Identification of a new cell line permissive to porcine reproductive and respiratory syndrome virus infection and replication which is phenotypically distinct from MARC-145 cell line

Abstract

BackgroundAirborne transmitted pathogens, such as porcine reproductive and respiratory syndrome virus (PRRSV), need to interact with host cells of the respiratory tract in order to be able to enter and disseminate in the host organism. Pulmonary alveolar macrophages (PAM) and MA104 derived monkey kidney MARC-145 cells are known to be permissive to PRRSV infection and replication and are the most studied cells in the literature. More recently, new cell lines developed to study PRRSV have been genetically modified to make them permissive to the virus. The SJPL cell line origin was initially reported to be epithelial cells of the respiratory tract of swine. Thus, the goal of this study was to determine if SJPL cells could support PRRSV infection and replication in vitro.ResultsThe SJPL cell growth was significantly slower than MARC-145 cell growth. The SJPL cells were found to express the CD151 protein but not the CD163 and neither the sialoadhesin PRRSV receptors. During the course of the present study, the SJPL cells have been reported to be of monkey origin. Nevertheless, SJPL cells were found to be permissive to PRRSV infection and replication even if the development of the cytopathic effect was delayed compared to PRRSV-infected MARC-145 cells. Following PRRSV replication, the amount of infectious viral particles produced in SJPL and MARC-145 infected cells was similar. The SJPL cells allowed the replication of several PRRSV North American strains and were almost efficient as MARC-145 cells for virus isolation. Interestingly, PRRSV is 8 to 16 times more sensitive to IFNα antiviral effect in SJPL cell in comparison to that in MARC-145 cells. PRRSV induced an increase in IFNβ mRNA and no up regulation of IFNα mRNA in both infected cell types. In addition, PRRSV induced an up regulation of IFNγ and TNF-α mRNAs only in infected MARC-145 cells.ConclusionsIn conclusion, the SJPL cells are permissive to PRRSV. In addition, they are phenotypically different from MARC-145 cells and are an additional tool that could be used to study PRRSV pathogenesis mechanisms in vitro.

Replacement of Porcine CD163 Scavenger Receptor Cysteine-Rich Domain 5 with a CD163-Like Homolog Confers Resistance of Pigs to Genotype 1 but Not Genotype 2 Porcine Reproductive and Respiratory Syndrome Virus

“…For example, Shanmukhappa et al (20) showed that nonpermissive BHK cells transfected with a CD151 plasmid acquired the ability to support PRRSV replication, and incubation with a polyclonal anti-CD151 antibody reduced the infection of MARC-145 cells. In addition, a simian cell line, SJPL, originally developed for use in propagating swine influenza viruses, was shown by Provost et al (21) to support PRRSV replication. Important properties of the SJPL cell line include the presence of CD151 and the absence of CD163.…”

The non-structural protein Nsp2TF of porcine reproductive and respiratory syndrome virus down-regulates the expression of Swine Leukocyte Antigen class I

“…This cell line was cultured in Minimum Essential Medium (MEM) supplemented with 0.2% hygromycin, 10% FBS, non-essential amino acid (NEAA, Invitrogen, GibcoBRL) and sodium pyruvate (Invitrogen, GibcoBRL), and maintained in MEM with 5% FBS after virus infection. Primary porcine alveolar macrophages (PAMs) were prepared from lung lavage fluids of PRRSV-negative piglets as previously described (Provost et al, 2012), and were maintained in RPMI 1640 medium (Invitrogen) supplemented with 10% FBS. Monocyte-derived DCs (MoDCs) were prepared as described previously (Subramaniam et al, 2011).…”

Generation and characterization of a porcine endometrial endothelial cell line susceptible to porcine reproductive and respiratory syndrome virus

“…In addition, different batches of PAM are not always equally susceptible to PRRSV, and it is necessary to test each batch before use. On the other hand, although the non-porcine permissive immortalized cell line MARC-145 is less sensitive cell type, it is routinely used for in vitro propagation of PRRSV (including viral titration) and for large scale production of PRRSV vaccine strains (Provost et al, 2012). Our viral titration assay showed that the viral titer of PEE-grown PRRSV was slightly lower or higher than that of PAM-or MARC-145-grown virus (Fig.…”