Development of gene expression for FMDV-infected RAW264.7 cell

To determine whether type O FMDV-infected macrophage can release a variety of cytokines and activate innate immune responses, we have taken systematic functional genomics studies using microarray technology on differences expression gene of FMDV-infected and FMDV-uninfected mouse monocyte macrophage (RAW264.7 cells). We examined the expression patterns of 39,430 mouse genes probes, using Agilent mouse gene expression microarray. The results showed that 2541 genes were differentially expressed after FMDV infection, with 1207 up-regulated and 1334 down-regulated. Based on our previous data, the differential regulation of genes most associate with innate immune response. The mouse monocyte macrophage cell line RAW264.7 and BHK-21 cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum. O type FMDV was grown in BHK-21 cells for 8h at 37°C. The virus titers were detected using the Reed-Muench method to determine the 50% tissue culture infective dose. The supernatants of infected cells were clarified and stored at −80°C. RAW264.7 cells were infected or uninfected with O type FMDV at MOI of 2 for 1h at 37°C, and then removed the supernatant, washed with phosphate-buffered saline (PBS) (pH 7.4), and cultured with complete DMEM for 8h at 37°C.