Isolation of virus-free Herpesvirus saimiri antigen-positive plasma membrane vesicles

Abstract

Experiments were undertaken to determine whether virus-free, antigen-positive plasma membrane vesicles could be isolated from owl monkey kidney cells infected with Herpesvirus saimiri. The results demonstrate that vesicles can be produced by using a vesiculation fluid containing 25 mM formaldehyde/2 mM dithiothreitol. Electron microscopy revealed that these vesicles were free of detectable virus particles. Vesicles prepared from the infected cells contained virus-induced membrane antigens as shown by membrane immunofluorescence and by inhibition of antibody-dependent lymphocyte cytotoxicity. Nonhuman primates immunized with vesicles produced antibodies to these membrane antigens, late cytoplasmic antigens, and neutralizing antibodies. Infectious virus was not demonstrated in these vesicles by cocultivation with owl monkey kidney cells or by the inoculation of cottontop marmosets. Furthermore, no DNA could be demonstrated in vesicles prepared from Herpesvirus-infected owl monkey kidney cells. The implication of these findings in relation to the question of a virus-free membrane vaccine against Herpesvirus infections is discussed.