LUMINEX TECHNOLOGY FOR THE QUANTIFICATION OF CYTOKINES IN NON-HUMAN PRIMATES

用于非人类灵长类动物细胞因子定量的 LUMINEX 技术

• 批准号: 8357650
• 负责人: Luis David Giavedoni
• 金额: $ 13.85万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357650
• 关键词: Aging; Animal Model; Animals; Antibodies; Callithrix; Cebidae; Cells; Cercopithecidae; Cercopithecus pygerythrus; Complex; Family suidae; Flow Cytometry; Funding; Genetic; Grant; Human; Immune system; Immunologics; Inflammation; Macaca; Macaca mulatta; Mediator of activation protein; Modeling; Monoclonal Antibodies; National Center for Research Resources; Obesity; Pan Genus; Papio; Pathogenesis; Physiological; Physiology; Play; Primates; Principal Investigator; Protocols documentation; Reagent; Recombinant Antibody; Recovery; Research; Research Infrastructure; Resources; Role; Scientist; Signal Transduction; Site; Source; System; Tail; Technology; Testing; Tube; United States National Institutes of Health; base; chemokine; cost; cytokine; improved; nonhuman primate; novel; pre-clinical research; vaccine development; web site

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Nonhuman primates (NHP) are the preferred animal models for pre-clinical research because they approximate humans in physiology and genetics more closely than any other animal. Critical advances in immunologic research have been made through the use of the NHP model, most notably in aids pathogenesis, treatment, and vaccine development. Cytokines and chemokines are soluble mediators of the immune system that play a crucial role in intercellular signaling, and in the recruitment of cells to inflammation sites. Identification of these molecules in NHP is crucial for the understanding of complex physiological and pathological mechanisms that occur in these species, and to demonstrate whether these mechanisms function similarly in humans. The luminex100 system is a bench-top flow cytometer that allows the user to quantify up to 100 molecules simultaneously in a single tube. Recently, we identified several antibodies specific for human cytokines that have the capacity to recognize homologous chemokines and cytokines of NHP origin. Currently, we have a panel of reagents that allow for the simultaneous identification of 23 cytokines and chemokines from chimpanzees and the old world monkeys rhesus macaques, baboons, cynomolgus macaques, pig-tailed macaques, and African green monkeys. in this r-24 application we propose to facilitate and expand the use of the Luminex technology in NHP by completing the following specific aims: 1) to improve the breadth and analytical power of our 23-plex NHP Luminex panel by increasing the number of detectable molecules in old world monkey species; 2) to improve the quality and depth of our 23-plex NHP Luminex panel by increasing the sensitivity of some suboptimal reagents currently used for quantification of cytokines from old world monkey species; and 3) to develop recombinant antibodies for the simultaneous quantification of cytokines in the marmoset, a new world monkey specie increasingly used in vaccine development, aging, and obesity research. We will perform extensive testing of available monoclonal antibodies against old world monkey cytokines, and state-of-the-art recombinant antibody technology for the creation of antibodies against marmoset cytokines. The information and protocols generated from these studies will be listed in the flow cytometry website of the SNPRC; novel antibodies will be distributed to scientists who justify their need on cost-recovery bases.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 非人类灵长类动物（NHP）是临床前研究的首选动物模型，因为它们比任何其他动物都更接近生理和遗传学的人类。 通过使用NHP模型，在AIDS发病机理，治疗和疫苗开发中，通过使用NHP模型来取得关键的进步。 细胞因子和趋化因子是免疫系统的可溶介质，在细胞间信号传导以及将细胞募集到炎症部位中起着至关重要的作用。 NHP中这些分子的鉴定对于理解这些物种中发生的复杂生理和病理机制至关重要，并证明了这些机制是否在人类中起作用。 Luminex100系统是一种台式流式细胞仪，允许用户在单个管中同时量化多达100个分子。 最近，我们确定了对人类细胞因子特异的几种抗体，它们具有识别NHP起源的同源趋化因子和细胞因子的能力。 目前，我们有一组试剂小组，可以同时鉴定来自黑猩猩和旧世界猴子鼠尾草猕猴的23种细胞因子和趋化因子，狒狒，cynomolgus猕猴，猪尾猕猴和非洲绿色猴子。 在此R-24应用中，我们建议通过完成以下特定目的来促进和扩展NHP中的Luminex技术的使用：1）通过增加旧世界猴子物种中可检测到的可检测到的分子的数量来提高我们23 plex NHP Luminex面板的宽度和分析能力； 2）通过提高目前用于定量旧世界猴子物种的细胞因子的某些次最佳试剂的敏感性来提高我们23- Plex NHP Luminex面板的质量和深度； 3）要开发重组抗体，以同时定量果酱中的细胞因子，这是一种新世界的猴子，越来越多地用于疫苗开发，衰老和肥胖研究。 我们将对针对旧世界猴子细胞因子的可用单克隆抗体以及最先进的重组抗体技术进行广泛的测试，以创建针对Marmoset细胞因子的抗体。 这些研究产生的信息和协议将在SNPRC的流式细胞仪网站上列出；新颖的抗体将分配给科学家，这些科学家证明其在成本恢复基础上的需求是合理的。