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（由申请方提供）：2009年由A（H1N1）pdm 09病毒引起的流感（流感）大流行和高致病性禽流感A病毒感染的零星人间病例强调了对季节性和潜在大流行性流感毒株的改良流感疫苗的需求。流感疫苗接种的一种方法需要鼻内施用减毒活流感病毒（LAIV），其部分模拟病毒感染上呼吸道的人鼻上皮细胞（HNEC）中的天然途径和位点。然而，与天然野生型（wt）流感感染相比，LAIV未能诱导强的全身免疫应答，表明宿主对减毒病毒和wt病毒的先天性和适应性免疫应答的重要差异。HNEC中的流感病毒复制导致在鼻粘膜的上皮细胞和白细胞细胞亚群内诱导宿主免疫应答。另一方面，流感病毒编码多种策略来以毒株特异性和细胞类型特异性的方式调节和抑制初始宿主先天应答，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体中对LAIV vs. wt毒株的早期宿主转录应答和（B）来自相同个体的外周浆母细胞B细胞和血清及鼻抗体应答将鉴定预测病毒复制、免疫原性和最终保护的特异性早期免疫标志物。我们建议利用NIH临床中心正在进行的甲型H1N1流感pdm 09挑战研究来验证这一假设。我们将从研究受试者中收集另外两个鼻拭子，并通过流式细胞术对鼻上皮细胞和淋巴细胞群进行分类，用于随后的粘膜免疫应答和局部病毒复制的高通量微流控qRT-PCR分析。还从相同的个体收集血液和鼻洗液样品，以测量外周转录应答和全身B细胞和抗体应答。具体目标是：1）表征在大量未分选的鼻粘膜细胞和分选的HNEC和鼻淋巴细胞群体中以及在外周血细胞中的早期宿主免疫转录应答，以鉴定由wt病毒诱导的共同和独特的转录特征，并将它们与对LAIV免疫的宿主应答进行比较，这在斯坦福大学的其他正在进行的研究中正在用类似的方法进行研究;和2）在这些相同的个体中以定量水平表征外周浆母细胞B细胞和血清以及粘膜抗体对野生型流感感染的应答，并将这些与斯坦福大学正在进行的LAIV研究进行比较，鉴定与随后的浆母细胞和其它适应性局部和全身体液免疫指标相关的局部和/或全身早期转录先天免疫应答标志物。预计这些研究将产生关于流感病毒保护性免疫的潜在机制的新知识，这将指导改进的季节性和大流行性流感疫苗的开发。