Isolation of Aleutian Disease Virus of Mink in Cell Culture

Abstract

Aleutian disease virus, the causative agent of a persistent infection in mink, was isolated in a continuous line of feline renal cells when the cultures were maintained at reduced temperature (31.8°). After serial in vitro passage of the virus at this temperature it had an optimum replication temperature of 37°. An immunofluorescence focus assay was found to be suitable for virus quantitation. The cultured virus reproduced Aleutian disease in mink, and the virus could be reisolated from the mink 10–180 days after inoculation. The properties of the virus suggest that it is a member of the parvovirus group.

A Novel Replicative Form DNA of Aleutian Disease Virus: the Covalently Closed Linear DNA of the Parvoviruses

“…It is possible that the higher degree of linkage observed in our system may reflect a lower activity of the postulated nickase in relation to the ADV RF DNA, perhaps resulting from the conditions required to propagate ADV in cell culture i.e. heterologous feline cells and incubation at 32 °C (Porter et al, 1977). These conditions may increase the ratio of ligated DNA to nicked DNA in the ADV RF DNA pool and allow even the detection of the Dccl RF structures.…”

The Capsid Proteins of Aleutian Mink Disease Virus Activate Caspases and Are Specifically Cleaved during Infection

“…AMDV has been successfully adapted to grow in CrFK cells (36,40,41). The initial isolate AMDV-Utah (40) is highly pathogenic to mink, causing persistent infection associated with severe dysfunction of the immune system (16,26).…”

“…For infection, CrFK cells were infected with AMDV-G at a multiplicity of infection (MOI) of 1 fluorescence focus-forming unit (FFU)/cell (16,40) and maintained at 31.8°C or 37°C for 6 or 7 days as specified.…”

“…Progeny virus was collected by a brief centrifugation of the cell lysates. Virus titers were determined by FFU in CrFK cells at 31.8°C as previously reported (16,40).…”

Persistent, Tolerant or Subacute Infection in Borna Disease Virus-infected Rats

“…Infectivity tests were performed using the second passage of primary newborn rabbit brain cells, which were grown in EDM, supplemented with 10 ~ inactivated foetal calf serum (Ludwig et al, 1973). The test is a fluorescent focus assay described by Porter et al (1977). Cells were seeded on round glass coverslips, which were placed in a 24-well plastic plate (Nunc) and incubated with 0.1 ml of 10-fold dilutions of virus suspension, followed by a medium change 1 day later.…”