Suppression of the Immune Response in Spaceflight and Aging

太空飞行和衰老过程中免疫反应的抑制

• 批准号: 8543621
• 负责人: MILLIE HUGHES-FULFORD
• 金额: $ 70.6万
• 依托单位: NORTHERN CALIFORNIA INSTITUTE/RES/EDU
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8543621
• 关键词: Affect; Aging; Altered Gravity; Astronauts; Bacterial Infections; Bioinformatics; CD4 Positive T Lymphocytes; Candidate Disease Gene; Clinical; Collection; Contracts; Data; Depressed mood; Disease; Double-Stranded RNA; Elderly; Environment; Enzyme-Linked Immunosorbent Assay; Epithelium; Fever Chills; Force of Gravity; Gel; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Goals; Head; Herpes zoster disease; Human; Humanities; Immune; Immune response; Immune system; Immunosuppression; In Vitro; Infection; Interleukin-2; International; Lymphocyte; Measures; MicroRNAs; Microarray Analysis; Microgravity; Mission; Modeling; Molecular; Ocular orbit; Older Population; Persons; Pharmacologic Substance; Phase; Production; Promoter Regions; Protein Biosynthesis; Proteins; Proteomics; Pseudomonas aeruginosa; Quantitative Reverse Transcriptase PCR; Quarantine; Regulation; Reporting; Research; Research Personnel; Signal Transduction Pathway; Simulate; Sodium Dodecyl Sulfate-PAGE; Space Flight; State of Zero Gravity; T cell response; T-Cell Activation; T-Cell Proliferation; T-Lymphocyte; Testing; Time; United States National Aeronautics and Space Administration; United States National Institutes of Health; Viral; Virus Diseases; Western Blotting; Work; age related; base; experience; immune function; in vivo; innovation; interest; novel; overexpression; pathogen; programs; research study; transcription factor

Project Summary In aging, immune systems falter and the elderly are more susceptible to infection. Age related changes include decreased T-cell activation/proliferation, decreased production of IL-2 and decreased expression of IL-2R¿. These are the same key immune changes that are seen in returning astronauts who have experienced altered immune function and increased vulnerability to infection during spaceflights. Loss of immune response in aging and spaceflight has been previously identified in T cells. We have previously identified microgravity induced changes in gene expression, promoter regions, transcription factors and signal transduction pathways in human CD4+ T-cells under normal and altered gravity conditions. Our preliminary data from International Space Station (ISS) described within this proposal show that in early T-cell activation at least one miRNA was upregulated during activation in normal gravity (g), while in microgravity (¿g), the induction of the miRNA was muted. Moreover, three protein targets of the miRNA had increased expression in the 1.g environment suggesting a micro RNA mechanism. The changes in immune function in the elderly occur over time and are hard to evaluate. However, the same type of changes in immune response occurs microgravity, both in astronauts and in human T-cells and is easy to measure The central hypothesis is that ¿g regulates MiRNA in human T-cells and that weightlessness is a novel model to study the immunosuppression seen in aging. These experiments will contribute to our fundamental understanding of the molecular mechanism of immunosuppression in spaceflight and in aging. The Specific Aims are the following: (1) Identify gene expression of miRNAs during T-cell activation (using arrays and qRTPCR) under normal gravity. (2) Identify the MiRNA(s) target genes using bioinformatics and verify the changes in expression of those targets messages (qRTPCR). We will test the ability of microgravity to induce/reduce expression of MiRNAs and their targets and verify these results using miRNA overexpression viral constructs or dsRNA. (3) Analyze the protein synthesis of the upregulated/downregulated target genes (SDS PAGE gels and Western Blot proteomics) that are affected by T-cell activation under normal gravity and microgravity conditions. (4) Compare the expression of key MiRNAs candidates and genes after activation in normal gravity and microgravity, vs. that of the lymphocytes from an older population.

项目摘要 随着年龄的增长，免疫系统会衰退，老年人更容易受到感染。年龄性变化 包括降低T细胞活化/增殖、降低IL-2产生和降低 IL-2 R的表达。这些是相同的关键免疫变化， 经历过免疫功能改变和感染脆弱性增加的宇航员 在太空飞行中。衰老和太空飞行中免疫反应的丧失此前已被证实 在T细胞中识别。我们以前已经确定了微重力引起的基因表达变化， 人CD 4 + T细胞的启动子区、转录因子和信号转导通路 在正常和变化的重力条件下。国际空间站的初步数据 (ISS)该提案中描述的研究表明，在早期T细胞活化中， 在正常重力（g）下激活期间上调，而在微重力（g）下， miRNA是沉默的。此外，miRNA的三个靶蛋白在1g/ml的小鼠中表达增加。 这表明了微RNA机制。老年人免疫功能的变化 随着时间的推移而发生，很难评估。然而，同样的免疫反应变化 发生微重力，无论是在宇航员和人类T细胞，是很容易测量的中央 一种假说认为，重力调节人类T细胞中的miRNA，而失重是一种新的模型， 研究衰老过程中的免疫抑制这些实验将有助于我们的基础 了解航天和衰老中免疫抑制的分子机制。 具体目的如下：（1）鉴定T细胞活化过程中miRNAs的基因表达 （使用阵列和qRTPCR）。(2)使用以下方法鉴定miRNA靶基因： 生物信息学和验证这些目标信息的表达变化（qRTPCR）。我们将测试 微重力诱导/减少MiRNA及其靶标表达的能力，并验证这些能力 结果使用miRNA过表达病毒构建体或dsRNA。(3)分析蛋白质合成 上调/下调的靶基因（SDS PAGE凝胶和蛋白质印迹蛋白质组学）， 在正常重力和微重力条件下受T细胞活化的影响。(4)比较 在正常重力和微重力下激活后关键的MiRNA候选物和基因的表达， vs.老年人的淋巴细胞。