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.
