Schaeffer L M, Weiss A A
Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati, Cincinnati, Ohio 45267, USA.
Infect Immun. 2001 Dec;69(12):7635-41. doi: 10.1128/IAI.69.12.7635-7641.2001.
The potential of human monocytes to mediate the clearance of Bordetella pertussis infection was examined. Bacteria expressing green fluorescent protein were incubated with adherent peripheral blood monocytes, and phagocytosis was quantified by using fluorescence microscopy. Monocytes internalized only a small percentage of the adherent bacteria. Surface-associated Bvg-regulated virulence factors, including adenylate cyclase toxin and filamentous hemagglutinin, did not affect attachment or phagocytosis. However, 1-h pretreatment with purified pertussis toxin inhibited the ability of monocytes to internalize wild-type bacteria. Mutations affecting the terminal trisaccharide of lipopolysaccharide resulted in reduced internalization without affecting adherence of bacteria to monocytes. Opsonization with human serum played only a modest role in promoting phagocytosis. The viability of internalized bacteria was determined by colony counts following treatment with polymyxin B and gentamicin. Less than 1% of internalized bacteria remained viable. These results suggest that pertussis toxin plays a role in the evasion of monocyte phagocytosis and that these cells represent a potential mediator of the clearance of B. pertussis infection.
研究了人类单核细胞介导清除百日咳博德特氏菌感染的潜力。将表达绿色荧光蛋白的细菌与贴壁外周血单核细胞一起孵育,并通过荧光显微镜对吞噬作用进行定量。单核细胞仅内化了一小部分贴壁细菌。表面相关的Bvg调节毒力因子,包括腺苷酸环化酶毒素和丝状血凝素,不影响细菌的附着或吞噬作用。然而,用纯化的百日咳毒素预处理1小时会抑制单核细胞内化野生型细菌的能力。影响脂多糖末端三糖的突变导致内化减少,但不影响细菌与单核细胞的黏附。用人血清进行调理在促进吞噬作用方面仅起适度作用。在用多粘菌素B和庆大霉素处理后,通过菌落计数确定内化细菌的活力。不到1%的内化细菌仍具有活力。这些结果表明,百日咳毒素在逃避单核细胞吞噬作用中起作用,并且这些细胞代表清除百日咳博德特氏菌感染的潜在介质。