A systematic development of monoclonal antibodies against mucins of digestive tract and their application to pathophysiological studies

抗消化道粘蛋白单克隆抗体的系统开发及其在病理生理学研究中的应用

• 批准号: 12670514
• 负责人: ISHIHARA Kazuhiko
• 金额: $ 1.66万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670514/
• 关键词: mucin; monoclonal antibody; mucosal regeneration; experimental ulcer; stomach; immunohistochemistry; monoclonal antibody; eperimental ulcer

(1) Development and characterization of anti-mucin monoclonal antibodies: We focused the following anti-mucin monoclonal antibodies (mAbs) and made an effort to characterize the information about their epitome structure (a) An mAb designated as PGM34 reacted with acidic sugar moieties putatively bearing sulfated sugar residues. (b) An mAb named as RGM21 recognizing the surface mucous cell-derived mucin of rat gastric mucosa reacted with relatively long carbohydrate chain derived from the mucin molecules. c We established 29 mAbs against the mucin derived from human gastric mucosa.(2) Estimation of interaction between Helicobacter pylori (Hp) and human gastric mucin in in vitro study: To investgate the adhesion of Hp to human gastric mucin, we developed to in vitro experimental systems. (a) Adhesion of Hp to the purified human gastric mucin could be observed by an instrument of surface plasmon resonance analysis. (b) Hp reacted with the purified human gastric mucin pre-attached to the ELISA well.(3) Immunohistochernical observations of rat gastric mucosa healed from experimental ulcer by using anti-mucin mAbs: We observed the changes in the mucin species in the healing stage of rat gasric mucosa using anti-mucin mAbs developed in our laboratory. Mucins specifically stained with RGM26 and HCM31 was expressed in the regenerating epithelia of gastric mucosa. Similar observation was obtained in the healing stage of acute gastric mucosal injury such as ethanol- and aspirin-induced ulcer.(4) Application of the mAbs to the pathological observation of resected human digestive tract: We utilized anti-mucin mAbs to the histochemical observation of human gastric mucosae and colonic mucosae resected from the patient with digestive tract cancer, and several mAbs such as HGM75, HGM72, HCM14 and HCM21 proved to be useful to the diagnosis and clasification of the malignant changes in human digestive tract.

(1)抗粘蛋白单克隆抗体的开发和鉴定：我们重点研究了以下抗粘蛋白单克隆抗体（mAb），并努力表征了它们的epitome结构信息(a)一种被命名为PGM34的mAb与假定含有硫酸糖残基的酸性糖片段反应。(b)识别大鼠胃粘膜表面黏液细胞源性黏液蛋白的单克隆抗体RGM21与黏液蛋白分子衍生的较长碳水化合物链反应。c我们建立了29个针对人胃粘膜粘蛋白的单抗。(2)幽门螺杆菌（Helicobacter pylori, Hp）与人胃粘蛋白相互作用的体外研究：为了研究Hp与人胃粘蛋白的粘附性，我们开发了体外实验系统。(a)用表面等离子体共振分析仪可以观察到Hp与纯化的人胃粘蛋白的粘附。(b) Hp与预贴于ELISA的纯化人胃粘蛋白反应良好。(3)抗粘蛋白单抗对实验性溃疡愈合大鼠胃粘膜的免疫组化观察：利用本实验室研制的抗粘蛋白单抗，观察大鼠胃粘膜愈合阶段粘蛋白种类的变化。RGM26和HCM31特异性染色的粘蛋白在胃粘膜再生上皮中表达。急性胃粘膜损伤如乙醇性溃疡和阿司匹林性溃疡的愈合期也有类似的观察结果。(4)单克隆抗体在人消化道切除组织病理观察中的应用：我们利用抗粘蛋白单克隆抗体对消化道癌患者切除的人胃黏膜和结肠黏膜进行组织化学观察，HGM75、HGM72、HCM14、HCM21等单克隆抗体对人消化道恶性病变的诊断和分类有一定的帮助。

项目成果

Sawaguchi A, Ishihara K, kawano J, Oinuma T, Hotta K, Suganuma T.: "Fluid dynamics of the excretory flow of zymogenic and mucin contents in rat gastnc gland processed by high-pressure freezing/freeze substitution"The Journal of Histochemistry and Cytochem

Sawaguchi A、Ishihara K、kawano J、Oinuma T、Hotta K、Suganuma T.：“通过高压冷冻/冷冻替代处理的大鼠胃腺中酶原和粘蛋白内容物的排泄流的流体动力学”组织化学杂志和

Hayashida H, Ishihara K, Ichikawa T, Okayasu I, Kurihara M, Saigenji K, et al.: "Expression of a specific mucin type recognized by monoclonal antibodies in the rat gastric mucosa regenerating from acetic acid-induced ulcer"Scandinavian Journal of Gastroen

Hayashida H、Ishihara K、Ichikawa T、Okayasu I、Kurihara M、Saigenji K 等人：“乙酸诱发溃疡再生大鼠胃粘膜中单克隆抗体识别的特定粘蛋白类型的表达”斯堪的纳维亚胃病杂志

Hayashida H, Ishihara K, Ichikawa T, Okayasu I, Kurihara M, Saigeniji K, Hotta K: "Expression of a specific mucin type recognized by monoclonal antibodies in the rat gastric mucosa regenerating from acetic acid-induced ulcer"Scand J Grastroenterol. 36. 46

Hayashida H、Ishihara K、Ichikawa T、Okayasu I、Kurihara M、Saigeniji K、Hotta K：“从乙酸诱导的溃疡再生的大鼠胃粘膜中单克隆抗体识别的特定粘蛋白类型的表达”Scand J Grastroenterol。

Sawaguchi A, Ishihara K, Kawano J, Oinuma T, Hotta K, Suganuma T: "Fluid dynamics of the excretory flow of zymogenic and mucin contents in rat gastric gland processed by high pressure freezing/freeze substitutions"J Histchem Cytochem. 50. 223-234 (2002)

Sawaguchi A、Ishihara K、Kawano J、Oinuma T、Hotta K、Suganuma T：“通过高压冷冻/冷冻替代处理的大鼠胃腺中酶原和粘蛋白含量的排泄流的流体动力学”J Histchem Cytochem。

Obata F, Ichikawa T, Imai H, Nakagawa Y, Nakamura T, Ishihara K, et al.: "Selenium-Dependent Glutathione Peroxidase-GI Is Localized in Basal Granulated Cells of Rat Stomach"Biomedical Research. 22. 7-14 (2001)

Obata F、Ichikawa T、Imai H、Nakakawa Y、Nakamura T、Ishihara K 等人：“硒依赖性谷胱甘肽过氧化物酶-GI 位于大鼠胃的基底颗粒细胞中”生物医学研究。