The effects of dietary supplementation with ω‐3‐polyunsaturated fatty acids (ω‐3‐PUFA) on the proliferative response of PBMC and on the secretion of monokines and arachidonic acid metabolites from PBMC and monocytes (Mo) from healthy subjects and patients with recent‐onset insulin‐dependent diabetes mellitus (IDDM) were examined. Three groups of eight to nine healthy individuals were randomized to either 2.0 g/day or 4.0 g/day of ω‐3‐PUFA devoid of vitamins A and D, or an isocaloric amount of placebo. Furthermore, eight patients with recent‐onset IDDM received 4.0 g/day of ω‐3‐PUFA. IL‐lβ production and TNF‐α secretion was determined before and after 7 weeks of treatment, and 10 weeks after withdrawal of treatment. Significant increases in platelet and PBMC membrane eicosapentaenoic acid was found in ω‐3‐PUFA‐treated individuals. ω‐3‐PUFA treatment significantly reduced the content of IL‐Ib in lysates of PBMC, but did not affect PBMC or Mo secretion of IL‐1β, TNF‐α or prostaglandin E2 (PGE2) or PBMC leukotriene B4 (LTB4) secretion in healthy subjects or in IDDM patients. A significant inhibition of the PHA‐stimulated. but not the spontaneous or PPD‐stimulated, proliferative response of PBMC was observed in healthy and diabetic subjects treated with (o‐3‐PUFA. No correlation was found between PHA‐stimulated PBMC proliferation and PBMC secretion of TNF‐α and IL‐1β. There were no significant differences in the spontaneous or the PPD‐or PHA‐stimulated proliferative responses of PBMC between diabetic and healthy individuals at entry. We conclude that although dietary supplementation with 4.0 g/day of ω‐3‐PUFA inhibits the proliferation of PBMC and reduces IL‐I immunoreactivity in PBMC and Mo, it does not alter monokine, PGE2 or LTB4, secretion in healthy or IDDM subjects.