Innate immune response to microbial infection

对微生物感染的先天免疫反应

• 批准号: 8702904
• 负责人: William M. Nauseef
• 金额: $ 182.35万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8702904
• 关键词: Active Immunization; Aerosols; Agonist; Alveolar Macrophages; Anabolism; Animal Model; Antibodies; Antigens; Area; Attention; Bacteria; Biological; Biological Warfare; Biology; Blood capillaries; Breathing; Bulla; Carbohydrates; Categories; Cell Culture Techniques; Cells; Chemicals; Chemistry; Clinical; Complex; Data; Dimensions; Disease; Electron Microscopy; Elements; Endothelial Cells; Endothelium; Endotoxins; Epithelial Cells; Epitopes; Event; Exhibits; Francisella; Francisella tularensis; Funding; Gene Mutation; Genes; Gram-Negative Bacteria; Homologous Gene; Host Defense; Human; Immune; Immune response; Immune system; Immunity; In Vitro; Indium; Infection; Inflammatory Response; Instruction; Invaded; Learning; Left; Libraries; Link; Lipids; Lipopolysaccharides; Lung; Mediating; Medical; Membrane; Microbe; Modeling; Modification; Molecular; Monoclonal Antibodies; Morbidity - disease rate; Mus; Mutagenesis; Natural Immunity; Nature; Organism; Outcome; Passive Immunization; Pathogenesis; Pathogenicity; Phagocytes; Pneumonia; Post-Translational Protein Processing; Production; Program Research Project Grants; Property; Proteins; Public Health; Receptor Cell; Recording of previous events; Recruitment Activity; Regulation; Respiratory Tract Infections; Role; Sepsis; Serogroup B Neisseria meningitidis; Site; Structure; Surface; Testing; Therapeutic; Tularemia; Vaccines; Virulence; Virulence Factors; Work; aerosolized; alveolar epithelium; antimicrobial; base; capillary; capsule; clinical phenotype; cost; cytotoxic; extracellular; genome database; immunogenicity; in vivo; insight; lipooligosaccharide; macrophage; microbial; microorganism; mortality; mutant; neutrophil; novel; pathogen; programs; protective efficacy; protein transport; response; therapeutic development; tool

