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哺乳动物细胞上P菌毛特异性受体的密度与定位:荧光激活细胞分析

Density and localization of P-fimbriae-specific receptors on mammalian cells: fluorescence-activated cell analysis.

作者信息

Svenson S B, Källenius G

出版信息

Infection. 1983 Jan-Feb;11(1):6-12. doi: 10.1007/BF01651350.

Abstract

More than 95% of acute non-obstructive pyelonephritis cases in children are caused by P-fimbriated Escherichia coli. Uroepithelial cells from individuals prone to urinary tract infections bind such bacteria more avidly than do cells from healthy controls. A high density of receptors for P-fimbriae on cells of the urinary tract and kidney may thus be an important host factor in the acquisition of upper urinary tract infections. In this communication we are describing methods to determine the densities and localization of P-fimbriae receptors on mammalian cells. To determine the receptor density on cells in suspension, the cells were incubated with fluorescein-labelled P-fimbriated E. coli. They were then subjected to fluorescence-activated cell sorting (FACS) analysis which monitors both the size and the relative fluorescence of each cell. FACS analysis proved versatile for rapid and specific analyses of P-fimbriae receptor densities on large numbers of any chosen type of cell. The fluorescein-labelled P-fimbriated E. coli were also useful in localizing P-fimbriae receptors in tissues. These techniques are not limited to the assessment of P-fimbriae receptor densities and tissue localization but open up new aspects for studies of the specificities involved in the interactions between other microorganisms (e. g. parasites, bacteria and virus) and their host cells.

摘要

儿童急性非梗阻性肾盂肾炎病例中,超过95%由P菌毛化大肠杆菌引起。易患尿路感染的个体的尿路上皮细胞比健康对照者的细胞更易与这类细菌结合。因此,尿路和肾脏细胞上高密度的P菌毛受体可能是发生上尿路感染的一个重要宿主因素。在本通讯中,我们描述了测定哺乳动物细胞上P菌毛受体密度和定位的方法。为了测定悬浮细胞上的受体密度,将细胞与荧光素标记的P菌毛化大肠杆菌一起孵育。然后对它们进行荧光激活细胞分选(FACS)分析,该分析可监测每个细胞的大小和相对荧光。FACS分析被证明可用于快速、特异性地分析大量任何选定类型细胞上的P菌毛受体密度。荧光素标记的P菌毛化大肠杆菌在定位组织中的P菌毛受体方面也很有用。这些技术不仅限于评估P菌毛受体密度和组织定位,还为研究其他微生物(如寄生虫、细菌和病毒)与其宿主细胞之间相互作用的特异性开辟了新的领域。

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