【Abstract】 Objective To investigate the inflammatory response of Tolllike receptor in mediating severe handfootand mouth disease (HFMD). Methods Twentyseven children with severe HFMD hospitalized in Intensive Care Unit of Xuzhou Children's Hospital were enrolled in the study from July 2016 to January 2017. Thirty children with common HFMD were selected as disease control, and thirty healthy children as healthy control. Peripheral vein blood samples were collected from each individual, and used to isolate peripheral blood mononuclear cells (PBMCs) for mRNA extraction and sera for measuring levels of tumor necrosis factor α, interferon γ, interleukin 6. Results The expression levels of TLR3,TLR4, TLR7 and TLR8 mRNA in PBMCs of children with severe HFMD were significantly higher than those in the other children. The lowest values were found in the healthy control group (P<005). The expression levels of TLR2 and TLR9 mRNA were not significantly different among the groups(P＞005). Additionally, the expression levels of TNF α, IFN γ and IL 6 in the serum of the children in the severe group were significantly higher than those in the other two groups, and the lowest values were again found in the healthy control group (P<005). Pearson correlation analysis showed that the TLR3, TLR4, TLR7 and TLR8 mRNA levels in PBMCs were positively correlated with the TNF α, IFN γ, IL 6 levels in the serum (P<005). Conclusion The expression levels of TLR3, TLR4, TLR7 and TLR8 is higher, which then leads to the activation downstream signaling and affect the inflammatory response of the organism.