Molecular Interactions in the Aging Immunological Synapse

衰老免疫突触中的分子相互作用

• 批准号: 7333193
• 负责人: NICHOLAS R GASCOIGNE
• 金额: $ 22.93万
• 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/7333193
• 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）：在衰老期间，淋巴细胞的发育和功能受到损害，从而导致年龄依赖性的感染剂易感性增加。 T细胞功能的下降是与衰老相关的免疫系统缺陷的最清晰和引人注目，从而导致免疫反应较低和较慢，这项研究将使用活细胞成像和Forster（或荧光）共振能量转移（FRET）显微镜来研究在粘液过程中触发体内的免疫学突触和分子相互作用的形成变化。在衰老期间，将研究TCR和共受体CD4或CD8之间相互作用的诱导。在老年或年轻TCR转基因动物的T细胞中，将研究免疫突触和TCR与共感染者之间的相互作用的形式。细胞将用基因转染以表达荧光嵌合蛋白，活细胞成像和FRET显微镜将用于分析TCR受体相互作用的诱导和动力学。可逆的蛋白质突出显示，具有光电可分离的荧光蛋白将用于测量细胞表面或突触中TCR或共肽的运动动力学。 T细胞需要识别自肽-MHC（PMHC）复合物，以通过限制抗原PMHC数量来帮助激活。将分析T细胞群体衰老期间此需求的变化。将测量T细胞对有限量抗原的敏感性的变化。用于脂质筏的荧光标记蛋白将用于比较年轻T细胞或老年T细胞中的筏动力学。 TCR和或共受体蛋白的运动将同时分析，以便筏动力学可以与突触的形成有关。可逆的蛋白质突出显示将用于可视化TCR或共感受器蛋白从木筏中和流出的运动。膜成分从抗原呈现细胞到T细胞的转移可能与年龄相关的变化（“ trogocytosis”）将被分析，以衡量T细胞膜性质的变化。免疫反应特别是T细胞随着年龄的增长而下降，使人更容易感染。该项目旨在确定在衰老过程中如何发生T细胞识别结构的变化，这对于了解如何预防与年龄相关的缺陷很重要。