Stimulation of lymphocyte proliferation to monitor fractionation of thymus extracts.

Multi-step fractionations by solvent extractions, gel filtrations, ion-exchange chromatography, etc., of bovine thymus extracts were monitored by a standardized assay, in vitro, which led to the described peptides, thymones A, B and C. The assay uses spleen cells from neonatally thymectomized mice as tissue relevant to immunoregulation. Assay of incorporation of [3H]-thymidine into DNA located peaks of activities. Calf thymus outer fraction ("CTO") showed activity, but Fraction 5, synthetic thymosin alpha 1, [G1n1]-FTS, and glutathione, were inactive in this assay.