A common feature underlying active states of inflammation is the migration of neutrophils (PMNs) from the circulation and across a number of tissue barriers in response to chemoattractant stimuli. Although our group has recently established a discreet role for the PMN chemoattractant, hepoxilin A₃ (HXA₃) in the process of PMN recruitment, very little is known regarding the interaction of HXA₃ with PMNs. To characterize further the event of HXA₃-induced PMN transepithelial migration, we sought to determine the adhesion molecules required for migration across different epithelial surfaces (T84 intestinal and A549 airway cells) relative to two well-studied PMN chemoattractants, formyl-methionyl-leucyl-phenylalanine (fMLP) and leukotriene B₄ (LTB₄). Our findings reveal that the adhesion interaction profile of PMN transepithelial migration in response to HXA₃ differs from the adhesion interaction profile exhibited by the structurally related eicosanoid LTB₄. Furthermore, unique to PMN transepithelial migration induced by gradients of HXA₃ was the critical dependency of all four major surface adhesion molecules examined (i.e. CD18, CD47, CD44 and CD55). Our results suggest that the particular chemoattractant gradient imposed, as well as the type of epithelial cell monolayer, each plays a role in determining the adhesion molecules involved in transepithelial migration. Given the complexities of these interactions, our findings are important to consider with respect to adhesion molecules that may be targeted for potential drug development.