This study analyzed the effects of a blocking anti-Toll-like receptor 2 (TLR2) antibody (T2.1) on TLR2 signaling and inflammatory cytokine production by bacterial lipoprotein (BLP) stimulated THP-1 cells. Cells were treated sequentially with anti-TLR2 antibody and BLP (A + B) or in the reverse order (B + A), or directly stimulated with BLP (B). Blank and isotype control groups were included. Tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) concentrations were measured by ELISA. Expression of nuclear factor κB (NF-κB), TLR2 and myeloid differentiation factor 88 (MyD88) was analyzed by Western Blot. Significantly higher expression of TNF-α, IL-6 and NF-κB was detected in all groups compared with the blank control (P < 0.05), although expression was significantly lower in A + B and B + A compared with the isotype control and B (P > 0.05). However, no significant differences were detected in TLR2 and MyD88 expression among these groups (P > 0.05). These data demonstrated that anti-TLR2 antibody significantly reduced TNF-α, IL-6 and NF-κB expression by BLP-stimulated THP-1 cells, thus indicating the protective value of this antibody in vivo.