Male genital tract inflammations constitute an important factor of male subfertility in which different mediators are discussed damaging sperm membranes. As acrosome reaction (AR) is ultimately a membrane function, this study aimed at investigating the effects of the chronic pelvic pain syndrome on AR. In a total of 56 patients with chronic pelvic pain syndrome NIH IIIA and NIH IIIB, AR, concentration of leucocytes in ejaculate and urine after prostate massage, sperm concentration, motility, vitality and morphology were determined. The control group (n = 95) consisted of patients with normozoospermia and proven absence of signs of inflammation. Compared to the control, both patients groups showed poorer sperm morphology (P < 0.0001) and sperm concentration (P < 0.002). In addition, while maximum AR was not affected, inducibility of AR was significantly lower (P < 0.004) and spontaneous AR increased (P < 0.02). Our data clearly demonstrate a significant influence of the inflammation on acrosomal functionality, which could be mediated by inflammatory mediators like ROS.