Development and characterization of stable cell lines constitutively expressing the porcine reproductive and respiratory syndrome virus nucleocapsid protein

Abstract

Despite global efforts to control porcine reproductive and respiratory syndrome virus (PRRSV) infection, the virus continues to cause economic problems in the swine industry worldwide. In this study, we attempted to generate and characterize a panel of stable BHK cell lines that constitutively express the nucleocapsid (N) protein of type 1 or type 2 PRRSV. The established BHK cell lines were found to react well with N-specific antibodies as well as the hyperimmune serum of pigs raised against each genotype of PRRSV. Taken together, the data implicate a potential usefulness for the newly generated stable cell lines as a diagnostic reagent for PRRSV serology.

Pathogenicity and genetic characteristics associated with cell adaptation of a virulent porcine reproductive and respiratory syndrome virus nsp2 DEL strain CA-2

“…Briefly, MARC-145 cells were seeded in 96-well cell culture plates and infected with CA-2-12 or CA-2-100 for 24 h at 37 C in a 5% CO 2 incubator. The virus-infected cells were fixed and subsequently subjected to IPMA using serial 2-fold dilutions of the sera (starting dilution 1:4) as described previously (Decorte et al, 2014;Sagong et al, 2010). The results were determined by examination with a light microscope (Leica, Wetzlar, Germany), and the IPMA titer was calculated as the highest dilution of serum giving PRRSV-specific cytoplasmic staining of infected MARC-145 cells.…”

Development of a novel immunoperoxidase monolayer assay for detection of swine Hepatitis E virus antibodies based on stable cell lines expressing the ORF3 protein

“…Customarily, an IPMA is used to monitor antigen by inoculating viruses into cells (Khudyakov Yu et al, 1994;Direksin et al, 2002;Guedes et al, 2002), which involves complicated procedures and unstable antigen content. A recent study suggested that cells stably expressing the major viral antigen could be used for IPMA (Sagong et al, 2010). In the present study, we developed an IPMA for the detection of swHEV antibodies using modified HepG2 cell lines that stably expressed the ORF3 protein as a diagnostic target.…”

Cultivation and comparison of BHK-21 anchorage semi-dependent cell line in different production systems

“…Radlett et al [11] greatly improved the growth conditions of BHK-21 cells in suspension culture demonstrating the usefulness of BHK-21 cells for the commercial production of viral vaccines for animals. After that, cell lines have been used for virus detection and production of virus and recombinant proteins in many laboratories throughout the world [12][13][14][15][16]. Cultivation of the BHK cells in submerged systems similar to microorganisms may well be considered an advance for industrial use of animal cells.…”