Development of Amygdala specific monoclonal Antibodies

描述（由申请人提供）：在本申请中，我们建议使用Vacold L.L.C公司建立的一种新型差异免疫方法“Differential Immunization for Antigen and Antibody Discovery”（DIAAD）技术，生成针对小鼠杏仁核特异性表位的大鼠单克隆抗体。DIAAD是由虚拟淋巴结（VLN）实现的，VLN是一种可植入的装置，作为一种强大的免疫佐剂，对微小剂量的抗原敏感。VLN在植入前装载的抗原剂量决定了免疫反应是否导致阳性免疫反应、能量或耐受。DIAAD技术对广泛的化学多样性抗原敏感，如脂类、碳水化合物、神经节苷类、蛋白质和翻译后蛋白修饰。因此，这项技术使我们能够耐受免疫系统对抗免疫显性和普遍表达的表位，并引导免疫系统从复杂的抗原混合物开始，转向免疫原性较低但组织特异性表达的抗原。

DESCRIPTION (provided by applicant): In this application we propose to use "Differential Immunization for Antigen and Antibody Discovery" (DIAAD) technology, a novel differential immunization method established by Vacold L.L.C., and generate rat monoclonal antibodies to epitopes specific to the Amygdala of mice. DIAAD is enabled by the Virtual Lymph Node (VLN), an implantable device used as a powerful immunological adjuvant with sensitivity to minute doses of antigen. The dose of antigen with which the VLN are loaded prior to implantation determines whether the immune reaction leads to a positive immune response, anergys or tolerance. The DIAAD technology is sensitive to antigens of broad chemical diversity, i.e., lipids, carbohydrate, gangliosides, proteins, and posttranslational protein modifications. Hence this technology allows us to tolerize the immune system against immuno-dominant and ubiquitously expressed epitopes and to direct the immune system towards less immunogenic but tissue specifically expressed antigens starting with complex mixtures of antigens. We chose the Amygdala as a target tissue since it is a central component of the structures, which consolidate and express emotions and has been implicated in the pathogenesis of affective mental disorders including depression. The possible importance of the Amygdala in the manifestation of affective mental disorders has been recognized for some time. Hence, there is a large scientific community both in the public and private sector, which devotes its resources to Amygdala research. One of the key objectives of the neuro-pharmaceutical industry is to develop novel therapeutics for the treatment of affective mental disorders. The specific aim of the proposed experiments is: to develop novel research tools for both applied and basic biomedical research in form of rat Monoclonal Antibodies (Moab's) against Amygdala specific epitopes of mice. The defined Milestone at the end of this phase 1 application is: to demonstrate the capacity to derive Amygdala specific MoAbs by differential tolerization /immunization using DIAAD and VLN technologies. These MoAbs should be usable for cytohistochemistry and/or western blot applications. We believe that these MoAbs will greatly accelerate research aimed to elucidate mechanisms of function of the Amygdala and will find widespread use in academic and industrial research. In a phase II extension of this application we wish to expand our portfolio of MoAbs reagents based on our results from phase I covering other important brain regions for human psychiatric diseases. In addition, we want to develop reagents to plastic changes of brain tissues in the context of inducible animal models with relevance to human psychiatric diseases.