Establishment And Identification Of Cell Models Persistently Infected With Foot And Mouth Disease Virus & Its Microcalorimetric Study

Baby hamster kidney passage 21 clone 13(BHK-21 c-13, kidney, Syrian or golden hamster Mesocricetus auratus, from ATCC and perserved by CCTCC) were infected respectively by foot and mouth disease virus serum type O (FMDV, from Lanzhou Veterinary Research Institute CAAS). After selection and persistent subculture, steady models of cells persistently infected with FMDV were established, hence O-BHK-RN cell lines were obtained. In order to identify the basic viral features in O-BHK-RN cells, RT-PCR assay, transmission electron microscopy observation and flow cytometry analysis were used in the experiment. The results showed all passages of infected host cells still harbored FMDV and they had the radical characteristics of viral persistent infection. The existence of FMDV did not influence cell cycle of host cells obviously. And the rate of Go/G1, S, G2/M cells to total cells almost remained the same compare with normal BHK-21 cell. It proved that the cell models of persistently infected cell line O-BHK-RN were established.In the meantime both microcalorimetric measurement analysis by using LKB2277 bioactivity monitor and traditional method used in one-step growth curve in virology were carried out on BHK-21 cell line infected by FMDV. The microcalorimetric FMDV infection curve is consistent with traditional method used in one-step growth curve in virology. In succession, we checked the BHK-21 cell microcalorimetric curves of temperature ,, pharmaceutical effect(interferon). The BHK-21 cell microcalorimetric curves of temperature,pharmaceutical effect(interferon) were sensitive to the change of the cell culture environment.Conclusively, the establishment of the two steady models of host cells persistently infected with FMDV and the parallel/vertical research on these cell models should be the foundation for the further research of FMDV persistent infection. The results of microcalorimetric FMDV infection analysis show that microcalorimetric study can be applied in virus infection research through measuring metabolic energy released from BHK-21 cell and BHK-21 cell infected by FMDV. Furthermore this microcalorimetric method provided a kind of new dynamic continuous analysis method on the virus infection research.