Enhancement of infectivity of murine cytomegalovirus in vitro by centrifugal inoculation

Abstract

Centrifugation of murine cytomegalovirus inocula from a variety of sources onto secondary mouse embryo cell monolayers at 1,900 x g for 30 min regularly revealed 10- to 100-fold more infectious virus than could be found in the same materials using standard inoculation methods. Virus demonstrable only by centrifugation was present throughout the entire growth cycle in a constant proportion to virus measured without centrifugation. Extracellular growth curves of both populations revealed an 18- to 21-hr latent period, followed by a long-linear increase over the next 12 hr; final yield was 30 plaque-forming units (PFU) per cell. Centrifugation of cells prior to inoculation or after standard adsorption and removal of inoculum failed to result in any significant change in measured virus titer. However, even after 4-hr adsorption, the supernatant inoculum could be transferred and centrifuged onto a fresh monolayer resulting in the same increment of measurable virus. Neutralizing antibody and interferon were equally efficacious against 100 PFU of virus as defined by either method. Thus, this newly identified population of cytomegalo-virus represents the vast majority of potentially infectious units and appears to differ solely in ease of adsorption onto cell monolayers.