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抗体介导的铜绿假单胞菌对上皮细胞黏附抑制作用的特性研究

Characterization of antibody-mediated inhibition of Pseudomonas aeruginosa adhesion to epithelial cells.

作者信息

Sexton M, Reen D J

机构信息

Children's Research Centre, Our Lady's Hospital for Sick Children, Dublin 12, Ireland.

出版信息

Infect Immun. 1992 Aug;60(8):3332-8. doi: 10.1128/iai.60.8.3332-3338.1992.

Abstract

An enzyme-linked immunosorbent assay system was developed and used to study adhesion of Pseudomonas aeruginosa to human epithelial cells and the abilities of specific antibodies to inhibit this process. Human buccal epithelial cells coated onto microtiter plates were incubated with P. aeruginosa suspensions, and adherent bacteria were detected by using anti-P. aeruginosa serum and a horseradish peroxidase-conjugated secondary antiserum. Adhesion, quantitated as an increase in A405, varied linearly with increasing numbers of bacterial CFU added per well in the range of 10(5) to 10(8) CFU per well. Adhesion of P. aeruginosa increased following trypsinization of buccal epithelial cells. Preincubation of bacteria with monoclonal antibodies directed against P. aeruginosa outer membrane protein H2 inhibited adhesion with all eight of the isolates tested. Preincubation of P. aeruginosa with sera from infected cystic fibrosis patients also resulted in inhibition of adhesion in the enzyme-linked immunosorbent assay system. This inhibitory activity was shown to be due to two factors: P. aeruginosa-specific immunoglobulin G and a non-immunoglobulin G serum component. These data support the hypothesis that bacterial components other than pili are involved in adhesion and suggest that anti-P. aeruginosa antibodies may be of use in preventing adhesion and subsequent colonization with P. aeruginosa.

摘要

开发了一种酶联免疫吸附测定系统,用于研究铜绿假单胞菌对人上皮细胞的黏附以及特异性抗体抑制该过程的能力。将包被在微量滴定板上的人颊上皮细胞与铜绿假单胞菌悬液孵育,使用抗铜绿假单胞菌血清和辣根过氧化物酶偶联的二抗血清检测黏附的细菌。黏附以A405的增加量进行定量,在每孔添加10(5)至10(8)CFU细菌的范围内,其与每孔添加的细菌CFU数量呈线性变化。颊上皮细胞经胰蛋白酶消化后,铜绿假单胞菌的黏附增加。用针对铜绿假单胞菌外膜蛋白H2的单克隆抗体对细菌进行预孵育,可抑制所测试的所有8株分离株的黏附。用囊性纤维化感染患者的血清对铜绿假单胞菌进行预孵育,在酶联免疫吸附测定系统中也导致黏附受到抑制。这种抑制活性被证明是由两个因素引起的:铜绿假单胞菌特异性免疫球蛋白G和一种非免疫球蛋白G血清成分。这些数据支持了菌毛以外的细菌成分参与黏附的假说,并表明抗铜绿假单胞菌抗体可能有助于预防铜绿假单胞菌的黏附和随后的定植。

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