Prostasomes are human-specific lipid vesicles originating from the prostate and present in seminal plasma. They are involved in a number of biological functions such as sperm motility and immunomodulation by seminal plasma. The aim of our study was to investigate whether prostasomes play a role in the antioxidant status of seminal plasma. In cell suspensions obtained after elimination of seminal plasma by centrifugation, reactive oxygen species (ROS) as measured by luminol-enhanced chemiluminescence were mainly produced by polymorphonuclear neutrophils (PMN). The addition of prostasomes to these cell suspensions lowered the overall ROS production in the basal state and after stimulation with phorbol ester. This action could not be explained by a ROS-scavenging capacity of the prostasomes, as demonstrated by their inability to scavenge ROS produced by 2,2'-azobis-2-amidinopropane dihydrochloride. Using electron spin resonance, we could assess the influence of prostasomes on the plasma membranes of blood PMN and show that it was characterized by an increase in the correlation-relaxation time of the probe 16-doxyl-stearic acid inserted in the membranes. Thus, prostasomes caused a rigidification of blood PMN membranes. These results strongly suggest that the effect of prostasomes in semen could result from their interaction with PMN.