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铜绿假单胞菌与正常及囊性纤维化气管支气管黏蛋白的差异性结合

Differential binding of Pseudomonas aeruginosa to normal and cystic fibrosis tracheobronchial mucins.

作者信息

Devaraj N, Sheykhnazari M, Warren W S, Bhavanandan V P

机构信息

Department of Biological Chemistry, M.S. Hershey Medical Center, Pennsylvania State University, Hershey 17033.

出版信息

Glycobiology. 1994 Jun;4(3):307-16. doi: 10.1093/glycob/4.3.307.

Abstract

Pseudomonas aeruginosa infection is a leading cause of deterioration of pulmonary function in patients with cystic fibrosis (CF). The interaction of the bacterium with CF and non-CF tracheobronchial mucins was examined to understand the biochemical basis for the high susceptibility of the lungs of CF patients to infection by P.aeruginosa. The binding of radiolabelled bacteria to pure mucins in solid-phase assays was not significantly above non-specific binding to various blocking agents, such as bovine serum albumin, Tween 20, milk powder and polyvinyl pyrrolidine. Further, there was a tendency for the bacteria to be excluded from plastic wells and membranes coated with mucin. Therefore, an indirect approach involving the binding of radiolabelled P.aeruginosa to asialo GM1 ganglioside, the putative receptor for the bacteria on tracheal cells, was used to compare the interaction of CF and non-CF mucins with the bacteria. Highly purified preparations of CF mucin were consistently better inhibitors of the binding of the bacteria to asialo GM1 ganglioside than non-CF mucin preparations. In the case of the binding of a stable mucoid strain, the difference was statistically significant (P < 0.001) at all concentrations of mucin tested. For the non-mucoid strain, the difference was significant only at the higher concentrations. Of the saccharides tested similarly, sialyl lactose and the oligosaccharide portion of asialo GM1 were found to be good inhibitors. The increased binding of the bacteria to CF mucin was further confirmed by a solution binding assay in which the binding of 125I-labelled mucin to unlabelled bacteria was determined.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

铜绿假单胞菌感染是囊性纤维化(CF)患者肺功能恶化的主要原因。研究了该细菌与CF和非CF气管支气管粘蛋白的相互作用,以了解CF患者肺部对铜绿假单胞菌感染高度易感的生化基础。在固相测定中,放射性标记细菌与纯粘蛋白的结合并不显著高于与各种封闭剂(如牛血清白蛋白、吐温20、奶粉和聚乙烯吡咯烷酮)的非特异性结合。此外,细菌有从涂有粘蛋白的塑料孔和膜上被排斥的趋势。因此,采用一种间接方法,即放射性标记的铜绿假单胞菌与气管细胞上该细菌的假定受体脱唾液酸GM1神经节苷脂结合,来比较CF和非CF粘蛋白与细菌的相互作用。高度纯化的CF粘蛋白制剂始终比非CF粘蛋白制剂更能有效抑制细菌与脱唾液酸GM1神经节苷脂的结合。对于稳定的粘液型菌株的结合,在所有测试的粘蛋白浓度下差异均具有统计学意义(P<0.001)。对于非粘液型菌株,差异仅在较高浓度时显著。在类似测试的糖类中,唾液酸乳糖和脱唾液酸GM1的寡糖部分被发现是良好的抑制剂。通过溶液结合试验进一步证实了细菌与CF粘蛋白结合的增加,该试验测定了125I标记的粘蛋白与未标记细菌的结合。(摘要截短于250字)

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