Construction of human monoclonal antibodies for passive immuno therapy of oral diseases using Xenomouse

利用 Xenomouse 构建用于口腔疾病被动免疫治疗的人单克隆抗体

• 批准号: 13357017
• 负责人: ABIKO Yoshimitsu
• 金额: $ 27.71万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13357017/
• 关键词: Human monoclonal antibody; Xenomouse; Transchromo mouse; Passive immunization; Periodontal diseases; Porphyromonas gingivalis; Streptococcus sobrinus; Xeno mouse; transchromo-mouse; 赤血球凝集因子; 歯周病病原因子; ヘミン結合活性; ヒト型抗体; 不溶性グルカン; GTF; 口腔感染症; う蝕

The passive immunotherapy is a method for neutralizing the pathogenicity using antibody against each pathogen. Recently, it has been suggested that periodontal diseases are one of the consequential risk factors for several systemic diseases. As one of the medical treatments for the maintenance for human health, the development of passive immunotherapy for periodontal disease may be significant and necessary. Following research idea to construct the human monoclonal. antibodies (HmAbs) for the safety passive immunotherapy were carried out: Because the molecular information of the target antigens were important to use the HmAbs for passive immunotherapy, preliminary we attempted to clarify the molecule functional analysis of target antigens, such as hemagglutinin molecules and coaggregation factor of Porphyromonas gingivalis, and glucosyltransferases (GTFs) of Streptococcus sobrinus. Additionally, the large volume purification system by osmotic electrophoresis was also improved.The HmAbs … More for passive immunity examined 1) the human lymphocyte immortalization method using EB virus, 2) the method, using Xenomouse, 3) the method using Transchromomouse, and hmAbs were produced using these methods. In addition, recombinant ScFv using mouse monoclonal antibody producing hybridoma cell was improved into the large-scale production system, moreover, IgY antibody by the chicken immunity was also produced. In this study, the, practicable abilities of clones were screening using a Biacore system to detect the mAbs levels and avidity in cultured medium. The constructed HmAbs ability was determined the neutralization levels against the pathogenicity of antigens.Using Xenomouse technology, we succeeded to construct the HmAbs against the hemagglutinin of P. gingivalis, and GTFs of 」 sobrinus., Moreover, using Transchromomouse technology, HmAb against the coaggregation factor of P gingivalis was also constructed. Interestingly, the anti-coaggregation factor antibodies could increase the neutrophilic activity to phagocytize the P gingivlais cells. We also succeeded in the construction of IgY antibody against the hemagglutinin activity of P gingivalis. Less

被动免疫疗法是使用针对每种病原体的抗体来中和致病性的方法。最近，有人提出，牙周病是几种系统性疾病的后果的危险因素之一。作为维护人类健康的医学手段之一，牙周病被动免疫治疗的发展具有重要意义和必要性。按照研究思路构建人源单克隆抗体。由于靶抗原的分子信息对于HmAbs用于被动免疫治疗是重要的，因此我们初步尝试阐明靶抗原的分子功能分析，如牙龈卟啉单胞菌的血凝素分子和共聚集因子，以及远缘链球菌的葡萄糖基转移酶（GTF）。此外，还对渗透电泳大容量纯化系统进行了改进， ...更多信息 对于被动免疫，检查了1）使用EB病毒的人淋巴细胞永生化方法，2）使用Xenomouse的方法，3）使用Transchromomouse的方法，并使用这些方法产生hmAb。此外，利用鼠源性单克隆抗体杂交瘤细胞构建的重组ScFv已进入规模化生产体系，并通过鸡源性免疫制备了IgY抗体。本研究利用Biacore系统检测培养液中mAb的水平和亲和力，筛选克隆的实用能力。利用Xenomouse技术成功构建了抗牙龈卟啉单胞菌血凝素（HA）和牙龈卟啉单胞菌GTF（GTF）的HmAbs，此外，还利用转染色体小鼠技术，构建了抗牙龈卟啉单胞菌共聚集因子的HmAb。有趣的是，抗共聚集因子抗体可以增加嗜中性粒细胞吞噬牙龈卟啉单胞菌细胞的活性。成功地构建了抗牙龈卟啉单胞菌血凝素活性的IgY抗体。少

项目成果

M.Hayakawa: "Evaluation of the electroosmotic medium pump system for preparative disk gel electrophoresis"Anal.Biochem.. 288. 168-175 (2001)

M.Hayakawa：“用于制备盘凝胶电泳的电渗介质泵系统的评估”Anal.Biochem.. 288. 168-175 (2001)

Y.Otsuka: "Enhancement of plasminogen activator activity stimulated by LPS in gingival fibroblasts of individuals with down syndrome"J.Oral Sci.. 43. 207-212 (2001)

Y.Otsuka：“唐氏综合症个体牙龈成纤维细胞中 LPS 刺激的纤溶酶原激活剂活性的增强”J. Oral Sci.. 43. 207-212 (2001)

Y.Otsuka: "Enhancement of lipoplysaccharide-stimulated cyclooxygenase-2 mRNA expression and prostaglandin E2 production in gingival fibroblasts from individual with Down syndrome."Mecha.Age.Dev.. 123. 663-674 (2002)

Y.Otsuka：“唐氏综合症患者牙龈成纤维细胞中脂多糖刺激的环氧合酶 2 mRNA 表达和前列腺素 E2 生成的增强。”Mecha.Age.Dev.. 123. 663-674 (2002)

Y.Abiko: "Analysis of functional domains of Streptococcus sobrinus glucosyltransferase U."Int.J.Oral-Med.Sci.. 1. 152-156 (2003)

Y.Abiko：“远缘链球菌葡萄糖基转移酶 U 的功能域分析”Int.J.Oral-Med.Sci.. 1. 152-156 (2003)

M.Hayakawa, et al.: "Determination of short peptide in a Porphyromonas gingivalis protein antigen recognized by sera from periodontitis patients."Int.J.Oral-Med.Sci.. Vol.1,No.1. 79-81 (2002)

M.Hayakawa 等人：“牙周炎患者血清识别的牙龈卟啉单胞菌蛋白抗原中短肽的测定”。Int.J.Oral-Med.Sci. 第 1 卷，第 1 期。