In this investigation, 34 structurally different flavonoids including derivatives of chalcone, flavanone, flavan-3-ol, flavone, flavonol, and their glycosides were evaluated for in vitro suppression of mitogen-induced lymphocyte proliferation and mixed lymphocyte culture from mouse spleen. Flavonoids, mainly derivatives of flavone and flavonol, clearly demonstrated the suppressive effects on lymphocyte proliferation at higher than 10(-6) M depending on the structures of flavonoid molecules, although their suppressive activities were less than that of cyclosporin A or prednisolone. Various glycosidic substitutions to A- and/or C-ring of the flavonoid aglycones were found to eliminate the suppressive activities of their aglycones, regardless of sugar compositions and positions of substitutions. In concanavalin A-induced lymphocyte proliferation, derivatives of flavone and flavonol having 2,3-unsaturation and at least 1 hydroxyl group showed the suppressive activity. In lipopolysaccharide-induced lymphocyte proliferation, only myricetin was active among flavonoids tested at the concentrations up to 10(-5) M. In mixed lymphocyte culture, some derivatives of flavone and flavonol with 2,3-unsaturation were active and especially flavone derivatives showed the higher suppressive activities than those of the flavonol derivatives.