Mucosal and Systemic Immune Responses to Influenza Virus

对流感病毒的粘膜和全身免疫反应

• 批准号: 9188802
• 负责人: Harry Bernard Greenberg
• 金额: $ 42.79万
• 依托单位: PALO ALTO VETERANS INSTIT FOR RESEARCH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9188802
• 关键词: Acute; Address; Adult; Age-Years; Antibodies; Antibody Formation; Antibody Response; Attenuated; Avian Influenza; B-Lymphocytes; Blood; Blood Cells; Blood specimen; Cells; Child; Clinical; Clinical Research; Collaborations; Communities; Data; Development; Epithelial; Epithelial Cells; Extramural Activities; Flow Cytometry; Flu virus; Gene Expression Profile; Genetic Transcription; Genome; Goals; Healthcare Systems; Human; Humoral Immunities; Immune; Immune response; Immune system; Immunity; Immunization; Immunologic Markers; Individual; Infection; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H5N1 Subtype; Influenza A Virus, H7N9 Subtype; Influenza A virus; Influenza vaccination; Innate Immune Response; Innate Immune System; Knowledge; Leukocytes; Lymphocyte; Measurement; Measures; Microfluidics; Mucosal Immune Responses; Mucous Membrane; Nasal Epithelium; National Institute of Allergy and Infectious Disease; Natural Immunity; Nose; Pathogenicity; Patients; Peripheral; Plasmablast; Population; Quantitative Reverse Transcriptase PCR; Research; Research Personnel; Route; Sampling; Serological; Serum; Site; Structure of mucous membrane of nose; Study Subject; Swab; System; Technology; Testing; Time; United States National Institutes of Health; Upper respiratory tract; Vaccines; Viral; Virulence; Virus; Virus Diseases; Virus Replication; adaptive immune response; adaptive immunity; cell type; fighting; flu; healthy volunteer; immunogenicity; improved; influenza virus vaccine; influenzavirus; live attenuated influenza vaccine; novel; pandemic disease; pandemic influenza; predictive marker; protective efficacy; public health relevance; response; response biomarker; seasonal influenza

DESCRIPTION (provided by applicant): The 2009 influenza (flu) pandemic caused by the A(H1N1)pdm09 virus and the sporadic human cases of highly pathogenic avian flu A virus infection emphasize the need for improved flu vaccines for both seasonal and potential pandemic flu strains. One approach to flu vaccination entails the nasal administration of live attenuated flu viruses (LAIV), which mimics, in part, the natural route and site of viral infectionin the human nasal epithelial cells (HNEC) of the upper respiratory tract. However, LAIV fail to induce a robust systemic immune response compared to natural wild-type (wt) flu infections, suggesting important differences in the host innate and adaptive immune responses to attenuated and wt viruses. Flu virus replication in HNEC results in the induction of host immune responses within both epithelial and leukocyte cell subsets of the nasal mucosa. On the other hand, flu viruses encode multiple strategies to regulate and suppress the initial host innate response in both a strain- and cell type-specific manner, which is an important determinant of flu pathogenicity. We hypothesize that measurement of (a) early host transcriptional responses to LAIV vs. wt strains in HNEC as well as in mucosal and peripheral lymphocyte populations and (b) peripheral plasmablast B cell and serum and nasal antibody responses from the same individuals will identify specific early immune markers predictive of viral replication, immunogenicity, and ultimately protection. We propose to take advantage of an ongoing flu A(H1N1)pdm09 challenge study at the NIH Clinical Center to test this hypothesis. We will collect two additional nasal swabs from study subjects and sort nasal epithelial and lymphocyte cell populations by flow cytometry for subsequent high-throughput microfluidic qRT-PCR analysis of mucosal immune responses and local viral replication. Blood and nasal wash samples are also being collected from the same individuals to measure peripheral transcriptional responses and systemic B cell and antibody responses. The specific aims are to: 1) Characterize early host immune transcriptional responses in bulk unsorted nasal mucosal cells and sorted HNEC and nasal lymphocyte populations and in peripheral blood cells, to identify common and unique transcriptional signatures induced by wt virus and compare them to the host responses to LAIV immunization, which is being studied with similar approaches in other ongoing studies at Stanford; and 2) Characterize peripheral plasmablast B cell and serum and mucosal antibody responses to wt flu infection at a quantitative level in these same individuals and compare these to other ongoing studies of LAIV at Stanford, to identify local and/or systemic early transcriptional innate immunity response markers that correlate with subsequent plasmablast and other adaptive local and systemic humoral immunity indicators. These studies are expected to generate new knowledge on the underlying mechanism for protective immunity against flu viruses, which will guide the development of improved seasonal and pandemic flu vaccines.

描述（由申请人提供）：2009 年由 A(H1N1)pdm09 病毒引起的流感大流行和高致病性禽流感 A 病毒感染的散发人类病例强调需要针对季节性和潜在大流行性流感改进流感疫苗菌株。流感疫苗接种的一种方法是鼻腔注射减毒流感病毒（LAIV），它部分模仿了上呼吸道人类鼻上皮细胞（HNEC）病毒感染的自然途径和部位。然而，与自然野生型（wt）流感感染相比，LAIV 未能诱导强大的全身免疫反应，这表明宿主对减毒病毒和 wt 病毒的先天性和适应性免疫反应存在重要差异。 HNEC 中的流感病毒复制会诱导鼻粘膜上皮细胞和白细胞亚群内的宿主免疫反应。另一方面，流感病毒编码多种策略，以毒株和细胞类型特异性的方式调节和抑制最初的宿主先天反应，这是流感致病性的重要决定因素。我们假设，测量（a）HNEC以及粘膜和外周淋巴细胞群中对LAIV与wt株的早期宿主转录反应和（b）来自同一个体的外周浆母细胞B细胞以及血清和鼻抗体反应将鉴定特异性预测病毒复制、免疫原性和最终保护的早期免疫标记。我们建议利用 NIH 临床中心正在进行的 A(H1N1)pdm09 流感挑战研究来检验这一假设。我们将从研究对象中收集另外两份鼻拭子，并通过流式细胞术对鼻上皮和淋巴细胞群进行分类，以便随后对粘膜免疫反应和局部病毒复制进行高通量微流控 qRT-PCR 分析。还从同一个体采集血液和鼻洗样本，以测量外周转录反应以及全身 B 细胞和抗体反应。具体目标是： 1) 表征大量未分选的鼻粘膜细胞、分选的 HNEC 和鼻淋巴细胞群以及外周血细胞中的早期宿主免疫转录反应，以确定 wt 病毒诱导的常见和独特的转录特征，并将其与宿主进行比较对 LAIV 免疫的反应，斯坦福大学正在进行的其他研究中正在使用类似的方法进行研究； 2) 在这些相同个体中定量表征外周浆母细胞 B 细胞以及血清和粘膜抗体对 wt 流感感染的反应，并将其与斯坦福大学正在进行的其他 LAIV 研究进行比较，以确定局部和/或系统性早期转录先天免疫反应与随后的浆母细胞和其他适应性局部和全身体液免疫指标相关的标记。这些研究预计将产生关于流感病毒保护性免疫的基本机制的新知识，这将指导改进的季节性和大流行性流感疫苗的开发。