FLOW CYTOMETRIC ANALYSIS OF WITHIN-STRAIN VARIATION IN POLYSACCHARIDE EXPRESSION BY BACTEROIDES-FRAGILIS BY USE OF MURINE MONOCLONAL-ANTIBODIES
FLOW CYTOMETRIC ANALYSIS OF WITHIN-STRAIN VARIATION IN POLYSACCHARIDE EXPRESSION BY BACTEROIDES-FRAGILIS BY USE OF MURINE MONOCLONAL-ANTIBODIES
复制标题
DOI：
10.1099/00222615-35-4-229
复制
发表时间：
1991-10-01
影响因子：
3
通讯作者：
MCNEILL, TA
中科院分区：
文献类型：
--
作者：
LUTTON, DA;PATRICK, S;MCNEILL, TA
检测了4种不同单克隆抗体（MAbs）与脆弱拟杆菌（Bacteroides fragilis）NCTC 9343种群的反应性，所述脆弱拟杆菌NCTC 9343通过密度梯度离心富集为大胶囊、小胶囊和仅通过电子显微镜可见的电子致密层（EDL）。的单克隆抗体反应强烈，多糖存在于大胶囊和EDL丰富的人口，但不是在小胶囊丰富的人口。通过免疫印迹、免疫荧光和免疫电子显微镜以及流式细胞术确定标记模式。单克隆抗体标记的细胞膜相关的表位在大胶囊和EDL丰富的人口和细胞的材料在EDL人口。通过免疫印迹，在EDL群体中，但在大胶囊群体中，重复多糖亚基的梯状结构是明显的。通过流式细胞术测定每个群体内标记的细胞比例。通过流式细胞术证实另一种MAb与小囊群的反应性。表位表达的定性指标通过检查流式细胞术图谱获得。在结构不同的B之间和内部都观察到相同糖表位的差异表达。fragilis种群。这些单克隆抗体具有种特异性，并与最近的几种临床分离株发生交叉反应。这些多糖可能与B的毒力有关。脆弱的
The reactivity of four different monoclonal antibodies (MAbs) with populations of Bacteroides fragilis NCTC 9343, enriched by density gradient centrifugation for a large capsule, small capsule and electron-dense layer (EDL) only visible by electronmicroscopy, was examined. The MAbs reacted strongly with polysaccharides present in both the large capsule- and EDL-enriched populations but not in the small capsule-enriched populations. The pattern of labelling was determined by immunoblotting, immunofluorescence and immuno-electronmicroscopy, and flow cytometry. The MAbs labelled cell membrane-associated epitopes in the large capsule- and EDL-enriched populations and cell-free material in the EDL population. By immunoblotting, ladders of repeating polysaccharide subunits were evident in the EDL population but not in the large capsule population. The proportion of cells labelled within each population was determined by flow cytometry. The reactivity of another MAb with the small capsule population was confirmed by flow cytometry. A qualitative indication of epitope expression was obtained by examination of the flow cytometric profiles. Differential expression of the same saccharide epitope was observed both between and within structurally distinct B. fragilis populations. The MAbs were species-specific and cross-reacted with several recent clinical isolates. These polysaccharides may be relevant to the virulence of B. fragilis.