We examined the surface expression of lactoferrin by human neutrophils. Western blot analysis with anti-lactoferrin antibodies demonstrated the presence of a 78- to 79-kDa band in plasma membranes isolated from resting neutrophils that corresponded to the 78-to 79-kDa protein in neutrophil secondary granules. Flow cytometry using FITC-conjugated anti-lactoferrin antibodies confirmed that lactoferrin is expressed on the neutrophil surface. Preincubating the neutrophils in acidic (pH 3.9) buffer did not alter staining of the cells by the antibodies. Surface expression of lactoferrin was also detected on neutrophils in whole blood. Neutrophil activation by C5a or the calcium ionophore A23187 did not increase the surface expression of lactoferrin. Instead, the level of lactoferrin expression detected with one of two monoclonal antibodies was diminished after neutrophil activation, suggesting a possible conformational change in the lactoferrin. The surface-expressed lactoferrin may provide a mechanism for the interaction between lactoferrin-binding microorganisms and neutrophils.