TLR2-mediated Immune Response Against HCV In Microglial BV-2 Cells

AIM To detected the expression of TLR2 and the levels of TNF-α,IFN-α,MCP-1,IL-6,IL-12 in supernatant of HCV-positive serum treated mice microglial cells BV-2, understanding immune response of microglia preliminary after stimulation of serum HCV-positive,Lay the foundation for the role of TLR2 in the pathogenesis of central nervous system HCV infected and its function in tissue injury.METHODS BV-2 cells were divided into the control group,the normal serum control group and the experimental group,cultrued with regular DMEM high glucose medium,200mL/L normal serum containing DMEM high glucose medium and 200mL/L HCV HCV-positive serum DMEM high glucose medium respectively.The expression of TLR2 mRNA was detected by RT-qPCR,and protein was detected by FCM,the level of TNF-α,IFN-α,MCP-1,IL-6 and IL-12 was measured with ELISA at 4 h,8 h,16 h ,24 h in each group after HCV infection.at the same time,we seted the TLR2 blocked group and it's control group,first,incubated 30 min with TLR2 blocked monoclonal antibody,then cultrued 24 h with 200mL/L HCV HCV-positive serum DMEM high glucose medium and 200mL/L normal serum containing DMEM high glucose medium respectively,measured the expression of TNF-α,IFN-α,MCP-1,IL-6 and IL-12 with ELISA in the supernatant of HCV infected mice microglial cells BV-2. All data were analyzed using SPSS13.0,one-way ANOVA used between groups,LSD were used among pairwise comparison,P<0.05 as significantly different.RESULTS The expression of TLR2 mRNA after infection appeared significantly increase,there was significant difference between control group and experiment group(P<0.05), there was no significant difference of the levels of TLR2 mRNA between the control group and the normal serum control group (P>0.05);BV-2 cells tagged with Mouse IgG1 K isotype control PE showed fluorescence peak;the expression of protein failed. the level of TNF-α,IFN-α,MCP-1,IL-6 and IL-12 in the experimental group were higher than in the control groups(P<0.05),which is decreased after TLR2 blocked,there was significant difference between 24 h(+) group and 24 h TLR2 blocked group(P<0.05)and there was no significant difference between the control of TLR2 blocked group and 24 h(-)(P>0.05).CONCLUSION TLR2 reside in BV-2 cells;Morphous of BV-2 cells altered after HCV-positive serum treatment,the expression of TLR2 mRNA and the level of TNF-α,IFN-α,MCP-1,IL-6 and IL-12 significantly increased in BV-2 cells after HCV-positive serum treatment, which might be involved in anti-HCV immunity.