The relationship between the occurrence of platelet-activating factor (PAF) and neutrophils in urine from patients with urinary tract infection was examined. PAF was detected in human pyuria, when leukocyte levels reached at least 300 cells/microL (n = 45), but not in normal urine (n = 12). The amount of PAF found in pyuria, measured by platelet aggregation assay, was 0.01 to 13.3 pmol/mL. A close correlation was seen between the amount of PAF present and the number of urinary leukocytes (p less than 0.01, r = 0.70). The leukocytes in pyuria consisted almost entirely of neutrophils (96 +/- 4%, mean +/- S.D.). Our findings suggest that the occurrence of PAF is associated with the accumulation of neutrophils in urine.