In a continuing effort to investigate biological activities of flavonoids, nine biflavonoids, isolated from three plant sources were evaluated for their suppressive effects on mouse lymphocyte proliferation. The biflavonoids tested were amentoflavone, bilobetin, ginkgetin, isoginkgetin, sciadopitysin, ochnaflavone, 4'-O-methylochnaflavone, cryptomerin B and isocryptomerin. At 10 uM, several biflavonoids such as ginkgetin, isoginkgetin, ochnaflavone, cryptomerin B and isocryptomerin showed the suppressive activity against lymphocyte proliferation induced by Con A or LPS. Apigenin (flavone) and quercetin (flavonol) were suppressive against Con A-induced lymphocyte proliferation, but not against LPS-induced lymphocyte proliferation at the same concentration range. Biflavonoids were found to be irreversible inhibitors of lymphocyte proliferation. This is the first report describing the suppressive effects of naturally-occurring biflavonoids against lymphocyte proliferation.