Antibody-mediated growth of influenza A NWS virus in macrophagelike cell line P388D1

Abstract

We investigated the internalization and growth of influenza A NWS virus in macrophagelike P388D1 cells. Flow cytometric analysis using fluorescein isothiocyanate-labeled virus showed that the attachment of normal rabbit serum-exposed virus (NS-V) to neuraminidase (NA)-treated cells was noticeably limited compared with that to untreated cells. However, rabbit antiserum-exposed virus (AS-V) could attach equally well to both cells. Virus coated with Fab prepared from antiviral immunoglobulin G could not attach. These data suggest that the NWS virus can infect P388D1 cells in one of two ways, via viral or via Fc receptors, depending on the presence of antibodies. The NS-V could grow in the untreated cells, but not in the NA-treated cells. The highest growth of AS-V in the NA-treated cells was observed at an antibody concentration showing 50% plaque reduction titer. Growth was exponentially decreased toward the lower and higher dilutions of antibodies. By using three different immunoglobulin G subclasses of monoclonal antibodies against hemagglutinin, it was demonstrated that both Fc receptors I and II could take part in this phenomenon. The presence of 20 mM NH4Cl inhibited the growth of both AS-V and NS-V, suggesting that the intracellular pathways after internalization via Fc or viral receptors are similar. These data indicate that the concentration of antibodies has a critical role on the antibody-mediated growth of influenza virus in macrophages.