Characterizing replication kinetics and plaque production of type I feline infectious peritonitis virus in three feline cell lines

对三种猫细胞系中I型猫传染性腹膜炎病毒的复制动力学和噬斑产生进行表征

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作者:Amornrat O'Brien ,Robert C Mettelman ,Aaron Volk ,Nicole M André ,Gary R Whittaker ,Susan C Baker

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. Keywords: AK-D cells; FIPV; Fcwf-4 cells; Feline coronavirus; Feline macrophage-like cell line; Plaque assay.

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