Alginase treatment of mucoid Pseudomonas aeruginosa enhances phagocytosis by human monocyte-derived macrophages.
Mucoid Pseudomonas aeruginosa colonizes and infects the respiratory tract of most older patients with cystic fibrosis. These bacteria resist both opsonin-dependent and -independent phagocytosis by human polymorphonuclear leukocytes and monocyte-derived macrophages. Resistance to phagocytosis is thought to be mediated in part by the mucoid exopolysaccharide associated with the bacterial surface. The purpose of this study was to determine whether degradation of the mucoid exopolysaccharide by alginase enhances bacterial susceptibility to nonopsonic phagocytosis by macrophages. Eight phagocytosis-resistant mucoid P. aeruginosa isolates from patients with cystic fibrosis were studied. The bacteria were treated with a preparation of alginase from Bacillus circulans, and phagocytosis by macrophages was measured by a visual inspection assay. Alginase degradation of mucoid exopolysaccharide was measured by the periodate-thiobarbituric acid assay and by indirect immunofluorescence with a mouse monoclonal antibody to the mucoid exopolysaccharide. Alginase degraded the mucoid exopolysaccharide of all eight mucoid strains tested. Phagocytosis was enhanced in five of the eight strains. Alginase-enhanced phagocytosis was magnesium dependent and heat labile. Alginase may be a useful tool for studying the biological properties of P. aeruginosa mucoid exopolysaccharide.
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial / Q000032:analysis; D005455:Fluorescent Antibody Technique; D006026:Glycoside Hydrolases / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D008264:Macrophages / Q000502:physiology*; D008274:Magnesium / Q000494:pharmacology; D010587:Phagocytosis / Q000187:drug effects; D011133:Polysaccharide-Lyases; D011135:Polysaccharides, Bacterial / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000276:immunology / Q000472:pathogenicity*
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D011135:Polysaccharides, Bacterial; D006026:Glycoside Hydrolases; D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase; D008274:Magnesium