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

被动免疫疗法是一种利用针对每个病原体的抗体中和致病性的方法。最近，有研究表明，牙周病是几种系统性疾病的危险因素之一。作为维护人类健康的医疗手段之一，牙周病被动免疫治疗的发展具有重要意义和必要性。遵循研究思路构建人的单克隆化。开展了用于安全被动免疫治疗的抗体(HmAbbs)：由于靶抗原的分子信息对于利用HmAbs进行被动免疫治疗非常重要，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，还对大容量渗透电泳法纯化系统进行了改进。HmAbs…被动免疫试验1)EB病毒人淋巴细胞永生化方法，2)异种小鼠永生化方法，3)转色小鼠方法，并用这些方法制备hmAb。此外，利用小鼠单抗生产杂交瘤细胞的重组ScFv被改进为规模化生产系统，此外，还利用鸡的免疫力生产了IgY抗体。本研究利用Biacore系统检测培养基单抗水平和亲和力，筛选出具有实用能力的克隆。利用异种鼠技术，成功地构建了抗牙龈假单胞菌血凝素的HmAbs和抗远缘青霉菌GTFs。此外，利用转色鼠技术，我们还成功地构建了抗牙龈假单胞菌共凝集因子的HmAb。有趣的是，抗共聚集因子抗体可以增加中性粒细胞吞噬牙龈假单胞菌细胞的活性。我们还成功地构建了抗牙龈假单胞菌血凝素活性的IgY抗体。较少

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