Investigation of Neonatal Influenza Pathogenesis

新生儿流感发病机制的调查

• 批准号: 8850393
• 负责人: Alison J Carey
• 金额: $ 17.19万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8850393
• 关键词: Adult; Age; Alveolar Macrophages; Anti-Inflammatory Agents; Anti-inflammatory; Appointment; Biometry; CD8B1 gene; Cells; Cessation of life; Child; Clinical Services; Complex; Data; Death Rate; Disease; Dose; Elderly; Environment; Epithelial Cells; Exhibits; Face; Funding; Gene Activation; Gene Expression Profile; Goals; Green Fluorescent Proteins; Health; Hospitalization; Host Defense; Human; Immune; Immune response; Immune system; Immunity; Immunology; Infant; Infection; Inflammation Mediators; Inflammatory Response; Influenza; Intranasal Administration; Investigation; K-Series Research Career Programs; Knowledge; Laboratories; Lactobacillus; Lactobacillus casei rhamnosus; Lead; Left; Life; Light; Literature; Lower respiratory tract structure; Lung; Medicine; Mentors; Microbiology; Modeling; Mononuclear; Morbidity - disease rate; Mus; Neonatal; Neonatal Mortality; Outcome; Pathogenesis; Patients; Pediatrics; Phagocytes; Play; Population; Positioning Attribute; Predisposition; Premature Infant; Probiotics; Process; Property; Public Health; Pulmonary alveolar structure; RNA; Recombinants; Research; Research Personnel; Research Project Grants; Research Proposals; Resources; Respiratory Tract Infections; Role; Safety; Schools; Severities; Solutions; Sorting - Cell Movement; T cell response; T-Lymphocyte; Therapeutic; Therapeutic Intervention; Time; Training; Translational Research; United States National Institutes of Health; Universities; Vaccines; Viral; Viral Load result; Viral Pathogenesis; Virus; Virus Diseases; Work; World Health Organization; age group; base; career; clinically relevant; college; cytokine; driving force; fighting; fluorescence activated cell sorter device; in vivo; influenza virus vaccine; influenzavirus; meetings; microbiome; mortality; mouse model; neonate; next generation sequencing; novel; novel therapeutics; prevent; professor; pup; respiratory; respiratory virus; response; skills; success; therapeutic target; transcriptome sequencing

DESCRIPTION (provided by applicant): Dr. Carey is a Neonatologist and has a dual appointment as an Assistant Professor of Pediatrics and Microbiology/Immunology at Drexel University College of Medicine. Her current investigative efforts are directed towards elucidating the pathogenic mechanisms underlying morbidity and mortality of respiratory viral infections in the preterm infant. Neonates face one of the highest rates of mortality and morbidity from infection. According to the World Health Organization, 3.7 million neonates younger than 28 days of age died in 2010, and 37% of these deaths were due to infectious causes1. Specifically, influenza infection leads to many hospitalizations and deaths each year, mainly in children and the elderly. There are little data on why neonates exhibit increased mortality to influenza virus, although we know they do not have the mature, full immune repertoire that adults have at their disposal. In order to investigate this increased mortality, we have established a clinically-relevant model of neonatal influenza virus infection in 3-day old neonatal mice. We hope to elucidate mechanisms of increased pathogenesis in order to develop therapeutic interventions for an extremely vulnerable, but understudied, population. In our preliminary work, we determined that when 3-day old mice are infected with influenza, there is a high rate of death, approaching 80% by 8 days post infection. The amount of virus in the pup's lungs is higher immediately after infection as compared to adults. This early loss of viral control cannot be attributed to viral- specific T-cell immunity, as it is prior to the period when adults or neonates mount an immunodominant viral- specific CD8+ T-cell response in the lungs. This suggests that the lung innate host response in neonates is qualitatively different from that of adults. By using our novel neonatal mouse model, we are proposing studies to determine the critical differences in how the neonatal mice respond to influenza virus infection. As we investigate the situation further, we propose two research goals: 1) Determine differences in gene activation in infected neonatal mouse lung cells versus adult mouse lung cells. By using a novel recombinant Influenza virus that expresses Green Fluorescent Protein (PR8-GFP) and transcriptome analysis by Next Generation Sequencing of RNA (RNAseq), we will examine whether neonatal lung epithelial cells and alveolar macrophages respond differently to infection with influenza virus than adult cells. 2) Determine whether providing a probiotic will help protect neonatal mice from influenza infection. Based on its effects on adult respiratory infection, we hypothesize that microbiome colonization with probiotics like Lactobacillus can alter immunity to respiratory viral infection and may serve as a potential therapy to help either prevent or diminish the pathogenesis of respiratory viral infections in infants. However, up to this point, there have not been investigations into the use of intranasal probiotics for respiratory infections in infants. Attaining more knowledge of neonatal immune responses is essential if we are to develop effective vaccines or therapeutics against influenza virus or other respiratory viruses. This knowledge may allow us to develop novel therapeutic strategies against viral respiratory infections that reduce morbidity and mortality in neonates. This research proposal represents a critical step in attaining the candidate's long-term goal: to transition to an independent research career focused on mechanistic studies of respiratory viral pathogenesis in the preterm infant. In addition to the proposed research, Dr. Carey has formulated a multi- pronged training plan, including: didactic courses, scientific meetings, clinical service, and a superb mentoring team. This plan will build her skills in biostatistics and immunology, hone her critical review of the scientific literature, develop her ability to present her work in both the domestic and internationl arena, and acquire the ability to manage a laboratory. Continued access to the abundant resources and rich intellectual environment across departments and schools at Drexel University leave this candidate well-positioned to achieve this goal. This Career Development Award will serve as a driving force in Dr. Carey's success, providing the protected time and additional training required for this candidate to become an independent investigator.

