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

被动免疫疗法是一种使用针对每种病原体的抗体中和致病性的方法。最近，有人提出牙周疾病是几种全身性疾病的结果风险因素之一。作为维持人类健康的医疗治疗方法之一，被动免疫疗法的牙周疾病可能是重要而必要的。遵循构建人类单克隆的研究思想。进行安全性被动免疫疗法的抗体（HMAB）：因为靶抗原的分子信息对于使用HMAB进行被动免疫疗法很重要，所以我们试图阐明靶标抗原的分子功能分析，例如脑血凝素分子和凝集素分子的孔酸盐剂孔隙蛋白酶，孔隙蛋白酶的孔隙蛋白酶，孔隙蛋白酶的孔隙蛋白酶孔的孔隙素化因素（gtfs）链球菌sobrinus。此外，还改善了通过渗透性电泳的大体积纯化系统。HMAB…更多用于使用EB病毒的被动免疫学检查1）使用Xenomouse，3）使用这些方法使用这些方法，使用Xenomouse，3）使用Xenomouse，3）使用这些方法，使用Xenomouse，3）使用这些方法，使用这些方法。此外，还将使用小鼠单克隆抗体产生杂交瘤细胞的重组SCFV改进到大规模生产系统中，此外，还产生了鸡肉免疫学的IGY抗体。在这项研究中，克隆的实际能力是使用Biacore系统筛选的，以检测培养基中的mAbs水平和亲和力。确定了针对抗原的致病性的神经化水平。使用异种构成技术，我们成功地构建了针对牙龈疟原虫的血凝素的HMAB，“ Sobrinus的GTF”和“ Sobrinus的GTF”。更换的，使用跨色素技术，Transchromome Technology，Hmab Ander the Gogegregation the Gogegregation with pogivalis of Gogivalis of Gingivalis of Gingivalis of Gingivalis of Gingivalis of Gogivalis。抗凝集因子抗体可以增加吞噬牙龈细胞的中性粒细胞活性

项目成果

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)

K.Teshirogi, et al.: "Production of monoclonal antibody inhibiting dipeptidylaminopeptidase IV activity of Porphyromonas gingivalis."Hybridoma Hybridomics. Vol.22,No.3. 147-151 (2003)

K.Teshirogi 等人：“抑制牙龈卟啉单胞菌二肽氨基肽酶 IV 活性的单克隆抗体的生产”。杂交​​瘤杂交组学。

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)

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。