Innate immune responses are rapid， dynamic and highly regulated to avoid overt reactions. This regulation is executed by innate immune tolerance mechanisms that remain obscure. Wnt5a is a signalling protein mainly involved in developmental processes and cancer. The effect of Wnt5a on inflammatory myeloid cells is controversial. Here， we combine primary cell cultures， in vitro binding studies， mass spectrometry and protein modelling to show that Wnt5a is a direct ligand of toll-like receptor (TLR) 2 and 4. The binding promotes a MyD88-non-canonical nuclear factor of kappa B (NFκB) and AP-1 signalling cascade， with contradictory profiles in mouse (pro-inflammatory) and human (anti-inflammatory) myeloid immune cells. These data reveal that the true nature of Wnt5a in inflammatory cells， is to regulate TLR signals， and in human myeloid cells it acts as an endogenous， tolerance-associated molecular pattern (TAMP)， inducing IL-10 and innate immune tolerance.