Selection and characterization of canine, swine and rabbit cell lines resistant to bovine viral diarrhea virus

Contamination of cell cultures with adventitious viruses may pose serious risks for virology diagnosis, research and vaccine production. This article reports the selection and characterization of three cell lines resistant to bovine viral diarrhea virus (BVDV), a major contaminant of cell cultures. The resistant cells were obtained from canine (MDCK), swine (PK-15) and rabbit (RK-13) parental cell lines by expanding and cloning cells that survived lytic infection with BVDV. All three selected cell lines were resistant to three standard BVDV strains and to 10 field isolates as demonstrated by immunofluorescence for viral antigens and by co-cultivation with susceptible cells. Inoculation of selected cells with BVDV (10 TCID50/cell) resulted in frequencies of infection of <10−5 (MDCK-R, PK-15R) and 3.3 × 10−4 (RK-13R). Comparing to the parental cell lines, these cells were >10,000-, >20,000- and 600-fold less susceptible to BVDV, respectively. Inoculation of resistant cells with BVDV in the presence of polyethylene–glycol increased the frequency of infection in the order of >437-, >346- and 87-fold, respectively, indicating that resistance is probably due to a block in viral entry. Nevertheless, each resistant cell line retained the susceptibility to three other viruses that replicate in the parental cells. Thus, these cells may be useful for many virology purposes, with a very low risk of BVDV contamination.