Yeast Mannans Inhibit Binding and Phagocytosis of Zymosan by Mouse Peritoneal Macrophages

We have examined the effects of various mannans, glycoproteins, oligosaccharides, monosaccharides, and sugar phosphates on the binding and phagocytosis of yeast cell walls (zymosan) by mouse peritoneal macrophages. A phosphonomannan ( PO4: mannose ratio = 1:8.6) from Kloeckera brevis was the most potent inhibitor tested; it inhibited binding and phagocytosis by 50% at concentrations of ∼3-5 μg/ml and 10 μg/ml, respectively. Removal of the phosphate from this mannan by mild acid and alkaline phosphatase treatment did not appreciably reduce its capacity to inhibit zymosan phagocytosis. The mannan from Saccharomyces cerevisiae mutant LB301 inhibits phagocytosis by 50% at 0.3 mg/ml, and a neutral exocellular glucomannan from Pichia pinus inhibited phagocytosis by 50% at 1 mg/ml. Cell wall mannans from wild type S. cerevisiae X2180, its mnn2 mutant which contains mannan with predominantly 1 → 6-linked mannose residues, yeast exocellular mannans and O-phosphonomannans were less efficient inhibitors requiring concentrations of 1-5 mg/ml to achieve 50% reduction in phagocytosis. Horseradish peroxidase, which contains high-mannose type oligosaccharides, was also inhibitory. Mannan is a specific inhibitor of zymosan binding and phagocytosis. The binding and ingestion of zymosan but not of IgG- or complement-coated erythrocytes can be obliterated by plating macrophages on substrates coated with poly-L-lysine (PLL)-mannan. Zymosan uptake was completely abolished by trypsin treatment of the macrophages and reduced by 50-60% in the presence of 10 mM EGTA. Pretreatment of the macrophages with chloroquine inhibited zymosan binding and ingestion. These results support the proposal that the macrophage mannose/ N-acetylglucosamine receptor (P. Stahl, J. S. Rodman, M. J. Miller, and P. H. Schlesinger, 1978, Proc. Natl. Acad. Sci. U. S. A. 75:1399-1403, mediates the phagocytosis of zymosan particles.