利用流式细胞术和结合试验表征大肠杆菌热稳定肠毒素(STa)与其肠道推定受体在不同年龄组小鼠中的相互作用。

Laboratory animal science Pub Date : 1999-06-01
A M Al-Majali, J P Robinson, E K Asem, C Lamar, M J Freeman, A M Saeed
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引用次数: 0

摘要

背景与目的:产肠毒素大肠杆菌热稳定肠毒素(STa)是幼龄动物腹泻的主要原因。研究了STa特异性小鼠肠细胞受体的密度和亲和力的年龄依赖性变化。方法:选取2日龄、1、2周龄、2月龄4组瑞士韦氏小鼠(8 ~ 10只/组)。流式细胞术和放射性标记STa (125I-STa)检测被用作表征STa-肠细胞受体相互作用的可靠定量措施。结果和结论:STa与其推定受体的相互作用在2日龄小鼠肠细胞中较强。125i - sta受体相互作用的Scatchard分析表明,2日龄小鼠肠细胞中sta受体的数量(7.2 nmol/mg)高于老年小鼠(1周龄、2周龄和2月龄小鼠分别为0.30、0.36和0.40 nmol/mg)。此外,来自2日龄小鼠的受体对STa的亲和力(Kd = 75 nM)高于来自老年小鼠的受体(Kd分别为1周、2周和2月龄小鼠的125、1430和1111 nM)。肠细胞上STa受体的密度及其对STa的亲和力可能决定了STa的结合程度和分泌反应的严重程度,并可能解释了新生动物和人类婴儿对STa介导的腹泻的高易感性。
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Characterization of the interaction of Escherichia coli heat-stable enterotoxixn (STa) with its intestinal putative receptor in various age groups of mice, using flow cytometry and binding assays.

Background and purpose: Enterotoxigenic Escherichia coli heat-stable enterotoxin (STa) is a major cause of diarrhea in young animals. Age-dependent variation in the density and affinity of the mouse enterocyte receptors specific for STa was investigated.

Methods: Four age groups (2-day-, 1- and 2-week-, and 2-month-old) of Swiss Webster mice were studied (8 to 10 mice/group). Flow cytometry and radiolabeled STa (125I-STa) assays were used as reliable quantitative measures for characterization of STa-enterocyte receptor interaction.

Results and conclusions: Interaction of STa with its putative receptor was stronger for enterocytes of 2-day-old mice. Scatchard analysis of 125I-STa-receptor interaction suggested that STa-receptors exist at higher numbers on enterocytes from 2-day-old (7.2 nmol/mg) than older (0.30, 0.36, and 0.40 nmol/mg for 1-week-, 2-week-, and 2-month-old mice, respectively). Additionally, receptors from 2-day-old mice had greater affinity for STa (Kd = 75 nM) than did receptors from older mice (Kd = 125, 1,430, and 1,111 nM for 1-week-, 2-week-, and 2-month-old mice, respectively). Density of STa receptors on enterocytes and their affinity to STa may determine extent of binding and severity of the secretory response, and may explain the high susceptibility of newborn animals and human infants to STa-mediated diarrhea.

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