Isolation and biological activities of an antineoplastic protein-polysaccharide complex (PS4A) obtained from Mycobacterium vaccae.

A mixture of water-soluble protein-polysaccharides (PS4A) was isolated by boiling intact cells of Mycobacterium vaccae, a fast growing mycobacterium. Sephadex G-75 column chromatography of the crude extract separated the biologically active high molecular weight (> 50 kDa) fraction (in the void volume) from the low molecular weight degradation products. Compositional analysis demonstrated that PS4A contained protein and polysaccharide in a ratio of approximately 1.5 to 1, but no lipids were detected. The antineoplastic activity was tested in vivo by a S-180 murine sarcoma model using female CFW mice. The immunostimulating activity was tested in vitro using murine peritoneal macrophages isolated from BALB/C mice. The results demonstrated that PS4A significantly decreased tumor incidence in vivo and produced activation of murine peritoneal macrophages. However, the antineoplastic activity was only attributable to the high molecular weight fraction of the protein-polysaccharide complex. The low molecular weight fraction had no antineoplastic activity in vivo despite stimulation of TNF-alpha production in vitro. In vitro experiments also demonstrated that although all PS4A components significantly increased TNF-alpha production by macrophages, the high molecular weight fraction stimulated more IL-1 production, indicating a better immunostimulating activity.