The Preliminary Study Of Inflammatory Effects Induced By Toll-like Receptor3and Toll-like Receptor7Activation In Respiratory Syncytial Virus Infected BV2Cells

Objective: To investigate the affect of respiratory syncytial virus (RSV) infectedmouse glioma cells (BV2) by activation of toll-like receptor3(TLR3) and toll-likereceptor7(TLR7), which induced the downstream inflammatory factors excessive inthe nervous system. In order to provide new idea for the prevention and clinical therapyof RSV infection.Method:(1) The infection of RSV in BV2cells were confirmed byimmunofluorescence using F proteins antibody at12,24, and36h post infection.(2)The quantitative of TLR3, TLR7mRNA were performed total RNA extracted by Trizolat4,8,12,16,24and36h post infection using RT-PCR. The uninfected cells were alsoharvested at same time-point using as control.(3) The expression levels of NF-kappa Bwere detected by Western blot at4,8,12,16,24and36h post infection. The uninfectedcells were also harvested at same time-point using as control.(4) The concentration ofTNF alpha in cell supernatant was detected by ELISA at4,8,12,16,24and36h postinfection. The uninfected cells were also harvested at same time-point using as control.Results: Results:(1) The green fluorescent signals were detected in RSV infected cellsand gradually increased from12h to36h post infection, but it was not lightened in thecontrol group.(2) The expression levels of TLR3, TLR7mRNA were increasesed inRSV infected cells detected by using RT-PCR, and there are significant differencesbetween infected cells and control at4,8,12,16,24and36h post infection. And theexaltation level was associated with the extension of infection. The expression levels of TLR3, TLR7mRNA were also significant differences at different time points in RSVinfected cells.(3) The expression levels of NF-kappa B in control group were lowerthan in RSV infected cells detected by western blots. There are significant differencesbetween infected cells and control at12,16,24and36h post infection (P<0.01). Theexpression levels of NF-kappa B were also significant differences at different timepoints in RSV infected cells.(4) The cell supernatant concentration of TNF alpha incontrol group was lower than in RSV infected cells detected by ELISA. There aresignificant differences between infected cells and control at12,16,24and36h postinfection (P<0.01). The cell supernatant concentration of TNF alpha was alsosignificant differences at different time points in RSV infected cells.Conclusion: RSV infection in BV2cells were induced inflammatory cytokines, such asTNF alpha secretion by activation of toll-like receptor3, toll-like receptor7. Theexcessive expression of inflammatory cytokines may be associated with Nervoussystem diseases.