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表征：我们重点关注以下抗粘蛋白单克隆抗体 (mAb)，并努力表征有关其表位结构的信息 (a) 命名为 PGM34 的 mAb 与酸性糖部分反应推测带有硫酸化糖残渣。 (b)名为RGM21的单克隆抗体识别大鼠胃粘膜表面粘液细胞衍生的粘蛋白，与粘蛋白分子衍生的相对较长的碳水化合物链发生反应。 c 我们建立了 29 种针对人胃粘膜来源的粘蛋白的单克隆抗体。(2) 在体外研究中评估幽门螺杆菌 (Hp) 与人胃粘蛋白之间的相互作用：为了研究 Hp 与人胃粘蛋白的粘附，我们开发了体外实验系统。 (a)可以通过表面等离子体共振分析仪器观察Hp与纯化的人胃粘蛋白的粘附。 (b) Hp 与预先附着在 ELISA 孔上的纯化人胃粘蛋白发生反应。(3) 使用抗粘蛋白单克隆抗体对实验性溃疡愈合的大鼠胃粘膜进行免疫组织化学观察：我们观察了愈合过程中粘蛋白种类的变化使用我们实验室开发的抗粘蛋白单克隆抗体对大鼠胃粘膜的阶段进行观察。用 RGM26 和 HCM31 特异性染色的粘蛋白在胃粘膜再生上皮中表达。在乙醇和阿司匹林引起的溃疡等急性胃粘膜损伤的愈合阶段也得到了类似的观察。 (4)单克隆抗体在切除的人体消化道病理观察中的应用：我们利用抗粘蛋白单克隆抗体进行组织化学观察从消化道癌患者身上切除的人胃粘膜和结肠粘膜，HGM75、HGM72、HCM14 和 HCM21 等几种单克隆抗体被证明有助于诊断人体消化道恶性变化的分类。

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。