Isolation of conjunctival mucin and differential interaction with Pseudomonas aeruginosa strains of varied pathogenic potential
Isolation of conjunctival mucin and differential interaction with Pseudomonas aeruginosa strains of varied pathogenic potential
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DOI：
10.1016/j.exer.2003.08.007
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发表时间：
2003-12-01
影响因子：
3.4
通讯作者：
Willcox, MDP
中科院分区：
文献类型：
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作者：
Aristoteli, LP;Bojarski, B;Willcox, MDP
本研究的目的是调查不同致病潜力的铜绿假单胞菌菌株对纯化的眼粘蛋白的粘附。通过三个连续的密度梯度离心步骤纯化牛结膜粘蛋白。用从不同接触透镜事件中分离的生物素标记的细菌探测固定的粘蛋白，并通过光密度测定法定量。电镜下观察细菌皮利。结果表明，纯化的眼粘蛋白由含有至少一种杯状细胞来源的多分散高分子量群体组成，并与97 kDa粘蛋白相关蛋白相关。三种致病性铜绿假单胞菌菌株，Paer 1（57.5 +/- 10.8 x 10（6）CFU/ml（-1）;接触透镜诱导的急性红眼（克莱尔）），6294（127.0 +/- 4.7 x 10（6）CFU ml（-1）;微生物角膜炎）和Paer 25（60.5 +/- 11.3 x 10（6）CFU ml（-1）;克莱尔）显示出比阴性对照E显著更高的粘蛋白粘附水平。大肠杆菌（14.3 +/- 9.6 × 10（6）CFU ml（-1））（p < 0.005）。其余铜绿假单胞菌分离株Paer 3（无症状患者）、Paer 12（微生物角膜炎）和ATCC 15442（标准环境菌株）的粘蛋白粘附与阴性对照无显著差异。大多数测试的细菌菌株含有皮利;因此观察到的粘蛋白粘附差异不能仅由皮利状态解释。总之，铜绿假单胞菌分离株对纯化的眼粘蛋白表现出不同的粘附模式。有人提出，更多的贪婪粘蛋白粘附菌株有机会粘附，并随后穿透泪膜的粘液层，以启动发病机制。(C)2003 Elsevier Ltd.保留所有权利。
The purpose of the study was to investigate the adhesion of Pseudomonas aeruginosa strains with varying pathogenic potential to purified ocular mucin. Bovine conjunctival mucin was purified by three sequential density gradient centrifugation steps. Immobilised mucin was probed with biotin-labelled bacteria isolated from different contact lens events and quantified by densitometry. Bacterial pili were identified by electron microscopy. The results indicate that purified ocular mucin consisted of a polydisperse high molecular weight population containing at least one species of goblet cell origin and was associated with a 97 kDa mucin-associated protein. Three pathogenic P. aeruginosa strains, Paer1 (57.5 +/- 10.8 x 10(6) CFU ml(-1); contact lens induced acute red eye (CLARE)), 6294 (127.0 +/- 4.7 x 10(6) CFU ml(-1); microbial keratitis) and Paer25 (60.5 +/- 11.3 x 10(6) CFU ml(-1); CLARE) exhibited a significantly higher level of adhesion to mucin than the negative control, E. coli (14.3 +/- 9.6 x 10(6) CFU ml(-1)) (p < 0.005). The remaining P. aeruginosa isolates, Paer3 (asymptomatic patient), Paer12 (microbial keratitis) and ATCC15442 (standard environmental strain) did not significantly differ in their mucin adhesion from the negative control. The majority of bacterial strains tested contained pili; thus differences in mucin adhesion observed could not be solely explained by pili status. In conclusion, P. aeruginosa isolates exhibit differential adhesion patterns to purified ocular mucin. It is proposed that more avid mucin-adhering strains are given the opportunity to adhere and subsequently penetrate the mucous layer of the tear film to initiate pathogenesis. (C) 2003 Elsevier Ltd. All rights reserved.