描述（由申请人提供）：凯里博士是新生儿学家，并在德雷克塞尔大学医学院的儿科和微生物学/免疫学助理教授任命双重任命。她目前的调查工作旨在阐明早产儿中呼吸道病毒感染的发病率和死亡率的致病机制。新生儿面临着感染的死亡率和发病率最高的人之一。根据世界卫生组织的数据，2010年28天以下的370万新生儿死亡，其中37％的死亡是由于传染性原因1。具体而言，流感感染每年都会导致许多住院和死亡，主要是儿童和老年人。关于为什么新生儿对流感病毒的死亡率增加的数据很少，尽管我们知道它们没有成年人可以使用的成熟，完全免疫曲目。为了调查这种增加的死亡率，我们已经建立了3天老新生小鼠中新生儿流感病毒感染的临床相关模型。我们希望阐明发病机理增加的机制，以开发出极为脆弱但经过研究的人群的治疗干预措施。 在我们的初步工作中，我们确定3天老鼠感染了流感时，死亡率很高，感染后8天接近80％。与成年人相比，感染后幼犬肺中的病毒量更高。病毒控制的早期丧失不能归因于病毒特异性T细胞免疫，因为它是在成年人或新生儿之前 在肺中安装免疫主导的病毒 - 特异性CD8+ T细胞反应。这表明新生儿的肺与生俱来的宿主反应在质上不同于成年人。通过使用我们新的新生小鼠模型，我们提出研究以确定新生小鼠对流感病毒感染的反应方式的关键差异。随着我们进一步研究情况，我们提出了两个研究目标：1）确定受感染的新生小鼠肺细胞与成年小鼠肺细胞的基因激活差异。 通过使用一种新型的重组流感病毒，该病毒通过RNA的下一代测序（RNASEQ）表达绿色荧光蛋白（PR8-GFP）和转录组分析，我们将检查新生儿肺上皮细胞和肺泡巨噬细胞对对流感病毒细胞的感染是否不同。 2）确定提供益生菌是否有助于保护新生小鼠 来自流感感染。根据其对成人呼吸道感染的影响，我们假设用益生菌等微生物组定植可以改变对呼吸道病毒感染的免疫力，并可能是帮助预防或减少婴儿呼吸道病毒感染的发病机理的潜在疗法。但是，到目前为止，还没有研究使用鼻内益生菌用于婴儿的呼吸道感染。 如果我们要针对流感病毒或其他呼吸道病毒开发有效的疫苗或治疗剂，那么了解新生儿免疫反应的更多知识至关重要。这些知识可能使我们能够针对病毒呼吸道感染制定新的治疗策略，以降低新生儿的发病率和死亡率。这项研究建议是实现候选人的长期目标的关键步骤：过渡到独立研究 职业专注于早产婴儿呼吸道病毒发病机理的机理研究。除了拟议的研究外，凯里博士还制定了多项培训计划，包括：教学课程，科学会议，临床服务和一支精湛的指导团队。该计划将在生物统计学和免疫学方面提高她的技能，磨练她对科学文献的批判性审查，增强她在国内和国际领域介绍自己的作品的能力，并获得管理实验室的能力。在德雷克塞尔大学（Drexel University）的部门和学校之间继续获得丰富的资源和丰富的知识环境，使这位候选人置于实现这一目标。该职业发展奖将成为凯里博士成功的推动力，为该候选人成为独立调查员所需的受保护时间和额外的培训。