DESCRIPTION OF THE PPG (provided by applicant): F. tularensis is a potent pathogen when aerosolized, wherein ~ 10 organisms can produce disease. Despite a robust inflammatory response in the lung, mortality from pneumonic tularemia can reach 60%, and the fundamental principles of pulmonary host defense against F. tularensis are incompletely understood. We reason that the remarkable efficiency of aerosolized F. tularensis to produce disease must reflect unique microbial features that render local innate host defenses in the lung quantitatively or qualitatively ineffective. We hypothesize that specific surface features of F. tularensis, especially its extracellular CLM, subvert pulmonary innate defenses, resulting in blunted host response in the lung and consequently the observed virulence of aerosolized F. tularensis. As a corollary, we hypothesize that epitopes on CLM may elicit protective immunity, a concept supported by our data demonstrating that an anti-CLM murine monoclonal antibody exerts protection after passive immunization in an experimental infection challenge. To test our overall hypothesis, we propose an integrated approach to elucidate the properties of F. tularensis and its surface components endotoxin and CLM, and of innate pulmonary host defense that provide the basis for the virulence of inhaled F. tularensis. Accordingly, we have created a program consisting of the following three complementary projects intimately linked to an analytical core: Project 1 - Studies of the capsular antigen of F. tularensis. Project 2 - Early airway innate immune responses to F. tularensis. Project 3 - Human DC and macrophages in dysregulation of lung innate immunity by F. tularensis. The BioAnalytical Chemistry Core will serve as the analytical anchor for structural studies for each of the projects, at critical junctures providing expert analysis of composition, posttranslational modification, or structure of microbial or host proteins, carbohydrates, or lipids. Application of our analytical approaches and the biological principles they support to study interactions of F. tularensis with innate immune elements in the lung, will provide novel and fundamental insights into pathogenicity of F. tularensis disease in the airway and contribute to a better understanding of elements of innate immunity in general. RELEVANCE (See instructions): Understanding how F. tularensis withstands, subverts, or evades innate host defense in the airway will provide important insights into the fundamental biology of F. tularensis and elements of innate host defense in general, and provide potential targets for novel pharmacologic agents and vaccines. Learning how to engage more fully innate immune responses to F. tularensis may have direct therapeutic implications of clinical import, and elucidating the immunogenicity of CLM may provide critical insights into developing an effective vaccine. PROJECT 1: Title: - Studies of the Capsular-like Antigen of F. tularensis Project Leader: Apicella, M PROJECT 1 DESCRIPTION (provided by applicant): F. tularensis is a class A bacterial select agent due to its extreme pathogenicity and potential use as a bioweapon. Early studies by Carlisle and Hood have indicated that F. tularensis produces a capsule-like material (CLM). Many bacteria produce capsules and the genes required to produce them encode proteins for transport, biosynthesis and regulation. Homologs of genes implicated in capsular biosynthesis are present in the Francisella genome database. Preliminary studies, using electron microscopy, indicate and confirm that a CLM surrounds F. tularensis. We can now isolate this material, free from Francisella LPS, based on chemical, chromatographic and immunochemical analysis. This material is loosely associated with the bacterial cell, is easily removed from the bacterial surface and is composed of a repeating tetrasaccharide repeat. Using transposon mutagenesis in F. tularensis Schu S4, we have identified a number of mutants with no or limited reactivity to our CLM specific antibody XE8. We have currently identified the site of insertion of 7 of these transposon mutants that have altered CLM expression that will serve as acapsular strains for the studies in this proposal. Based on these observations, we would pose the following hypotheses 1) F. tularensis expresses a capsular-like material that is important in pathogenesis and that we believe is a group 1 capsule; 2) Alteration of this capsule-like structure by mutations of the genes involved in biosynthesis and expression of CLM will alter pathogenicity of F. tularensis in cell culture models and in an animal model of respiratory infection; 3) Induction of an immune response targeted to the CLM will provide host immunity. The following specific aims will be used to resolve these hypotheses: 1) Characterization of the F. tularensis Schu S4 mutants that are defective in production of CLM, 2) Characterization of the biological role of the F. tularensis Schu S4 CLM, and 3) Study the immunogenicity and protective efficacy of passive and active immunization in murine models of Francisella infection. RELEVANCE (See instructions): The proposed work will focus on characterizing the Francisella CLM which remains poorly understood. There is a significant possibility that efforts to understand this potentially critical virulence factor will not only increase our understanding of Francisella pathogenesis but open up new avenues for therapeutic development, using acapsular strains or antibodies to the CLM.

