Miniaturized Automated Whole Blood Cellular Analysis System

小型化自动化全血细胞分析系统

• 批准号: 8935649
• 负责人: Angelle Desiree LaBeaud
• 金额: $ 39.83万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-26 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8935649
• 关键词: Affect; Antigens; Automation; Biological; Biological Assay; Blood; Blood Volume; Blood specimen; Cell physiology; Cells; Cellular Assay; Cellular Immunity; Child; Childhood; Clinic; Clinical; Clinical Research; Collection; Communicable Diseases; Competence; Cytometry; Data; Dengue Virus; Detection; Disease; Disease Progression; Elements; Equipment; Flow Cytometry; Freezing; Goals; Human; Immune; Immune response; Immunocompetence; Immunologic Monitoring; Individual; Infectious Agent; Kenya; Knowledge; Manuals; Measurement; Measures; Methods; Mitogens; Monitor; Pathogenesis; Peptides; Performance; Peripheral Blood Mononuclear Cell; Phenotype; Population; Preparation; Prevalence; Production; Proteomics; Protocols documentation; Qualifying; Reading; Reagent; Role; Running; Sampling; Serum; Shipping; Ships; Specificity; Specimen; Speed; System; Systems Analysis; T-Lymphocyte; Testing; Time; Tube; Universities; Vaccines; Viral; Virus; Whole Blood; Work; base; chikungunya; clinical research site; cohort; field study; immune function; innate immune function; instrument; miniaturize; pathogen; pediatric patients; point of care; prototype; public health relevance; response; single cell analysis; skills; vaccine development; vaccine trial

 DESCRIPTION (provided by applicant): Assays of cellular immunity are key to understanding the pathogenesis and mechanisms of control of viral and other infectious diseases. But such assays are difficult to perform as part of clinical studies because they are: º Labile: They must either be performed on fresh blood, or on PBMC that are cryopreserved within a relatively short time after blood collection. º Laborious: They require a lot of manual effort, as well as skills ad equipment not commonly found at clinical sites. º Sample-intensive: They tend to require large volumes of blood, particularly if performed on cryopreserved PBMC. These challenges have limited the implementation of cellular immune function assays, particularly in children (where blood draw volumes are most limited) and in remote settings. Yet, children in remote settings are often the most affected by important infectious diseases, and stand the most to gain from advances in vaccines and other methods of control. Therefore, we propose to develop a sample-sparing and fully automated system for stimulation and stabilization of whole blood for functional cellular assays. This system will be based on the concepts pioneered by Smart Tube, Inc. in their existing whole blood stimulation system, but will use 80% less blood, and provide full automation, so that all pipetting steps are eliminated. It will draw blood directly from a collection tube, using as little as 2 cc, distribute the blood into multiple incubation chambers where it will be stimulated with lyophilized, pre-configured reagents (antigens, mitogens, etc.) to assay cellular function, then stabilized with a proteomic stabilizer for later analysis. In Specifi Aim 1, we will develop this prototype system (through a subcontract with Smart Tube, Inc.) and test it on control samples in the Stanford Human Immune Monitoring Center. Two beta units will be built out for further testing. In Specific Aim 2, we will test these beta units at two clinical ites in Kenya through a subcontract with the Technical University of Mombasa. These clinical sites will collect blood samples from children infected with chikungunya and/or dengue virus. They will use the beta systems to stimulate and stabilize these blood samples, and ship them to Stanford for analysis by state-of-the art CyTOF mass cytometry. The results of this study will not only validate the blood collection system's performance, but will provide valuable biological data on the cellular response to these viruses in children. The blood collection system, in turn, will b useful for many different kinds of studies of cellular immunity, particularly in remote settings an situations where samples are limiting. The end result should be much more rapid advancement of our understanding of disease pathogenesis and of vaccine development for important infectious agents.

 描述（由适用提供）：细胞免疫学的测定是了解病毒和其他传染病控制的发病机理和机制的关键。但是，这样的评估很难作为临床研究的一部分进行，因为它们是：º阴性：它们必须在新鲜的血液上进行，或者在收集血液后相对较短的时间内进行冷冻保存的PBMC。 º费力：它们需要大量的手动努力以及在临床站点不常见的技能广告设备。 º样品密集型：它们倾向于需要大量的血液，尤其是在冷冻保存的PBMC上进行的。这些挑战限制了细胞免疫功能测定法的实施，尤其是在儿童（最有限的血量）和远程设置中。然而，偏远环境中的儿童通常受到重要感染疾病的影响最大，并且从疫苗和其他控制方法的进步中获得最大的收获。因此，我们建议开发一个样本的和完全自动化的系统，以刺激和稳定全血的功能性细胞测定。该系统将基于Smart Tube，Inc。在其现有的全血液刺激系统中开创的概念，但将使用80％的血液，并提供完整的自动化，以便消除所有移液步骤。它将直接从收集管中吸收血液，使用低至2 cc，将血液分配到多个孵化室中，在那里将用冻干的，预先配置的试剂（抗原，有差点等）刺激其刺激。 测定细胞功能，然后用蛋白质组学稳定剂稳定，以供以后分析。在Specifi AIM 1中，我们将开发此原型系统（通过Smart Tube，Inc。的分包合同），并在Stanford人类免疫监测中心的控制样品上对其进行测试。将建立两个Beta单元进行进一步测试。在Specifi AIM 2中，我们将通过与蒙巴萨技术大学的分包合同在肯尼亚的两个临床项目上测试这些β单元。这些临床部位将收集感染Chikungunya和/或登革热病毒的儿童的血液样本。他们将使用Beta系统刺激和稳定这些血液样本，并将其运送到斯坦福大学，通过最先进的质量细胞术进行分析。这项研究的结果不仅将验证血液收集系统的表现，而且还将提供有关儿童这些病毒的细胞反应的有价值的生物学数据。反过来，血液收集系统将对许多不同类型的细胞免疫学研究有用，尤其是在偏远环境中，样品限制了。最终结果应该更快地了解我们对疾病发病机理和重要感染剂疫苗开发的理解。