Abstract
The LFBK-αvβ6 cell line is highly sensitive for the isolation of foot-and-mouth disease virus (FMDV) and porcinophilic vesicular viruses. However, LFBK-αvβ6 cells are contaminated with a non-cytopathic bovine viral diarrhea virus (BVDV), which complicates handling procedures in areas where other cell lines are maintained, as well downstream use of viral isolates. In this study, we used an aromatic cationic compound (DB772) to treat LFBK-αvβ6 cells using an approach that has been previously used to eliminate persistent BVDV from fetal fibroblast cell lines. After three cell passages with 4 μM DB772, BVDV could no longer be detected in unclarified cell suspensions using a pan-pestivirus real-time RT-PCR assay, and remained undetectable after treatment was stopped (nine passages) for an additional 28 passages. The analytical sensitivity of the DB772-treated LFBK-αvβ6 cultures (renamed WRL-LFBK-αvβ6) to titrations of FMDV and other vesicular virus isolates was comparable to untreated LFBK-αvβ6 cells. These new BVDV-free cells can be handled without the risk of cross-contaminating other cells lines or reagents, and used for routine diagnostics, in vivo studies and/or preparation of new vaccine strains.
Highlights
LFBK-αVβ6 cells are a porcine kidney cell line that was transduced to express the bovine αVβ6 integrin receptor in order to provide a highly sensitive, continuous cell line for the propagation of foot-and-mouth disease virus (FMDV) [(1); correction [2]]
Our results demonstrated that LFBK-αVβ6 cells had similar analytical sensitivity to FMDV epithelium suspensions as primary bovine thyroid cells (BTY), which had previously been identified as the most sensitive cell system for FMDV isolation [5]
WRL-LFBK-αvβ6 cell growth was slower during DB772 treatment relative to the untreated controls (10–40% based on estimates of daily confluency), but no cytotoxicity was observed
Summary
LFBK-αVβ6 cells are a porcine kidney cell line that was transduced to express the bovine αVβ6 integrin receptor in order to provide a highly sensitive, continuous cell line for the propagation of foot-and-mouth disease virus (FMDV) [(1); correction [2]]. LaRocco et al [1] demonstrated that over-expression of αVβ6 integrin, a known cellular receptor for FMDV, enhanced susceptibility to a range of FMDVs relative to other continuous cell lines (i.e., BHK, IB-RS-2, MVPK, LFBK, and LK), whilst maintaining sensitivity to vesicular viruses that are clinically indistinguishable from FMD. Fukai et al [3] demonstrated that LFBK-αVβ6 cells have similar susceptibility to non-epithelium FMDV clinical samples as ZZ-R 127 cells. The LFBK-αVβ6 cell line is persistently infected with a non-cytopathic bovine diarrhea virus (BVDV; family Flaviviridae, genus Pestivirus) (Rodriguez, personal communication), which complicates the use of these cells due to concerns about crosscontamination of other cell lines and downstream applications, including vaccine production and preparation of challenge viruses for in vivo studies
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