Proteomic Profiling for Influenza Vaccination/Infection

描述（由申请人提供）：对疫苗接种的免疫反应进行定量和定性分析是为应对生物恐怖袭击而测试新疫苗的关键组成部分。此外，在发生新的/未知病原体的生物恐怖袭击时，早期发现和诊断感染因子对于治疗和危机管理至关重要。同样，鉴定与病原体感染和宿主细胞对感染的反应有关的新蛋白也需要进行生物防御准备。基于质谱的蛋白质组学方法的最新进展可以应用于实现这些目标，并且是本提案的主要焦点。一个强大的转化研究和临床团队已经组建，将新的蛋白质组学和生物信息学工具与现有的免疫学分析、流感疫苗平台和存档的感染血清样本结合起来。我们的中心假设是，对疫苗接种的免疫反应可以通过血清（或其他临床液体）的蛋白质组学分析来量化，并且宿主对不同感染因子的反应是独特的，可以通过蛋白质组学来“指纹化”。我们建议以C类生物恐怖主义病原体流感病毒为模型剂，建立SELDI质谱蛋白质组学分析系统，用于监测疫苗反应和早期发现/诊断感染。我们研究的最终目标是通过提高疫苗效力和早期诊断来降低自然和潜在的生物恐怖感染引起的流感发病率和死亡率。将采用两种实验方法。一种是使用从年轻和老年患者队列中获得的流感疫苗接种前后血清的蛋白质组学分析来确定反映免疫反应的替代生物标志物。为了进行比较，一个相似的年轻成人队列将接受活病毒鼻内流感疫苗。这些蛋白谱的差异将与T细胞活化和免疫应答相关。第二种方法将比较急性感染流感患者与对照组和rsv感染患者的血清和鼻拭子的蛋白质组学特征。所有分析样品中鉴定的潜在生物标记蛋白将通过质谱法分离和测序。这些研究可能导致关键的新范式的发展，以检测、诊断和疫苗接种战略，以增加我们国家对病毒性病原体的生物防御准备。

DESCRIPTION (provided by applicant): Quantitative and qualitative analysis of immune response to vaccination is a critical component for testing new vaccines in preparation for a bioterrorist attack. In addition, early detection and diagnosis of an infectious agent is crucial for treatment and crisis management in the event of a bioterrorist attack by a new/unknown pathogen. Similarly, identification of new proteins involved in pathogen infection and host-cell responses to infection are also needed for biodefense preparedness. Recent advances in mass spectrometry-based proteomic methods can be applied to achieve these goals, and is a major focus of this proposal. A strong translational research and clinical team has been assembled to combine new proteomic and bioinformatics tools with existing immunological assays, both influenza vaccine platforms, and archived infected serum samples. Our central hypothesis is that immune responses to vaccination can be quantified by proteomic profiling of serum (or other clinical fluids), and that the host response to different infectious agents are unique and can be 'fingerprinted' by proteomics. We propose to use influenza virus, a Category C bioterrorism pathogen, as a model agent to develop a SELDI mass spectrometry proteomic profiling system for monitoring vaccine response and early detection/diagnosis of infection. The ultimate goal of our study is to reduce the morbidity and mortality of influenza from natural and potential bioterrorism infections by improving vaccine efficacy and early diagnosis. Two experimental approaches will be taken. One is to use proteomic profiling of pre- and post influenza vaccination serum obtained from young and elderly patient cohorts to identify surrogate biomarkers reflective of the immune response. For comparison, a similar young adult cohort will receive the live-virus intranasal FluMist vaccine. These protein profile differences will be correlated with T cell activation and antibody responses to vaccination. The second approach will compare proteomic profiles from serum and nasal swabs of acutely infected influenza patients with control and RSV-infected patients. Potential biomarker proteins identified in all analyzed samples will be isolated and sequenced by mass spectrometry. These studies could lead to the development of crucial new paradigms for detection, diagnosis and vaccination strategies necessary to increase our national biodefense preparedness against viral pathogens.