Opisthorchis viverrini infection induces inflammation in and around the bile duct, leading to cholangiocarcinoma in humans. To examine the mechanism of O. viverrini-induced inflammatory response, we assessed the expression of Toll-like receptors (TLRs) in RAW 264.7 macrophage cell line treated with an extract of O. viverrini antigen. Flow cytometry and immunocytochemistry showed that O. viverrini antigen induced the expression of TLR2 but not TLR4. Western blotting and immunocytochemistry revealed that nuclear factor-kappaB (NF-kappaB), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were expressed in RAW 264.7 cells treated with O. viverrini antigen in a dose-dependent manner. These results suggest that O. viverrini induces inflammatory response through TLR2-mediated pathway leading to NF-kappaB-mediated expression of iNOS and COX-2.