PPG的描述（由申请人提供）：F。tularensis是一种有效的病原体，当雾化时，约有10种生物可以产生疾病。尽管肺部有强烈的炎症反应，但肺炎的死亡率仍可达到60％，并且肺部宿主防御针对F. tolarensis的基本原理尚不完全了解。我们认为，雾化杆菌对产生疾病的显着效率必须反映出独特的微生物特征，从而在定量或定性上无效地使肺部宿主防御能力。我们假设F. toarlensis的特定表面特征，尤其是其细胞外CLM，颠覆了肺的先天防御，从而导致肺中的宿主反应钝化，因此观察到了雾化的F. toarlensis的毒力。作为推论，我们假设对CLM的表位可能会引起保护性免疫，我们的数据支持了一个概念，表明一种抗CLM鼠单克隆抗体在实验感染挑战中被动免疫后具有保护。为了检验我们的总体假设，我们提出了一种综合方法，以阐明tularensis及其表面成分内毒素和CLM的性质，以及先天肺宿主防御的特性，为吸入f。tularensis毒力提供基础。因此，我们创建了一个计划，该程序由以下三个互补项目组成，与分析核心密切相关：项目1-对tulareensis的囊囊抗原的研究。项目2-早期气道先天性免疫反应对f。tularensis。项目3- F. tularensis对肺部先天免疫失调的人类直流和巨噬细胞。生物分析化学核心将作为每个项目的结构研究的分析锚，在关键时刻提供了对组成，翻译后修饰或微生物或宿主蛋白，碳水化合物或脂质的结构的专家分析。我们的分析方法的应用及其支持的生物学原理在研究肺部与先天免疫元素的相互作用中的相互作用将提供对气道中F. tolarensis疾病的致病性的新颖和基本见解，并有助于更好地了解一般性免疫的元素。 相关性（请参阅说明）：了解F. tularensis在气道中如何承受，颠覆或逃避先天的宿主防御能力将为F. tularensis的基本生物学以及一般性宿主防御的基本生物学提供重要的见解，并提供一般的新型药理药物和疫苗的潜在目标。学习如何使对f的F. tularensis的更完全先天的免疫反应可能具有临床进口的直接治疗意义，并阐明CLM的免疫原性可能会为开发有效的疫苗提供关键的见解。 项目1： 标题： - tularensis的囊状抗原的研究 项目负责人：Apicella，M 项目1描述（由申请人提供）：F。tularensis是A类细菌选择剂，因为其极端的致病性和潜在用作生物武器。 Carlisle和Hood的早期研究表明，F。tularensis产生类似胶囊的材料（CLM）。许多细菌产生胶囊，产生它们所需的基因编码蛋白质以进行运输，生物合成和调节。弗朗西斯菌基因组数据库中存在与囊囊生物合成有关的基因的同源物。使用电子显微镜的初步研究表明并确认CLM围绕F. tularensis。现在，我们可以根据化学，色谱和免疫化学分析分离这种材料，而不受Francisella LPS的隔离。该材料与细菌细胞松散相关，很容易从细菌表面去除，并由重复的四糖重复组成。使用F. tularensis Schu S4中的转座子诱变，我们已经确定了许多对CLM特异性抗体XE8反应性无反应性的突变体。我们目前已经确定了这些转座子突变体中的7个插入部位，这些突变体改变了CLM表达，这将作为该提案中研究的交叉菌株。基于这些观察结果，我们将提出以下假设1）F。tularensis表达在发病机理中很重要的囊状材料，我们认为这是1组胶囊； 2）通过在细胞培养模型和呼吸道感染的动物模型中，参与生物合成和CLM表达的基因的突变改变了这种胶囊样结构； 3）诱导针对CLM的免疫反应将提供宿主免疫。 The following specific aims will be used to resolve these hypotheses: 1) Characterization of the F. tularensis Schu S4 mutants that are defective in production of CLM, 2) Characterization of the biological role of the F. tularensis Schu S4 CLM, and 3) Study the immunogenicity and protective efficacy of passive and active immunization in murine models of Francisella infection. 相关性（请参阅说明）：拟议的工作将集中于表征弗朗西斯氏菌CLM，该CLM的理解仍然很差。通过使用与CLM的囊性菌株或抗体，了解这种潜在的关键毒力因子的努力不仅会增加我们对Francisella发病机理的理解，而且还可以为治疗发育开辟新的途径。

项目成果

Editorial: Gazing forward while looking back.

社论：回顾过去，凝视前方。

• DOI: 10.1189/jlb.0812424
• 发表时间: 2013
• 期刊: Journal of leukocyte biology
• 影响因子: 5.5
• 作者: Cassatella,MarcoA;Nauseef,WilliamM
• 通讯作者: Nauseef,WilliamM

Coordinate regulation of sphingosine kinase and actin dynamics.

鞘氨醇激酶和肌动蛋白动力学的协调调节。

• DOI: 10.1007/978-1-59745-396-7_21
• 发表时间: 2009
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Iyer,ShankarS;Kusner,DavidJ
• 通讯作者: Kusner,DavidJ

Pondering neutrophil extracellular traps with healthy skepticism.

• DOI: 10.1111/cmi.12652
• 发表时间: 2016-10
• 期刊: Cellular microbiology
• 影响因子: 3.4
• 作者: Nauseef WM;Kubes P
• 通讯作者: Kubes P

The role of complement opsonization in interactions between F. tularensis subsp. novicida and human neutrophils.

• DOI: 10.1016/j.micinf.2009.04.016
• 发表时间: 2009-07
• 期刊: Microbes and infection
• 影响因子: 5.8
• 作者: Barker JH;McCaffrey RL;Baman NK;Allen LA;Weiss JP;Nauseef WM
• 通讯作者: Nauseef WM

Neutrophils, from cradle to grave and beyond.

• DOI: 10.1111/imr.12463
• 发表时间: 2016-09
• 期刊: Immunological reviews
• 影响因子: 8.7
• 作者: Nauseef WM