Postpartum changes in the concentrations of IL-1 beta, IL-6, tumor necrosis factor-alpha(TNF-alpha), transforming growth factor-pi (TGF-beta 1), TGF-beta 2, and prostaglandin E-2 in 257 human milk samples collected longitudinally from 49 healthy mothers during the first 12 wk of lactation were determined by ELISA or RTA. The proinflammatory cytokines IL-1 beta, IL-6, and TNF-alpha were present in only a proportion of samples, and there was a wide range of concentrations detected at each time in the present study (IL-1 beta, <15-400 pg/mL; IL-6, <15-1032 pg/mL; TNF-alpha, (15-2933 pg/mL). Concentrations of prostaglandin E-2 increased after the first week and remained elevated for the remainder of the study (range, <10-9966 pg/mL). The antiinflammatory cytokines TGF-beta 1 (range, 43-7108 pg/mL) and TGF-beta 2 (range, 208-57935 pg/mL) were present in substantial quantities in all samples, and there was little change in the mean concentration during 12 wk of lactation. The present study shows that immunomodulating agents are normally present in human milk in physiologically relevant quantities for at least the first 3 mo of the breast-fed infant's life.