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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.

胃黏液蛋白是大鼠胃黏液的主要成分，是由存在于粘膜上皮的几种不同类型的细胞合成和分泌的。一般认为存在于胃粘膜特定区域和层的一类细胞对粘蛋白分泌和合成的调节机制不同于其他细胞。认识和估计胃粘膜特异性黏液蛋白，对于了解黏液在胃粘膜生理病理中的作用具有重要意义。本研究制备了几种与特异性粘蛋白分子反应的单克隆抗体，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是与存在于大鼠胃粘膜表面上皮细胞的粘蛋白特异性反应的单克隆抗体。研究表明，RGM11和rgm21能够识别粘蛋白分子的特异性低聚糖结构。RGM24反应完整的粘蛋白分子，但通过去除粘蛋白分子中的糖残基，反应蛋白消失。2)以纯化的人结肠粘蛋白为免疫原，制备了14种新型单克隆抗体（HCM系列单克隆抗体）。在这些单抗中，有8个单抗被证明能够识别黏蛋白分子中含唾液酸的低聚糖结构。此外，这些与含糖链唾液酸结合的单克隆抗体中，有一些与人胃肠化生黏膜发生特异性反应。3)采用RGM11和HIK1083单克隆抗体，建立了夹心ELISA方法，分别对大鼠胃粘膜标本提取液中的表面黏液细胞和腺体黏液细胞来源的黏液蛋白进行识别和定量。