Function of gastro-intestinal mucin-derived acidic oligosaccharaides recognized by anti-mucin monoclonal antibodies

抗粘蛋白单克隆抗体识别的胃肠粘蛋白衍生酸性寡糖的功能

• 批准号: 16590629
• 负责人: ISHIHARA Kazuhiko
• 金额: $ 2.11万
• 依托单位: Kitasato University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590629/
• 关键词: gastro-intestinal tract; mucin; mucosal damage; acidic oligosaccharide; 粘膜障害; モノクローナル抗体; 抗糖鎖抗体; スルホムチン; シアロムチン; 腸管寄生線虫

We developed a monoclonal antibody, designated PGM34, by immunizing a mouse with purified pig gastric mucin. PGM34 recognized the carbohydrate portion of the mucins. Oligosaccharide-alditols obtained from the pig gastric mucin were fractionated by successive gel-filtration, ion-exchange, and normal phase high-performance liquid chromatography, and tested for the reactivity with PGM34. Two purified oligosaccharide-alditols reacting with PGM34 were obtained and their structures were determined by nuclear magnetic resonance spectroscopy as Fucα1-2Galβ1-4GlcNAc(6SO_3H)β1-6(Fucα1-2Galβ1-3)GalNAc-ol and Fucα1-2-Galβ1-4GlcNAc (6SO_3H) β1-6 (Galβ1-3)Gal-NAc-ol. None of the defucosylated or desulfated form of these oligosaccharides reacted with PGM34. Thus, the epitope of PGM34 was determined as the Fucα1-2Galβ1-4GlcNAc (6SO_3H)β-sequence. Immunohistochemical observations of rat gastrointestinal tracts showed that PGM34 stained the surface mucous cells close to the generative cell zone in the g … More astric fundus and the goblet cells in the small intestine, but only slightly stained the antral mucous cells in the stomach. These data, taken together, showed that PGM34 is a very useful tool to reveal the role of mucins having characteristic sulfated oligosaccharides.To investigate the antineoplastic chemotherapy-induced small intestinal injury, 5-fluorouracil(5-FU) was orally administered to rats for 5 days. The 5-FU treatment caused a significant decrease in the number of mucus cells in the small intestine observed by an immunohistochemical method using PGM34. This reduction of mucus cells was significantly inhibited by the co-administration of lafutidine, a novel H2-blocker, but not by cimetidine, an H2-blocker of the first generation. The mucin content in the small intestine significantly decreased in the 5-FU treated group. This reduction was significantly recovered by the co-administration of lafutidine. These results support the efficacy of lafutidine toward 5-FU-induced mucositis in rat small intestine. Less

我们通过用纯化的猪胃粘蛋白免疫小鼠，开发了一种单克隆抗体，PGM34 识别从猪胃粘蛋白中获得的寡糖-糖醇，并通过连续的凝胶过滤、离子交换和分离。正相高效液相色谱法，并测试了两种纯化的寡糖-糖醇的反应性。获得与PGM34反应的产物，并通过核磁共振波谱确定其结构为Fucα1-2Galβ1-4GlcNAc(6SO_3H)β1-6(Fucα1-2Galβ1-3)GalNAc-ol和Fucα1-2-Galβ1-4GlcNAc(6SO_3H)β1- 6 (Galβ1-3)Gal-NAc-ol。这些寡糖的去岩藻糖基化或脱硫酸化形式与PGM34反应，因此，PGM34的表位被确定为Fucα1-2Galβ1-4GlcNAc(6SO_3H)β-序列。大鼠胃肠道的免疫组织化学观察表明，PGM34对靠近胃肠道的表面粘液细胞进行染色。胃底的生殖细胞区和这些数据综合起来表明，PGM34 是揭示具有特征性硫酸化寡糖的粘蛋白作用的非常有用的工具。诱导小肠损伤，给大鼠口服5-氟尿嘧啶（5-FU）5天，5-FU治疗引起粘液数量显着减少。使用 PGM34 的免疫组织化学方法观察到，粘液细胞的减少受到拉呋替丁（一种新型 H2 阻滞剂）的显着抑制，但西咪替丁（第一代粘蛋白阻滞剂）则没有显着抑制。在 5-FU 治疗组中，小肠中的含量显着降低，这种降低通过拉呋替丁的共同给药而显着恢复。拉呋替丁对 5-FU 诱导的大鼠小肠粘膜炎的影响较小。

项目成果

Effect of tea catechin administration on the change in gastro-intesttinal mucus in rats

茶儿茶素给药对大鼠胃肠粘液变化的影响

• 发表时间: 2005
• 期刊: WCOG Abstract, 13^<th> World Congress of Gastroenterology
• 作者: Ito Y;Ichikawa T;Ikezawa T;Saegusa Y;Tsubokawa D;Kuruhara M;Goso Y;Okayasu I;Ishihara K.
• 通讯作者: Ishihara K.

Characterization of oligosaccharide epitope recognized by anti-mucin monoclonal antibody, RGM21, using neoglycoconjugate technology

使用新糖缀合物技术表征抗粘蛋白单克隆抗体 RGM21 识别的寡糖表位

• 发表时间: 2005
• 期刊: Glycoconjugate Journal 22
• 作者: Goso Y;Endo M;Kuruhara M;Ishihara K.
• 通讯作者: Ishihara K.

Effects of a novel histamine H2-receptor antagonist, lafutidine, on the mucus barrier of human gastric mucosa

• DOI: 10.1111/j.1440-1746.2006.04721.x
• 发表时间: 2007-11-01
• 期刊: JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY
• 影响因子: 4.1
• 作者: Ichikawa, Takafumi;Ota, Hiroyoshi;Ishihara, Kazuhiko
• 通讯作者: Ishihara, Kazuhiko

Protective effects of the whisky congeners on ethanol-induced gastric mucosal damage

• DOI: 10.1111/j.1530-0277.2006.00319.x
• 发表时间: 2007-03-01
• 期刊: ALCOHOLISM-CLINICAL AND EXPERIMENTAL RESEARCH
• 影响因子: 3.2
• 作者: Goso, Yukinobu;Ueno, Masaki;Ishihara, Kazuhiko
• 通讯作者: Ishihara, Kazuhiko

Effect of Lafutidine, an H2-blocker, in the rat gastric mucosa regenetating from acetic acid-induced ulcer

H2阻滞剂拉呋替丁对醋酸溃疡大鼠胃粘膜再生的影响

• 发表时间: 2005
• 期刊: WCOG Abstract, 13^<th> World Congress of Gastroenterology
• 作者: Ikezawa T;Saegusa Y;Ichikawa T;Goso Y;Kuruhara M;Saigenji K;Okayasu I;Ishihara K.
• 通讯作者: Ishihara K.