Fluorescent barcoding to unravel T cell niche development during vaccination.

荧光条形码可揭示疫苗接种过程中 T 细胞生态位的发育。

• 批准号: 9091409
• 负责人: Audrey Gerard
• 金额: $ 5.4万
• 依托单位: UNIVERSITY OF OXFORD
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9091409
• 关键词: Address; Affect; Affinity; Antigens; Behavior; CD8B1 gene; Cell Communication; Cells; Cellular Immunology; Characteristics; Clone Cells; Collection; Communicable Diseases; Coupled; Cues; Development; Disease; Environment; Equilibrium; Generations; Goals; Health; Image; Imagery; Imaging Techniques; Imaging technology; Immune; Immune response; Immune system; Immunity; Individual; Infection; Influenza; Investigation; Knowledge; Life; Longevity; Lymphoid; Maps; Memory; Methods; Molecular Immunology; Mutation; Organ; Output; Peptides; Phase; Physiological; Population; Process; Proteins; Resolution; Signal Transduction; Study Section; T cell differentiation; T cell regulation; T memory cell; T-Cell Activation; T-Lymphocyte; Techniques; Time; Vaccination; Virus; Virus Diseases; Work; base; career; cell behavior; cellular development; design; imaging modality; improved; in vivo; interest; long term memory; lymph nodes; novel; pathogen; response; single cell analysis; spatiotemporal; tool development; vaccine development

 DESCRIPTION (provided by applicant): Vaccination is one of the best examples of successful manipulation of the immune system. However, vaccination against pathogens like influenza that have a high mutation rate, display variable antigens, and require T cell immunity are still tenuous. The immune system is made up of a diverse collection of cells, each of which has distinct sets of triggers that elicit unique and overlapping responses. Effector, Memory and Tolerant states of CD8 T cells all emerge from a different set of multiple interactions regulated i space and time. Mapping those interactions during natural infection and vaccination and understanding which ones are regulating T cell fate are essential to understand how immune balance is maintained. We hypothesize that specific cell interactions regulate long-term protection and can be manipulated during vaccination. We and others have recently used live imaging to demonstrate that T cell priming takes place in the limited physical volume of the 'reactive' lymph node under highly dynamic conditions that would appear to permit considerable mixing of ongoing responses. A key finding is that there is a 'Critical Differentiation Period' tha coincides with individual activated T cell clones coming together forming a T cell niche environment. We propose to develop a new imaging technique based on fluorescent barcoding of individual T cell clones coupled with organ-clearing to increase our knowledge of the niche-dependent regulation of T cell fate. This method will be the first imaging technique to give access to single-cell behavior over time, as opposed to current methods characterizing the behavior of a population. It will rely on the expression of a unique set of fluorescent proteins in each cell within an immune population of interest. Using whole Lymph Nodes imaging coupled to fluorescent barcoding, we will specifically investigate how T cell niches are formed and for how long they persist. T cell niches are platforms where contact-dependent signals regulate CD8 T cell differentiation and so understanding interactions here will affect multiple disorders. This information will give novel information on CD8 T cell fate that will be critical to enhance vaccination against viruses.

 描述（由应用程序提供）：疫苗接种是成功操纵免疫系统的最佳例子之一。然而，针对具有高突变率，显示可变抗原的病原体的疫苗接种，并且需要T细胞免疫组织化合物仍然很脆弱。免疫系统由潜水细胞的集合组成，每个细胞都有不同的触发器集，这些触发因素引起了独特而重叠的响应。效应子，记忆和CD8 T细胞的耐受状态都来自不同的多个相互作用集的调节I空间和时间。在自然感染和疫苗期间映射这些相互作用，并了解哪些受调节的T细胞命运对于了解如何维持免疫平衡至关重要。我们假设特定的细胞相互作用调节了长期保护，并且可以在疫苗期间进行操纵。我们和其他人最近使用实时成像来证明T细胞启动发生在高度动态条件下的“反应性”淋巴结的有限物理体积中发生，这似乎可以考虑对正在进行的反应的考虑。一个关键的发现是，存在一个“关键的分化周期”，这与单个活化的T细胞克隆融合在一起形成T细胞生态位环境。我们建议开发一种基于单个T细胞克隆的荧光条形码和器官清除的新成像技术，以增加我们对T细胞命运的小裂依赖性调节的了解。与当前表征人口行为的当前方法相比，该方法将是第一种使单细胞行为访问单细胞行为的第一种成像技术。它将依赖于一组独特的荧光蛋白的表达 每个细胞在感兴趣的免疫人口中。使用与荧光条形码结合的全淋巴结成像，我们将专门研究如何形成T细胞壁ni，并持续多长时间。 T细胞壁ni是平台，在该平台中，接触依赖性信号调节CD8 T细胞分化，因此了解这里的相互作用将影响多种疾病。该信息将提供有关CD8 T细胞命运的新信息，这对于增强对病毒的疫苗接种至关重要。