Binding of Norovirus Virus-Like Particles (VLPs) to Human Intestinal Caco-2 Cells and the Suppressive Effect of Pasteurized Bovine Colostrum on This VLP Binding
Binding of Norovirus Virus-Like Particles (VLPs) to Human Intestinal Caco-2 Cells and the Suppressive Effect of Pasteurized Bovine Colostrum on This VLP Binding
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DOI：
10.1271/bbb.90729
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发表时间：
2010-03-01
影响因子：
1.6
通讯作者：
Matsuda, Tsukasa
中科院分区：
文献类型：
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作者：
Murakami, Kosuke;Suzuki, Sayaka;Matsuda, Tsukasa
诺如病毒(NOV)不能在细胞培养中生长，是胃肠炎的主要感染源。建立了一种利用杆状病毒系统中产生的病毒样颗粒(VLP)来评价NOV与肠细胞结合的体外检测系统。在MS分析证实纯度后，将VLP与人肠道Caco-2细胞孵育。共聚焦激光显微镜仅在一定数量的Caco-2细胞上清楚地检测到NOV VLP，并通过细胞裂解产物的免疫印迹进行半定量。免疫印迹法检测巴氏杀菌牛初乳对VLP与Caco-2细胞结合的抑制作用。初乳以剂量依赖的方式减少VLP结合，12.5mg初乳蛋白抑制约50%。此外，初乳中含有与VLP反应的抗体，这表明牛初乳中的交叉反应抗体阻止了人NOV与肠道细胞的结合。
Noroviruses (NoVs), which cannot be grown in cell culture, are a major infectious agent of gastroenteritis. An in vitro assay system was established for the evaluation of NoV binding to enterocytes using virus-like particles (VLPs) produced in a baculovirus system expressing a NoV VP1 capsid protein. After confirmation of the purity by MS analysis, VLPs were incubated with human intestinal Caco-2 cells. NoV VLPs were detected clearly by confocal laser microscopy only on a certain population of Caco-2 cells, and were semi-quantified by immunoblotting of cell lysates. Then the suppressive effect of pasteurized bovine colostrum was analyzed on the VLP binding to Caco-2 cells by immunoblotting. The colostrum reduced VLP binding in a dose-dependent manner, at about 50% suppression with 12.5 mu g of the colostral proteins. Furthermore, the colostrum contained IgG antibodies reacting to VLPs, suggesting that cross-reactive antibodies in the bovine colostrums block human NoV binding to intestinal cells.