Fluorescent labels influence phagocytosis of Bordetella pertussis by human neutrophils.
Fluorescent labels influence phagocytosis of Bordetella pertussis by human neutrophils.
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荧光标记影响人中性粒细胞对百日咳博德特氏菌的吞噬作用。
DOI：
10.1128/iai.67.8.4264-4267.1999
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发表时间：
1999
影响因子：
3.1
通讯作者：
Weiss,AA
中科院分区：
文献类型：
--
作者：
Weingart,CL;Broitman-Maduro,G;Dean,G;Newman,S;Peppler,M;Weiss,AA
为探讨中性粒细胞吞噬功能在宿主防御百日咳杆菌中的作用，用异硫氰酸荧光素（FITC）或绿色荧光蛋白（GFP）标记百日咳杆菌，并与粘附的人中性粒细胞在热灭活的人免疫血清存在或不存在的情况下孵育。在不存在抗体的情况下，FITC标记的细菌主要位于中性粒细胞的表面上，几乎没有细菌被摄入。然而，调理作用后，约7倍多的细菌位于细胞内，表明抗体促进吞噬作用。与此相反，细菌标记固有的GFP不能有效地吞噬，即使在调理抗体的存在下，表明FITC干扰细菌防御。由于FITC共价修饰蛋白质并可能影响其功能，我们测试了FITC对腺苷酸环化酶毒素活性（一种重要的细胞外毒力因子）的影响。FITC标记的细菌腺苷酸环化酶毒素活性比未标记的野生型细菌或GFP表达细菌低五倍，表明FITC损害腺苷酸环化酶毒素活性。这些数据表明，至少有一种细胞外毒力因子受到FITC标记的影响，GFP是一个更合适的标签为B。百日咳。
To explore the role of neutrophil phagocytosis in host defense againstBordetella pertussis, bacteria were labeled extrinsically with fluorescein isothiocyanate (FITC) or genetically with green fluorescent protein (GFP) and incubated with adherent human neutrophils in the presence or absence of heat-inactivated human immune serum. In the absence of antibodies, FITC-labeled bacteria were located primarily on the surface of the neutrophils with few bacteria ingested. However, after opsonization, about seven times more bacteria were located intracellularly, indicating that antibodies promoted phagocytosis. In contrast, bacteria labeled intrinsically with GFP were not efficiently phagocytosed even in the presence of opsonizing antibodies, suggesting that FITC interfered with a bacterial defense. Because FITC covalently modifies proteins and could affect their function, we tested the effect of FITC on adenylate cyclase toxin activity, an important extracellular virulence factor. FITC-labeled bacteria had fivefold-less adenylate cyclase toxin activity than did unlabeled wild-type bacteria or GFP-expressing bacteria, suggesting that FITC compromised adenylate cyclase toxin activity. These data demonstrated that at least one extracellular virulence factor was affected by FITC labeling and that GFP is a more appropriate label forB. pertussis.