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Mucus secretion studies using anti-gastric mucin monoclonal antibodies

使用抗胃粘蛋白单克隆抗体进行粘液分泌研究

基本信息

批准号：
07670612
负责人：
ISHIHARA Kazuhiko
金额：
$ 1.15万
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：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670612/
关键词：
anti-mucin antibody monoclonal antibody gastric mucin mucus secretion carbohydrate antigen 糖鐘抗原

项目摘要

Gastric mucin known as a major component of rat gastric mucus is synthesized in and secreted from several different kind of cells present at the mucosal epithelium. It is generally accepted that the regulation mechanism on the secretion and synthesis of mucin in a kind of cells present at the specific region and layr of gastric mucosa differ from the others. It is important to recognaize and estimate the specific mucin of gastric mucosa for understanding the role of mucus in the physiology and pathology of gastric mucosa. In this study we developed several monoclonal antibodies reacting specific mucin molecules and applied these antibodies to establish some methods to study the metabolism of gastric mucins.1) RGM11,21 and 24, are the monoclonal antibodies specifically react with the mucin present at the surface epithelial cells of rat gastric mucosa. It has been demonstrated that RGM11 and 21 recognize the specific oligosaccharide structure of mucin molecule. RGM24 reacts intact mucin molecules but the reactin disappears by the removal of saccharide residue from the mucin molecules.2) We developed 14 novel monoclonal antibodies (MAbs) using purified human colonic mucin as the immunogens (designated as HCM series MAbs). Among these MAbs, 8 MAbs are proved to recognize sialic acid-containing oligosaccaride structure of mucin molecule. In addition, some of these MAbs binding to sialic acid containing sugar chains are specifically react with the intestinal metaplastic mucosa in human stomach.3) We developed a sandwich ELISA mehtod using RGM11 and HIK1083 MAbs to recognize and quantify the surface mucous cell- and gland mucous cell-derived mucin, respectively, included in an extract of single mucosal specimen of rat stomach.

胃粘蛋白是大鼠胃粘液的主要成分，由胃粘膜上皮细胞合成和分泌。一般认为，胃粘膜特定区域和层内的一类细胞对粘蛋白分泌和合成的调节机制与其他细胞不同。胃粘膜特异性粘蛋白的定量分析和评价对于了解胃粘膜粘液在胃粘膜生理和病理中的作用具有重要意义。本研究开发了几种特异性的粘蛋白单克隆抗体，并应用这些抗体建立了一些研究胃粘蛋白代谢的方法。1）RGM11、21和24是与大鼠胃粘膜上皮细胞表面粘蛋白特异性反应的单克隆抗体。已经证明，RGM11和21识别粘蛋白分子的特异性寡糖结构。2）以纯化的人结肠粘蛋白为免疫原，制备了14株新的单克隆抗体（HCM系列单克隆抗体）。其中8株单克隆抗体可识别粘蛋白分子中含唾液酸的寡糖结构。另外，这些单克隆抗体中的一些与含唾液酸的糖链结合的单克隆抗体与人胃的肠化生粘膜特异性反应。3）我们分别用RGM11和HIK1083单克隆抗体建立了一种夹心ELISA方法来识别和定量大鼠胃粘膜单个标本提取物中的表面粘液细胞和腺粘液细胞来源的粘蛋白。