Molecular Interactions in the Aging Immunological Synapse

衰老免疫突触中的分子相互作用

• 批准号: 7479289
• 负责人: NICHOLAS R GASCOIGNE
• 金额: $ 19.04万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7479289
• 关键词: Affect; Affinity; Age; Aging; Antibodies; Antigen-Presenting Cells; Antigens; Area; Binding; Biological Assay; CD4 Antigens; CD8B1 gene; Cell Aging; Cell membrane; Cell physiology; Cell surface; Cells; Ceramides; Chimeric Proteins; Chromosome Pairing; Data; Defect; Development; Employee Strikes; Fluorescence Recovery After Photobleaching; Fluorescence Resonance Energy Transfer; Genes; Immune response; Immune system; Individual; Infection; Infectious Agent; Kinetics; Left; Life; Ligands; Liquid substance; Lymphocyte; Measures; Membrane; Membrane Microdomains; Microscopy; Movement; Mus; Peptide/MHC Complex; Population; Predisposition; Property; Proteins; Staining method; Stains; Structure; Synapses; T-Lymphocyte; TCR Activation; Techniques; Time; Transgenic Animals; Transgenic Organisms; age related; aged; cell age; cellular imaging; immunological synapse; prevent; receptor; response; synaptogenesis

DESCRIPTION (provided by applicant): During aging, lymphocyte development and functions become compromised, resulting in age- dependent increases in susceptibility to infectious agents. The decline in T cell function is the most clear and striking of the immune system defects related to aging, resulting in lower and slower immune responses This study will use live cell imaging and Forster (or fluorescence) Resonance Energy Transfer (FRET) microscopy to investigate changes in formation of the immunological synapse and molecular interactions within the synapse during aging. The induction of the interaction between the TCR and co-receptor, CD4 or CD8, will be investigated during aging. The form of the immunological synapse and interactions between TCR and coreceptor within the synapse cells will be investigated in T cells from aged or young TCR transgenic animals. The cells will be transfected with genes to express fluorescent chimeric proteins and live cell imaging and FRET microscopy will be used to analyze the induction and kinetics of the TCR-coreceptor interaction. Reversible protein highlighting, with a photo-reactivatable fluorescent protein will be used to measure the movement kinetics of TCR or coreceptor on the cell surface or in the synapse. T cells require recognition of self-peptide-MHC (pMHC) complexes in order to aid in activation by limiting quantities of antigenic pMHC. Changes in this requirement during aging of the T cell population will be assayed. Changes in the sensitivity of the T cells to limited quantities of antigen will be measured. A fluorescent marker protein for lipid rafts will be used to compare raft dynamics in young or aged T cells. Movement of TCR and or co-receptor proteins will be analyzed at the same time, so that raft dynamics can be related to formation of the synapse. Reversible protein highlighting will be used to visualize movement of TCR or coreceptor proteins into and out of rafts. Possible age-related changes in the transfer of membrane components from the antigen presenting cell to the T cell ("trogocytosis") will be analyzed as a measure of changes in T cell membrane properties. The immune response particularly that of T cells, declines with age, leaving the individual more susceptible to infection. This project aims to determine how changes in the T cell's recognition structures occur during aging, which is important in understanding how age-related defects could be prevented.

描述（由申请人提供）：在衰老过程中，淋巴细胞的发育和功能受到损害，导致对传染性病原体的易感性随年龄增加。T细胞功能的下降是与衰老相关的免疫系统缺陷中最明显和最显著的，导致免疫反应降低和减慢。本研究将使用活细胞成像和福斯特（或荧光）共振能量转移（FRET）显微镜来研究衰老过程中免疫突触形成的变化和突触内的分子相互作用。TCR与辅助受体CD4或CD8之间相互作用的诱导将在衰老过程中进行研究。我们将在老龄和幼年TCR转基因动物的T细胞中研究免疫突触的形式以及突触细胞中TCR和辅受体之间的相互作用。细胞将被转染表达荧光嵌合蛋白的基因，活细胞成像和FRET显微镜将用于分析tcr -辅受体相互作用的诱导和动力学。可逆蛋白高亮，光活化荧光蛋白将用于测量TCR或辅助受体在细胞表面或突触内的运动动力学。T细胞需要自我肽- mhc （pMHC）复合物的识别，以便通过限制抗原pMHC的数量来帮助激活。在T细胞群的衰老过程中，这种需求的变化将被分析。将测量T细胞对有限数量抗原敏感性的变化。脂筏的荧光标记蛋白将用于比较年轻或老年T细胞的筏动力学。TCR和/或共受体蛋白的运动将同时被分析，因此筏动力学可以与突触的形成有关。可逆蛋白高光将用于可视化TCR或辅助受体蛋白进出筏的运动。细胞膜成分从抗原呈递细胞向T细胞转移过程中可能发生的与年龄相关的变化（“噬细胞症”）将作为T细胞膜特性变化的一种测量方法进行分析。免疫反应，尤其是T细胞的免疫反应，随着年龄的增长而下降，使个体更容易受到感染。该项目旨在确定T细胞识别结构的变化是如何在衰老过程中发生的，这对于理解如何预防与年龄相关的缺陷是很重要的。