Development of Immunological Reagents for the Identification of New World Monkey Biomarkers

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PROJECT SUMMARY Marmosets, squirrel monkeys, and owl monkeys are New World Monkeys (NWM) studied extensively in a broad array of research areas, including infection, vision, audition, social behavior, cognition, neurodegenerative diseases, reproduction, and genetic manipulation. One critical shortcoming of the NWM animal models is the limited number of immunological reagents that are currently available due to more than 33 million of years of evolutionary divergence. This scarcity of research tools reduces the translational value of these nonhuman primates. The goal of this R24 application is to remedy this deficiency by generating antibodies and validating immunological assays that identify important NWM biomarkers of inflammation and metabolism, for which there is currently lack of reagents. Inflammation is a common process involved in aging, obesity, and pathologies such as infectious diseases, multiple sclerosis, and age-related neurodegenerative disorders, all of which are being modeled in these species. The NWM inflammatory biomarkers to be produced include C-reactive protein (CRP), interleukin-4 (IL-4), IL-6, IL-10, interferon gamma-inducible protein 10 (CXCL10, IP-10), granzyme B, and the surface glycoprotein CD69. Gastrointestinal hormones to be produced, critical for validation of studies of metabolic syndrome, include insulin, adiponectin, and leptin. In order to accomplish our goal, we have assembled a team of scientists from academic and research institutions and a biotechnology company specialized in the production of reagents of veterinary and human use. We will produce NWM proteins in eukaryotic cells, and use the recombinant marmoset proteins to produce monoclonal antibodies (mMabs) in mice (Specific Aim 1). We will then use the marmoset recombinant proteins to identify epitope specificity of these mAbs, select the ones that also recognize squirrel monkey and owl monkey homologues, and identify optimal antibody pairs for techniques such as ELISA, ELISPOT, Luminex, and intracellular cytokine staining (ICS) assays (SA 2). Finally, these immunological assays will be validated with samples obtained from plasma aliquots from NWM exposed to transient metabolic and inflammatory challenges, and from ex vivo stimulated NWM cells (SA 3). At the end of this project, researchers working in different NWM models of human disease will have access to research tools for improved validation of their preclinical models. A resource website will be developed to disseminate the availability of reagents, to track the impact of these reagents in biomedical studies, and to distribute protocols and SOPs. Development of these reagents is of trans-NIH interest, will increase the translational value of marmoset, squirrel monkey and owl monkey modles, and will expand the scientific data obtained from these animals.