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.