Innate inflammatory responses during pulmonary infections

肺部感染期间的先天炎症反应

• 批准号: 10272218
• 负责人: Katrin Mayer-Barber
• 金额: $ 129.37万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10272218
• 关键词: Address; Adipose tissue; Aerosols; Animal Model; Animals; Antibodies; Area; Asthma; Autoimmune Diseases; Bacterial Infections; Biological; Biological Process; Blood specimen; Bone Marrow; Cations; Cell Communication; Cell Death; Cells; Chinese People; Chronic; Chronic Disease; Clinical Research; Cryptococcus; Cytoplasmic Granules; Data; Development; Dinoprostone; Disease; Dose; Effector Cell; Eicosanoids; Eotaxin; Exhibits; Fibroblasts; Fibrosis; Flow Cytometry; Goals; Helminths; Host resistance; Human; Hypersensitivity; IL8 gene; Immune response; Immunity; Immunologic Techniques; Immunologics; Immunotherapy; Individual; Infection; Inflammation; Inflammatory; Inflammatory Response; Influenza A virus; Innate Immune Response; Interferon Type I; Interferons; Interleukin-1; Interleukin-6; Investigation; Lead; Leukotrienes; Light; Lipids; Lung; Lung infections; Macaca; Macaca mulatta; Maintenance; Malignant Neoplasms; Measures; Mediating; Messenger RNA; Modeling; Mouse Strains; Mus; Mycobacterium avium; Mycobacterium tuberculosis; Mycoses; Myelogenous; Nature; Outcome; Pathology; Pathway interactions; Patients; Phagocytes; Physiological; Plasma Cells; Play; Predisposition; Primary Infection; Process; Prospective cohort; Prostaglandins; Proteins; Published Comment; Reactive Oxygen Species; Regulation; Research; Resected; Respiratory Tract Infections; Role; Stains; Surface; System; TNF gene; Time; Tissues; Tuberculosis; Vaccine Design; Vaccines; Viral; Whole Blood; adaptive immunity; antimicrobial; arm; autoinflammatory; bactericide; chemokine; chronic infection; cytokine; cytotoxic; eosinophil; granulocyte; immunoregulation; in vivo; insight; lipid mediator; macrophage; migration; mouse model; neutrophil; non-tuberculosis mycobacteria; novel; programs; respiratory; response; translational pipeline; treatment strategy; wound healing

The development of effective vaccines and host directed therapies (HDT) against M. tuberculosis (Mtb) infection requires a detailed understanding of the cellular basis of protective immunity. Considerable progress has been made in our understanding of protective adaptive immunity, yet relatively little is known about the contribution of innate effector cells. Particularly, the biological relevance of granulocytes like neutrophils and eosinophils is poorly understood. The innate inflammatory response is a prime target for HDT and manipulation of granulocytes could have major inflammatory and immunoregulatory implications for host resistance. Recent advances in vaccine design were highlighted by our group through an invited commentary in Immunity. Similar to neutrophils, eosinophils are phagocytic cells of the myeloid lineage that are thought to play an important effector role in the innate immune response. Their lineage-specific secondary granules contain cytotoxic cationic granular proteins (ECP, EDN, MBP and EPO) that have been shown to exhibit anti-microbial activity and cause tissue damage. Preformed cytokines contained in these same granules include inflammatory cytokines like TNF-, IL-1, IL-6 and IL-8 and pro-fibrogenic cytokines that can stimulate fibroblast proliferation, fibrotic and wound healing responses. In addition, lipid bodies, which form in response to eosinophil activation, contain a wide variety of leukotrienes, prostaglandins and reactive oxygen species that can contribute to these processes. More recently, eosinophils have been shown to play an important role in immunoregulation and homeostatic functions, including maintenance of long-lived plasma cells in the bone marrow and alternatively activated macrophages in adipose tissue. Eosinophils and their biological functions of have been studied primarily in the context of