Abstract
BackgroundFoot-and-mouth disease virus (FMDV) is able to cause persistent infection in ruminants besides acute infection and disease. Since the mechanisms of viral persistence and the determining factors are still unknown, in vitro systems help explore and reveal mechanisms of persistence in vivo by providing useful models for the study of RNA genome mutations and evolution. Ammonium chloride, a lysosomotropic agent that raises intralysosomal pH, reduces the yield of FMDV during infection of BHK-21 cells.ResultsThe persistent infection with FMDV serotype O in BHK-21 cells was selected and established readily after treatment of ammonium chloride that acts primarily on the cells. Intact virions were observed located inside the endosomes. Viral genome RNAs and specific proteins were detected in the persistent cells to validate the establishment of viral persistence. Infection of the persistent viruses could not form plaques in host cells but virulence was enhanced. Basing on analysis and comparison of cDNA sequences of resident viruses and wild type viruses, 15 amine acid mutations were found, all of which were located in nonstructural proteins rather than in structural proteins.ConclusionsTherefore, persistent infection of cell cultures with FMDV is successfully established with some distinctive features. It would be worthwhile to further investigate the mechanisms of viral persistence.
Highlights
Foot-and-mouth disease virus (FMDV) is able to cause persistent infection in ruminants besides acute infection and disease
Detection of viral genome RNAs in acutely infected BHK21 cells maintained in the presence and absence of ammonium chloride To investigate the effect of ammonium chloride on the virus yield post-infection, BHK-21 cells in 12-well plates were infected with FMDV and maintained in 5% FBS MEM supplemented with different concentrations of ammonium chloride
The results demonstrated that ammonium chloride can reduce the yield of FMDV during infection of BHK-21 cells
Summary
Foot-and-mouth disease virus (FMDV) is able to cause persistent infection in ruminants besides acute infection and disease. Foot-and-mouth disease virus (FMDV), the aetiological agent of foot-and-mouth disease (FMD), which belongs to the Aphthovirus genus of the Picornaviridae family [6], usually causes an acute, systematic infection of clovenhooved animals and often produces a persistent noncytocidal infection of ruminants [7]. The FMDV genome consists of a single plus-sense stranded RNA of approximately 8.5 kb in length, which contains a single open reading frame flanked by two non-coding regions and a small viral protein VPg linked covalently to the 5’end [9,10]. Replication of FMDV genome RNA occurs via a complementary negative strand RNA.
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