Toll-like receptors (TLRs) expressed on mast cells are essential for effective host defense against a wide variety of pathogens. Previous studies have demonstrated that TLR2 agonists Pam3CSK4 and PGN both stimulated IL-8 release in human mast cells. To determine the molecular basis for this phenomenon, we utilized a human mast cell line LAD2 cells. We found that only release of IL-8 stimulated by Pam3CSK4 was TLR2-mediated, which was confirmed by specific TLR2 shRNA.