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）抗粘合蛋白单克隆抗体的发展和表征：我们将以下抗粘合蛋白单克隆抗体（MAB）集中在一起，并努力努力表征有关其表现体结构（a）指定为PGM34的MAB的信息，该MAB与酸性糖蛋白含有酸性糖含有硫酸糖的糖含量。 （b）一个称为RGM21的mAb，识别大鼠胃粘膜的表面粘液细胞衍生的粘蛋白与源自粘蛋白分子的相对较长的碳水化合物链反应。 c我们建立了29个mAb，针对源自人胃粘膜的粘蛋白。（2）在体外研究中估计幽门螺杆菌（HP）和人胃粘蛋白之间相互作用的估计：投资于HP对人类胃粘蛋白的粘附，我们开发了体内的实验性实验系统。 （a）可以通过表面等离子体共振分析的仪器观察到HP对纯化的人类胃粘蛋白的粘附。 （b）HP与纯化的ELISA纯化的人类胃粘蛋白反应。（3）通过使用抗粘膜蛋白MABS从实验性溃疡中愈合的大鼠胃粘膜的免疫组织化学观察结果：我们观察到使用抗Mucosa抗粘液蛋白的愈合阶段的粘蛋白种类的变化使用抗粘液蛋白疗法的变化。在胃粘膜的再生上皮中表达了特异性用RGM26和HCM31染色的粘蛋白。在急性胃粘膜损伤的愈合阶段（例如乙醇和阿司匹林诱导的溃疡）也获得了类似的观察结果。（4）将mABS应用于切除的人类消化道的病理观察：我们利用抗粘合蛋白mabs对人类gastric mucosae and cob癌的组织化学观察到抗粘液蛋白mab，并在抗胃肠炎的组织中进行了研究。事实证明，HGM75，HGM72，HCM14和HCM21对人类消化道的恶性变化的诊断和滞层有用。

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

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。

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。

Ichikawa T,Ishihara K,Hayashida H,Hiruma H,Saigenji K,Hotta K.: "Effect of ecabet sodium, a novel gastroprotective agent, on mucin metabolism in rat gastric mucosa."Digestive Diseases and Sciences. 45. 606-613 (2000)

Ichikawa T、Ishihara K、Hayashida H、Hiruma H、Saigenji K、Hotta K.：“新型胃保护剂依卡贝钠对大鼠胃粘膜粘蛋白代谢的影响。”消化疾病与科学。