The purification of an immunosuppressive factor extracted from bovine spleen—III: Purification process

Abstract

A low mol. wt substance (PM < 2000) with immunosuppressive properties has been extracted from bovine spleen: when injected into mice, previously sensitized to sheep red blood cells, this factor reduced the hemolysin plaque forming capacity of spleen cells of treated animals measured vitro in the Jerne's test. Different techniques of purification: column chromatographies (Biogel P-2, neutral alumina, Sephadex LH-20), thin layer chromatography, high pressure liquid chromatography (μ Bondapack-C18), have been applied. They allowed the separation of the immunosuppressor from a stimulating factor which increased the response in the Jerne's test of the lymphocytes of the treated sensitized animals. Furthermore, those techniques led to a highly purified factor which is about 10 7 times more active than the starting material; the injection of a dose of approximately 2 × 10 −7 mg is sufficient to induce a 50% response inhibition in the Jerne's test.