Characterizing replication kinetics and plaque production of type I feline infectious peritonitis virus in three feline cell lines
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Rescue and Characterization of Recombinant Virus from a New World Zika Virus Infectious Clone
Published on: June 7, 2017
Amornrat O'Brien1, Robert C Mettelman1, Aaron Volk1
1Department of Microbiology and Immunology, Loyola University of Chicago, Stritch School of Medicine, Maywood, IL, United States.
Abstract:
Investigating type I feline coronaviruses (FCoVs) in tissue culture is critical for understanding the basic virology, pathogenesis, and virus-host interactome of these important veterinary pathogens. This has been a perennial challenge as type I FCoV strains do not easily adapt to cell culture. Here we characterize replication kinetics and plaque formation of a model type I strain FIPV Black in Fcwf-4 cells established at Cornell University (Fcwf-4 CU). We determined that maximum virus titers (>107 pfu/mL) were recoverable from infected Fcwf-4 CU cell-free supernatant at 20 h post-infection. Type I FIPV Black and both biotypes of type II FCoV formed uniform and enumerable plaques on Fcwf-4 CU cells. Therefore, these cells were employable in a standardized plaque assay. Finally, we determined that the Fcwf-4 CU cells were morphologically distinct from feline bone marrow-derived macrophages and were less sensitive to exogenous type I interferon than were Fcwf-4 cells purchased from ATCC.
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