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

• 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
• 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

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.

项目总结 绒猴、松鼠猴和猫头鹰猴是被广泛研究的新大陆猴子(NWM) 广泛的研究领域，包括感染、视力、听力、社会行为、认知、 神经退行性疾病、生殖和基因操作。NWM的一个关键缺点 动物模型是目前可用的有限数量的免疫试剂，因为超过 3300万年的进化分化。研究工具的匮乏降低了翻译的价值 这些非人灵长类动物。这个R24应用程序的目标是通过生成 抗体和确认重要的NWM炎症和炎症生物标志物的免疫学分析 新陈代谢，目前缺乏试剂。炎症是衰老的常见过程， 肥胖，以及传染病、多发性硬化症和年龄相关的神经退行性变等病理疾病 紊乱，所有这些都是在这些物种中模拟的。将生产的NWM炎性生物标记物 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。会产生胃肠激素， 对代谢综合征研究的验证至关重要，包括胰岛素、脂联素和瘦素。为了 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，并建立了 生物技术公司，专业生产兽药和人类使用的试剂。我们会 在真核细胞中产生NWM蛋白，并用重组绒猴蛋白生产单克隆 小鼠的抗体(MMabs)(特异性目标1)。然后我们将使用绒猴重组蛋白来鉴定 这些单抗的表位特异性，筛选出还能识别松鼠猴和猫头猴的单抗 同源物，并确定最佳抗体对的技术，如ELISA，ELISPOT，Luminex和 细胞内细胞因子染色(ICS)分析(SA2)。最后，这些免疫学检测将通过 暴露于瞬时代谢和炎症的NWM血浆等分样本 挑战，以及来自体外刺激的NWM细胞(SA 3)。在这个项目结束时，研究人员在 人类疾病的不同NWM模型将可以使用研究工具来改进其有效性 临床前模型。将开发一个资源网站来传播试剂的可获得性，以跟踪 这些试剂在生物医学研究中的影响，以及分发方案和标准操作规程。这些方面的发展 试剂具有跨NIH的兴趣，将增加绒猴、松鼠猴和猫头鹰的翻译价值 猴子建立模型，并将扩展从这些动物获得的科学数据。