SIMULTANEOUS IDENTIFICATION OF VIRAL-PROTEINS AND NUCLEIC-ACIDS IN CELLS INFECTED WITH ALEUTIAN MINK DISEASE PARVOVIRUS

A method combining in situ hybridization and immunohistochemistry was used to characterize cells infected with Aleutian mink disease parvovirus (ADV). Single-stranded RNA hybridization probes specific for obligate replicative intermediates and antisera specific for virion or nonstructural proteins were employed. Crandell feline kidney cells in which the ADV-G strain of ADV was permissively replicating contained virion and non-structural proteins, large amounts of single stranded virion DNA, duplex replicative form (RF) DNA, and mRNA. Late in the infectious cycle, however, cells containing non-structural proteins but little nucleic acid were observed, probably representing cells in the end stage of viral cytopathology. Sections of lung prepared from mink kits infected with the ADV-Utah 1 strain were then examined. Alveolar type II cells permissively replicating ADV contained viral nucleic acids and proteins in patterns nearly identical to CRFK cells, suggesting that permissive ADV replication was similar in vitro and in vivo. Another population of ADV containing cells that had cytoplasmic virion antigen, but undetectable levels of non-structural protein was found in vivo. Furthermore, although virion DNA was present in the cytoplasm of these cells, RF DNA or mRNA could not be detected. These cells may have been alveolar macrophages sequestering viral particles. © 1990.