title: Porcine reproductive and respiratory syndrome virus-induced cell death exhibits features consistent with a nontypical form of apoptosis.
abstract: Porcine reproductive and respiratory syndrome virus (PRRSV), an arterivirus, belongs to a group of RNA viruses that are cytopathic for macrophages and establish persistent infections. Apoptosis is the presumed mechanism of cell death in monkey kidney cell lines and porcine alveolar macrophages after infection with European PRRSV isolates. However, evidence from in vivo and in vitro studies using North American strains have failed to identify apoptosis in cells supporting virus replication and suggest that apoptosis is present in only uninfected bystander cells. The purpose of this study was to evaluate the mechanism of cell death following the infection of MARC-145 cells with wild-type (P6) and a cell culture-adapted (P136) strains derived from the North American isolate SDSU-23983. At 2 days after infection with P136, cytoplasmic blebbing and nuclear condensation were absent in monolayers containing almost 90% infected cells. By day 3, these infected cells detached and showed evidence of apoptosis, including nuclear condensation and inter-nucleosomal DNA fragmentation. Apoptosis in single infected floating cells was confirmed by the co-localization of FITC-anti-digoxigenin antibody, used to detect uridine-digoxigenin-labled nuclear DNA in a terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling assay, and Texas red-labeled PRRSV antibody. A majority of infected floating cells were also positive for the uptake of trypan blue, an indicator of necrotic cell death. These results demonstrate that apoptosis does occur in PRRSV infected cells, but is a late event during PRRSV replication and rapidly culminates in a necrotic-like death.
authors.affiliation: Department of Biology-Microbiology, South Dakota State University, Brookings, SD 57007, USA.
authors.forename: Taeg Su
authors.initials: TS
authors.lastname: Kim
authors.forename: David A
authors.initials: DA
authors.lastname: Benfield
authors.forename: Raymond R R
authors.initials: RR
authors.lastname: Rowland
mesh_terms: D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002467:Cell Nucleus / Q000821:virology; D053938:DNA Fragmentation; D019316:Porcine respiratory and reproductive syndrome virus / Q000502:physiology*