type 2 immunity, including parasitic helminth and pulmonary fungal infection, allergies and asthma. The role of eosinophils during bacterial infection remains largely unexplored. However, a comprehensive study on the role of eosinophils during Mtb infection is lacking, both in animal models as well human clinical studies. Another barrier to our understanding of eosinophils in host resistance to TB is a paucity of data from the mouse model of Mtb infection. There are likely two major contributing factors to be considered for this: 1) Mtb infection primarily causes a type I immune response, and eosinophils are rare in numbers compared to neutrophils in the lungs of Mtb infected mice (KDMB unpublished data) and 2) the pathology of Mtb infected mouse lungs and human lungs is vastly different. Eosinophils were found to be enriched in areas of tissue remodeling and fibrosis in TB resected lungs from patients (KDMB unpublished data). While fibrotic responses are often seen in TB patients, they are not considered a feature in the mouse model of Mtb infection. Indeed, the physiological role and function of eosinophils in type 1 immune responses to intracellular bacterial infections remains unknown. Here, in exploring the pulmonary granulocytic response to Mtb infection across species, we discovered that eosinophils migrated rapidly to Mtb-infected lungs. Because surface antibodies used to identify human eosinophils did not cross-react with eosinophils from rhesus macaques, we developed a strategy to quantify eosinophil responses in macaques by flow cytometry. We found that intracellular staining with EPX selectively and specifically stains eosinophils in both NHP and human whole blood samples and demonstrated that eosinophils rapidly migrate into Mtb infected rhesus macaque airways. We also identified eosinophils as the first cells sensing and repsondong to Mtb infection in the mouse lung after Mtb infection. Importantly, eosinophil deficient mouse strains demonstrated increased susceptibility to disease, with significantly reduced survival times. We also examined the chemokines responsible for eosinophil migration during tuberculosis and found that CCR3-mediated recognition of eotaxins were dispensible for tissue migration. We have previously characterized two major innate cytokine pathways, IL-1 and type I interferons, respectively, that play pivotal roles in governing host resistance versus disease in the murine model of Mtb infection by intersecting the eicosanoid lipid network. In particular, we uncovered that IL-1 can in turn counter-regulate type I IFN driven detrimental responses during Mtb infection. In murine and human macrophages IL-1 and IL-1 potently inhibit type I IFN induction at both the mRNA and protein level and similarly IFN mRNA and protein levels are upregulated in the lungs of Mtb infected Il1r1-/- deficient mice. This inhibition is of functional importance because mice doubly deficient in Il1r1,Ifnar1-/- are partially protected while Il1r1-/- singly deficient animals succumb rapidly to Mtb aerosol challenge. Moreover, when IL-1 is present in type I IFN treated cultures, it even suppresses the pro-bacterial effects downstream of IFN that lead to increased bacterial replication. Interestingly, IL-1 induced PGE2 is also able to potently inhibit type I IFNs in a dose dependent manner. These data highlighted and provided proof-of-concept that the cross-talk of IL-1 and type I IFN provides a valuable target for host-directed therapies of Mtb and plays a major role during infection in mice. Moreover, we measured and analyzed cytokines, chemokines, and lipid mediators of the IL-1, type IFN and eicosanoid axis in a prospective cohort of Chinese individuals infected with M. tuberculosis. We are currently investigating the exact nature of IL-1 and type I IFN induced cell death mechanisms, lipid regulation and host resistance pathways during tuberculosis.

