Eftekhar F, Speert D P
Department of Paediatrics, University of British Columbia, Vancouver, Canada.
Infect Immun. 1988 Nov;56(11):2788-93. doi: 10.1128/iai.56.11.2788-2793.1988.
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.
黏液型铜绿假单胞菌定殖并感染大多数老年囊性纤维化患者的呼吸道。这些细菌可抵抗人多形核白细胞和单核细胞衍生巨噬细胞的调理素依赖性及非依赖性吞噬作用。对吞噬作用的抗性被认为部分是由与细菌表面相关的黏液型胞外多糖介导的。本研究的目的是确定通过藻酸盐酶降解黏液型胞外多糖是否会增强细菌对巨噬细胞非调理吞噬作用的敏感性。研究了从囊性纤维化患者中分离出的8株抗吞噬作用的黏液型铜绿假单胞菌。用环状芽孢杆菌的藻酸盐酶制剂处理细菌,并通过目视检查法测定巨噬细胞的吞噬作用。通过高碘酸盐-硫代巴比妥酸测定法和用针对黏液型胞外多糖的小鼠单克隆抗体进行间接免疫荧光测定法来测量黏液型胞外多糖的藻酸盐酶降解情况。藻酸盐酶降解了所测试的所有8株黏液型菌株的黏液型胞外多糖。8株菌株中有5株的吞噬作用增强。藻酸盐酶增强的吞噬作用依赖于镁且对热不稳定。藻酸盐酶可能是研究铜绿假单胞菌黏液型胞外多糖生物学特性的有用工具。