Metabolic and Immune Responses to Flu Vaccine in Mitochondrial Disease Patients

描述(申请人提供)：在美国，每年有1000到4000名儿童出生时就患有线粒体疾病。由于症状可能出现在任何年龄，因此与接种疫苗的暂时联系也就不足为奇了。然而，对线粒体疾病患者的临床和疫苗免疫反应知之甚少。线粒体在T细胞激活中起重要作用；因此，我们建议对15名患有MELAS综合征的青少年和成人患者进行一项三价灭活流感疫苗(TIV)的试点研究，以描述引发的不良事件、详细的代谢反应和对TIV的免疫反应的深入评估。这些反应将与18-30岁的健康对照志愿者进行比较。所有受试者将在免疫前、免疫后6小时、5-7天和25-28天采集血液样本。在目标1中，将描述血液中线粒体功能的生物标记物，以确定TIV是否导致氧化应激。这将包括测定血浆中还原型谷胱甘肽(GSH)和氧化谷胱甘肽(GSSG)、有机酸、氨基酸、酰基肉碱和其他代谢物的GC和串联质谱学分析；FACS分析细胞内GSH、细胞内活性氧物种、细胞内活性氮物种和表面硫醇。在目标2中，将检查免疫反应，以评估特征是否随着线粒体功能障碍和年龄的变化而变化，哪些特征与抗体免疫反应强或差相关，以及抗体和T细胞反应是否受线粒体功能障碍的影响。这将通过使用Luminex系统分析血清中的42种细胞因子，使用Agilent 44K微阵列对RNA进行基因表达研究，通过细胞因子分析不同T细胞亚群中的淋巴细胞增殖，通过细胞因子测量流感特异性T细胞反应，以及使用磷酸盐分析多种细胞信号，来实现这一点。这项建议的独特之处在于，它首次研究了免疫对有氧化应激后代谢失代偿风险的患者代谢状态的影响，以及MELAS综合征是否影响T细胞和对TIV的抗体反应。对这些结果的分析将允许选择某些分析方法，这些方法可以在较年轻患者的后续研究中进行测试，因为较小的血容量限制了可以进行的分析次数。 公共卫生相关性：这项拟议的初步研究将提供重要的新信息，即与健康的年轻人对照组相比，患有MELAS综合征(一种线粒体疾病)的患者如何耐受流感疫苗免疫。对血液中生物标记物的专门研究将告诉我们，免疫是否会导致这些群体的氧化应激，如果是，这是否可能与不良事件有关。最后，对这些群体的免疫反应进行密集评估，将寻找预测流感疫苗保护性反应的生物标记物，以及可能受到这种线粒体紊乱影响的生物标记物。

DESCRIPTION (provided by applicant): Each year, 1000 to 4000 children in the U.S. are born with mitochondrial disease. Because symptoms may present at any age, it is not surprising that a temporal association with vaccination may occur. However, little is known about the clinical and immune response to vaccines in patients with mitochondrial disorders. Mitochondria play an important role in T cell activation; we therefore propose a pilot study of the administration of trivalent, inactivated influenza vaccine (TIV) to a group of 15 adolescent and adult patients with MELAS syndrome, a specific mitochondrial disorder, to describe solicited adverse events, detailed metabolic responses and an in-depth evaluation of immune responses to TIV. These responses will be compared with those of healthy control volunteers 18-30 yrs of age. Blood samples on all subjects will be taken prior to immunization, at 6h, 5-7 days and 25-28 days following immunization. In Aim 1, biomarkers of mitochondrial function in blood will be described to determine if TIV causes oxidative stress. This will include GC and tandem mass spectrometric assays for metabolomic profiling in plasma with measurement of reduced (GSH) and oxidized (GSSG) glutathione, organic acids, amino acids, acylcarnitines and other metabolites; FACS assays for intracellular GSH, intracellular reactive oxygen species, intracellular reactive nitrogen species and surface thiols. In Aim 2, immune response will be examined to assess if traits change with mitochondrial dysfunction and age, which traits correlate with a robust or poor antibody immune response and whether antibody and T cell responses are affected by mitochondrial dysfunction. This will be accomplished by analysis of 42 cytokines in serum using the Luminex system, white blood cell subsetting into 15 separate categories, gene expression studies performed on RNA using the Agilent 44K microarray, lymphocyte proliferation studies in the different T cell subsets with cytokine analysis of supernatants, influenza-specific T cell responses with measurement of cytokines, and multiplex cell signaling using phosphoflow assays. This proposal is unique in that it examines for the first time the effects of immunization on the metabolic status of patients who are at risk for metabolic decompensation following oxidative stress, and whether MELAS syndrome affects the T cell and antibody response to TIV. Analysis of these results will allow for selection of certain assays that can be tested in subsequent studies of younger patients where smaller blood volumes limit the number of assays that can be done. PUBLIC HEALTH RELEVANCE: This proposed pilot study will provide important new information about how patients with MELAS syndrome, a form of mitochondrial disease, tolerate immunization with flu vaccine in comparison with healthy young adult controls. Special studies of biomarkers in the blood will tell us if immunization causes oxidative stress in these groups and if so, whether this might be related to adverse events. Lastly, intense evaluation of immune responses in these groups will look for biomarkers that predict protective responses to influenza vaccine and that might be affected by this mitochondrial disorder.