This study investigates the ability of lymphocytes to utilize fatty acids originating from triacylglycerols and the effect of triacylglycerols upon mitogen-stimulated lymphocyte proliferation. Lymphocytes isolated from rat lymph nodes, spleen, thymus and lymphatic duct had a lipoprotein lipase activity of approx. 10 units/mg of protein, indicating that the fatty acids of circulating triacylglycerols are accessible to these cells. In culture, lymph node lymphocytes hydrolysed triacylglycerols added to the medium as emulsions. Both non-esterified fatty acids and free glycerol appeared in the cell culture medium, but their concentrations indicated that a high proportion of each (65-90% of fatty acids and 60-80% of glycerol) was taken up by the cells. The incorporation and fate of triacylglycerol-fatty acids was studied by culturing the cells in the presence of tri[H-3]oleoylglycerol or tri[C-14]linoleoylglycerol. Both fatty acids were incorporated into lymphocyte lipids in a time-dependent manner; linoleic acid was incorporated at a significantly greater rate than oleic acid. The majority of oleic acid (greater than 70%) was incorporated into cellular triacylglycerol, while less than 10% was incorporated into phospholipids. In contrast, linoleic acid incorporation into cellular triacylglycerol never exceeded 25%, while up to 45% was incorporated into phospholipids. Triacylglycerols containing polyunsaturated fatty acids inhibited concanavalin A-stimulated lymphocyte proliferation in a concentration- and time-dependent manner; triacylglycerols containing saturated fatty acids or oleic acid were not inhibitory. Such direct effects of certain triacylglycerols on lymphocyte function may explain why some clinical trials of polyunsaturated fatty acid-rich diets have been successful in improving the condition of patients suffering from inflammatory diseases.