Proteomic Profiling for Influenza Vaccination/Infection

描述(由申请人提供)：疫苗接种免疫反应的定量和定性分析是测试新疫苗以应对生物恐怖袭击的关键组成部分。此外，在发生新的/未知病原体的生物恐怖袭击时，及早发现和诊断感染源对于治疗和危机管理至关重要。同样，为了做好生物防御准备，还需要识别与病原体感染有关的新蛋白和宿主细胞对感染的反应。基于质谱学的蛋白质组学方法的最新进展可以应用于实现这些目标，这也是本提案的一个主要焦点。已经组建了一个强大的转化研究和临床团队，将新的蛋白质组和生物信息学工具与现有的免疫学分析相结合，既有流感疫苗平台，也有存档的受感染血清样本。我们的中心假设是，接种疫苗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源的反应是独特的，可以通过蛋白质组学来提取指纹。我们建议以C类生物恐怖病原体流感病毒为模型试剂，开发SELDI质谱仪蛋白质组图谱系统，用于监测疫苗反应和感染的早期检测/诊断。我们研究的最终目标是通过提高疫苗效力和早期诊断来减少自然和潜在的生物恐怖主义感染引起的流感的发病率和死亡率。将采取两种实验方法。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和接种后血清的蛋白质组图谱，以确定反映免疫反应的替代生物标志物。作为比较，类似的年轻人队列将接受鼻内活病毒FluMist疫苗。这些蛋白质谱的差异将与T细胞激活和对疫苗的抗体反应相关。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特征。在所有分析样本中发现的潜在生物标记蛋白将被分离出来，并通过质谱学进行测序。这些研究可能导致为检测、诊断和疫苗接种战略开发关键的新范例，这是加强我国针对病毒病原体的国家生物防御准备所必需的。

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.