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.

项目概要 狨猴、松鼠猴和猫头鹰猴是新世界猴 (NWM)，在 广泛的研究领域，包括感染、视觉、听觉、社会行为、认知、 神经退行性疾病、生殖和基因操纵。 NWM 的一个关键缺点 动物模型是目前可用的免疫试剂数量有限的原因 3300万年的进化分歧。研究工具的缺乏降低了研究的转化价值 这些非人类灵长类动物。该 R24 应用程序的目标是通过生成来弥补这一缺陷 抗体和验证免疫学测定，识别重要的 NWM 炎症和生物标志物 代谢，目前缺乏试剂。炎症是衰老过程中的一个常见过程， 肥胖以及传染病、多发性硬化症和与年龄相关的神经退行性疾病等病症 所有这些疾病都在这些物种中进行建模。将产生的 NWM 炎症生物标志物 包括 C 反应蛋白 (CRP)、白细胞介素 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。产生胃肠激素， 对于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，我们聚集了一支来自学术和研究机构的科学家团队以及 专业生产兽用和人用试剂的生物技术公司。我们将 在真核细胞中产生 NWM 蛋白，并使用重组狨猴蛋白生产单克隆抗体 小鼠体内的抗体 (mMab)（具体目标 1）。然后我们将使用狨猴重组蛋白来鉴定 这些单克隆抗体的表位特异性，选择也识别松鼠猴和猫头鹰猴的单克隆抗体 同系物，并确定用于 ELISA、ELISPOT、Luminex 等技术的最佳抗体对， 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 从暴露于短暂代谢和炎症的 NWM 血浆等份中获得的样本 挑战，以及来自离体刺激的 NWM 细胞 (SA 3)。在该项目结束时，研究人员在 人类疾病的不同 NWM 模型将能够使用研究工具来改进其模型的验证 临床前模型。将开发一个资源网站来传播试剂的可用性，跟踪 这些试剂在生物医学研究中的影响，并分发方案和标准操作程序。开发这些 试剂具有跨 NIH 的兴趣，将增加狨猴、松鼠猴和猫头鹰的翻译价值 猴子模型，并将扩大从这些动物获得的科学数据。