针对结核分枝杆菌（MTB）感染的有效疫苗和宿主定向疗法（HDT）的开发需要详细了解保护性免疫的细胞基础。我们对保护性适应性免疫的理解已经取得了长足的进步，但对先天效应细胞的贡献知之甚少。尤其是，对中性粒细胞和嗜酸性粒细胞等粒细胞的生物学相关性知之甚少。先天炎症反应是HDT的主要靶标，粒细胞的操纵可能对宿主耐药性具有重大的炎症和免疫调节意义。疫苗设计的最新进展是通过邀请的免疫评论来强调的。 与中性粒细胞相似，嗜酸性粒细胞是髓样谱系的吞噬细胞，被认为在先天免疫反应中起重要作用。它们的谱系特异性二级颗粒含有细胞毒性阳离子颗粒蛋白（ECP，EDN，MBP和EPO），已显示出表现出抗微生物活性并引起组织损伤。这些相同颗粒中包含的预先形成的细胞因子包括炎症细胞因子，例如TNF-，IL-1，IL-6和IL-8以及促纤维化细胞因子，可以刺激成纤维细胞增殖，纤维化和伤口愈合反应。此外，响应嗜酸性粒细胞激活形成的脂质体包含各种各样的白三烯，前列腺素和活性氧，可以有助于这些过程。最近，嗜酸性粒细胞已被证明在免疫调节和稳态功能中起着重要作用，包括维持骨髓中长寿命的浆细胞和脂肪组织中的巨噬细胞。嗜酸性粒细胞及其生物学功能主要是在2型免疫力的背景下进行的，包括寄生虫蠕虫和肺真菌感染，过敏和哮喘。嗜酸性粒细胞在细菌感染中的作用在很大程度上尚未开发。然而，在动物模型以及人类的临床研究中，都缺乏关于嗜酸性粒细胞在MTB感染中的作用的全面研究。我们对宿主对结核病抗性中嗜酸性粒细胞的理解的另一个障碍是MTB感染小鼠模型的数据很少。为此，可能要考虑的主要因素有两个主要因素：1）MTB感染主要引起I型免疫反应，而与MTB感染小鼠的肺中性粒细胞相比，嗜酸性粒细胞的数量很少，而MTB感染小鼠（KDMB未发表的数据）和2）MTB感染小鼠Lungs和人类Lungs的病理学很大。发现嗜酸性粒细胞富集在患者的结核病（KDMB未发表数据）的结核病中组织重塑和纤维化区域。尽管在结核病患者中经常看到纤维化反应，但在MTB感染的小鼠模型中，它们并未被认为是它们的特征。实际上，嗜酸性粒细胞在1型对细胞内细菌感染的免疫反应中的生理作用和功能仍然未知。在这里，在探索跨物种对MTB感染的肺粒细胞反应时，我们发现嗜酸性粒细胞迅速迁移到MTB感染的肺部。由于用于鉴定人嗜酸性粒细胞的表面抗体与猕猴的嗜酸性粒细胞没有交叉反应，因此我们制定了一种通过流式细胞仪中猕猴定量嗜酸性粒细胞反应的策略。我们发现，在NHP和人类全血样品中选择性地，特异性地对EPX进行了细胞内染色，并证明嗜酸性粒细胞迅速迁移到感染了MTB的恒河猴猕猴气道中。我们还确定嗜酸性粒细胞是MTB感染后小鼠肺中MTB感染的第一个细胞感应的细胞。重要的是，嗜酸性粒细胞缺乏小鼠菌株表明对疾病的敏感性增加，生存时间大大降低。我们还检查了结核病期间负责嗜酸性粒细胞迁移的趋化因子，发现CCR3介导的eotaxins识别可用于组织迁移。 我们先前曾分别表征了两种主要的先天细胞因子途径IL-1和I型干扰素，它们通过与类eicosanoid脂质网络相结合，在MTB感染的鼠模型中扮演着宿主抗性与疾病的关键作用。特别是，我们发现IL-1反过来又可以反调节I型IFN在MTB感染过程中造成的有害反应。在鼠和人类巨噬细胞中，IL-1和IL-1在mRNA和蛋白质水平上有效抑制I型IFN诱导，并且类似地，IFN mRNA和蛋白质水平在感染IL1R1/ - 缺陷的MTB肺中上调。这种抑制作用具有功能重要性，因为小鼠在IL1R1中偶尔缺乏IFNAR1 - / - 受到部分保护，而IL1R1 - / - 单一单一缺陷的动物迅速屈服于MTB气溶胶挑战。此外，当I型IFN治疗培养物中存在IL-1时，它甚至会抑制IFN下游的促细菌作用，从而导致细菌复制增加。有趣的是，IL-1诱导的PGE2也能够以剂量依赖性的方式有效抑制I型IFN。这些数据强调并提供了概念验证，即IL-1和I型IFN的串扰为MTB的宿主指导疗法提供了宝贵的目标，并在小鼠感染中起着重要作用。 此外，我们在感染了结核分枝杆菌的中国个体中，测量和分析了IL-1，IFN和类花生烷轴的细胞因子，趋化因子和脂质介质。我们目前正在研究结核病期间IL-1和I型IFN诱导的细胞死亡机制，脂质调节和宿主抗性途径。