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

开发用于鉴定新世界猴生物标志物的免疫试剂

基本信息

批准号：
10592259
负责人：
Luis David Giavedoni
金额：
$ 49.77万
依托单位：
TRINITY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-15 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10592259
关键词：
Aging Aliquot Amino Acids Animal Model Animals Antibodies Aotus primate Area Behavior Binding Biological Biological Assay Biological Markers Biomedical Research Biotechnology Breeding C-reactive protein CXCL10 gene CXCR3 gene Callithrix Callithrix jacchus jacchus Cebidae Cells Cercopithecidae Chronic Disease Clinical Clinical Trials Cognition Communicable Diseases Data Development Enzyme-Linked Immunosorbent Assay Epitope Mapping Epitopes Eukaryotic Cell Exposure to Gastrointestinal Hormones Generations Genes Goals Granzyme Haplorhini Health Homologous Gene Hormones Human Immunologic Techniques Immunologics Immunology procedure Infection Inflammation Inflammatory Institution Insulin Interferon Type II Interleukin-10 Interleukin-4 Interleukin-6 Intervention Leptin Longevity Longitudinal Studies Macaca Macaca mulatta Malaria Membrane Glycoproteins Metabolic Metabolic hormone Metabolic syndrome Metabolism Modeling Monkeys Monoclonal Antibodies Multiple Sclerosis Mus National Institute of Allergy and Infectious Disease National Institute of Mental Health Neurodegenerative Disorders Neurology Neurosciences Night Monkey Obesity Outcome Pathology Plasma Pre-Clinical Model Primates Process Production Proteins Protocols documentation R24 Reagent Recombinant Proteins Recombinants Recommendation Reporting Reproduction Research Research Personnel Resources Rodent Saimiri Sampling Science Scientist Social Behavior South America South American Specificity Stains Techniques Technology Testing Transfection Transgenic Organisms United States United States National Academy of Sciences United States National Institutes of Health Validation Vision Western Blotting adiponectin age related neurodegeneration assay development biomarker identification cross reactivity cytokine enzyme linked immunospot assay genetic manipulation human disease human model improved interest murine monoclonal antibody nonhuman primate preclinical study tool validation studies web site

项目摘要

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.

项目摘要马尔莫果，松鼠猴和猫头鹰猴是新世界猴子（NWM）广泛研究各种各样的研究领域，包括感染，视觉，试听，社会行为，认知，神经退行性疾病，繁殖和遗传操作。 NWM的一个重要缺点动物模型是当前由于超过的免疫试剂数量有限 3300万年的进化差异。研究工具的这种稀缺性降低了这些非人类灵长类动物。该R24应用程序的目的是通过产生来解决此缺陷抗体和验证免疫学测定，鉴定出重要的NWM炎症和代谢，目前缺乏试剂。炎症是衰老涉及的常见过程，肥胖和病理，例如传染病，多发性硬化症和与年龄有关的神经退行性疾病疾病，所有这些都是在这些物种中建模的。生产的NWM炎症生物标志物包括C反应蛋白（CRP），白介素-4（IL-4），IL-6，IL-10，干扰素γ-诱导蛋白10 （CXCL10，IP-10），颗粒酶B和表面糖蛋白CD69。要产生的胃肠道激素，对于验证代谢综合征研究的至关重要，包括胰岛素，脂联素和瘦素。为了实现我们的目标，我们组建了一支来自学术和研究机构的科学家团队，生物技术公司专门生产兽医和人类用途的试剂。我们将在真核细胞中产生NWM蛋白质，并使用重组果胶蛋白产生单克隆小鼠中的抗体（MMAB）（特定目标1）。然后，我们将使用Marmoset重组蛋白来识别这些mAb的表位特异性，选择也识别松鼠猴和猫头鹰猴子的表位特异性同源物，并确定ELISA，ELISPOT，LUMINEX和细胞内细胞因子染色（ICS）测定（SA 2）。最后，这些免疫学测定将通过从暴露于瞬时代谢和炎症的NWM的血浆等分试样获得的样品挑战，从离体刺激NWM细胞（SA 3）。在该项目结束时，研究人员从事不同的NWM人类疾病模型将可以使用研究工具，以改善其验证临床前模型。将开发资源网站以传播试剂的可用性，以跟踪这些试剂在生物医学研究中的影响，并分发方案和SOP。这些的发展试剂具有trans-nih的兴趣，将增加马尔莫斯特，松鼠猴和猫头鹰的翻译价值猴子模块，并将扩大从这些动物获得的科学数据。