Recruitment of leukocytes to inflammatory sites is crucial in the pathogenesis of chronic inflammatory diseases. The aim of this study was to investigate if activation of CB₂ cannabinoid receptors would modulate the chemotactic response of human monocytes. Human monocytes treated with the CB₂ agonist JWH-015 for 12 to 18h showed significantly reduced migration to chemokines CCL2 and CCL3, associated with reduced mRNA and surface expression of their receptors CCR2 and CCR1. The induction of intercellular adhesion molecule (ICAM)-1 in response to interferon (IFN)- was inhibited by JWH-015. Moreover, JWH-015 cross-desensitized human monocytes for migration in response to CCL2 and CCL3 by its own chemoattractant properties. The CB₂ selective antagonist SR144528, but not the CB₁ antagonist SR147778 reversed the JWH-015-induced actions, while the CB₂ agonist JWH-133 mimicked the effects of JWH-015. The investigation of underlying pathways revealed involvement of phosphatidylinositol 3 kinase (PI3K)/Akt and extracellular signal-related kinase (ERK) 1/2, but not p38 mitogen activated protein kinase (MAPK). In conclusion, selective activation of CB₂ receptors modulates chemotaxis of human monocytes, which might have crucial effects in chronic inflammatory disorders such as atherosclerosis or rheumatoid arthritis.