Establishment of a BHK21 cell line persistently infected by the CATHAY strain of foot-and-mouth disease O virus
To investigate the molecular mechanism of the persistent infection of foot-and-mouth disease virus(FMDV),this research established a persistently infected cell line with O-type FMDV(CATHAY strain)and BHK21 cells using ammonium chloride,ribavirin,immune positive serum,and lower titer of FMDV.The persistent infection cell lines were screened and verified by RT-PCR,nucleic acid sequencing,immunofluorescence assay(IFA),and Western-blotting.The result showed that the BHK21 cell lines with FMDV persistent infection have been established by infection of lower titer of FMDV,the cell lines with FMDV can be continuously passaged.FMDV VP1 gene can be detectable from the 5th to 20th generation cells by RT-PCR and the sequences were highly homologous to the parental.The FMDV VP1 protein can be detectable by IFA in persistently infected cells.However,the remaining methods fail to develop the cell model of FMDV persistent infection due to their obvious toxic effects on cells.Therefore,we believed that the chemical drugs used in this study may be not a suitable reagent to develop the cell model of FMDV persistent infection,and direct infection using lower titer of FMDV is a simple and feasible method.Collectively,we successfully established BHK21 cell lines with FMDV persistent infection,which will helpful to investigate the interaction between FMDV and the host and clarifying the mechanism of FMDV persistent infection.
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