similar to macrophages, WNV induced increased CD54 or MHC-I expression on adherent but not non-adherent HEF. Again, induction of these antigens by IFN-gamma occurred in both cultures but was significantly greater on adherent cells than non-adherent cells. Macrophages or HEF that aggregated when cultured at high density under non-adherent conditions, responded to WNV and IFN-gamma in a manner similar to adherent cells. Unresponsive non-adherent cells infected with WNV or treated with IFN-gamma under non-adherent conditions for 24 or 48 hr recovered if transferred to adherent culture conditions for 24 or 48 hr. Moreover, these cells expressed significantly higher cell-surface CD54 and MHC-I concentrations, compared to similarly treated HEF cultured under adherent conditions during the entire culture period. WNV infection also induced significant nitric oxide production in macrophages, and adherence status was similarly important in this response, with adherent cells producing higher amounts of nitrite/nitrate than non-adherent cells. These results suggest that adherent status may be critical for effective antiviral immune responses involving macrophages.