Pneumocystis carinii: Macrophage & Epithelial Activation

卡氏肺孢子虫：巨噬细胞

• 批准号: 10320933
• 负责人: ANDREW H LIMPER
• 金额: $ 39.75万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-07-10 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10320933
• 关键词: Acquired Immunodeficiency Syndrome; Adaptor Signaling Protein; Address; Adoptive Transfer; Alveolar Macrophages; Animals; Antibiotic Therapy; Antibiotics; Automobile Driving; C Type Lectin Receptors; CD4 Positive T Lymphocytes; Caspase; Cell Therapy; Cell Wall; Cell surface; Cells; Cessation of life; Characteristics; Complication; Data; Dendritic Cells; Development; Epithelial; Epithelial Cells; Equilibrium; Europe; Event; Exhibits; Family; Feasibility Studies; Future; Host Defense; Human; Human Characteristics; Immune; Immune response; Immunity; Immunocompetent; Impairment; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Kinetics; Lentivirus; Lung; Lymphocyte; Mediating; Mitogen-Activated Protein Kinases; Molecular; Mus; Natural Immunity; Organism; PC3 cell line; Patients; Phenotype; Pneumocystis; Pneumocystis carinii; Pneumocystis carinii Pneumonia; Pulmonary Inflammation; Receptor Signaling; Refractory; Resource-limited setting; Respiratory Failure; Respiratory Insufficiency; Rodent; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Sodium Glutamate; Survival Rate; System; Testing; Therapeutic Uses; Time; Wild Type Mouse; alveolar epithelium; base; beta-Glucans; cytokine; dectin 1; immunosuppressed; improved; lung injury; macrophage; monocyte; mortality; mouse model; novel strategies; novel therapeutics; overexpression; peripheral blood; pre-clinical; receptor function; recruit; respiratory; response; treatment strategy; uptake

ABSTRACT: Pneumocystis pneumonia (PCP) remains a serious complication of AIDS.1-5 Pc thrives during impaired lymphocytic immunity, leading to respiratory failure.3,6-8 To better understand host responses during CD4 deficiency, we studied host innate immunity in PCP, demonstrating central roles for alveolar macrophages (AMS) in Pc clearance.9 We also defined activity of Pc associated inflammation from AMS and alveolar epithelial cells (AECs) leading to respiratory impairment.10,11 Interactions of Pc β-glucans with the C-Type Lectin Receptor (CLR) Dectin-1 and interactions of Pc MSG with the CLR Mincle initiate clearance of Pc, but also trigger damaging lung inflammation.4,12-16 Understanding mechanisms to restore balance between Pc clearance and inflammation holds potential benefit for treating severe PCP. Since the last submission, we have compelling new data demonstrating central roles across the CLR family during PCP both in CD4 depleted and in immunocompetent hosts using CARD9-/- mice. CARD9, a Caspase Recruitment Domain (CARD) adaptor protein, is the central molecule through which Dectin-1, Mincle, and other CLRs signal.17 Hence, CARD9-/- mice exhibit a deficiency of most CLR immunity. Notably, CD4-deficient CARD9-/- mice had impaired killing of Pc, with markedly greater Pc burdens and concurrently a dramatic reduction of inflammation compared to wild-type mice. Immune competent CARD9-/- mice also had significantly impaired Pc clearance. Despite this phenotype, the molecular events mediated by CARD9 during PCP are unknown. We hypothesize that impairment of CARD9, a central regulator of major CLR function, leads to broad deficiency critical for Pc clearance and inflammatory responses. This will be addressed through three independent but interrelated Aims. In Aim 1, we will evaluate the role of CARD9 in driving Pc clearance and inflammation following interactions with Pc. We will study uptake and killing of Pc, cytokine release, polarization of macrophages, and the cell signaling mechanisms in CARD9-/- versus wild type AMS, AECs and dendritic cells. Next, in Aim 2, we will determine the impact of CARD9 on Pc clearance, inflammation, and mortality in immune competent and CD4 deficient CARD9-/- mice during PCP. We will examine the major causes of lethality during PCP, namely Pc burden and lung inflammation by performing kinetic studies of Pc-infected CARD9-/- mice using both CD4 depleted and immunocompetent mice. Finally, in Aim 3, we will explore cell based treatments targeting CARD9 as adjunctive therapies for PCP. While cell based treatment strategies are not needed for mild to moderate PCP, such therapies may be beneficial in addition to antibiotics in severe PCP with mortality over 80%.5,18 We have demonstrated that transfer of M2 polarized macrophages during PCP signficantly enhances Pc clearance and improves lung inflammation.19 Therefore, we will test the utlility of macrophages driven to overexpress CARD9 and CLR as adjuctive therapy during severe PCP. These studies will define the activity of the important CLR/CARD9 system in host defense during PCP, which may be exploited for novel therapies for Pc infection.

摘要：肺炎胸膜肺炎（PCP）仍然是辅助的严重并发症。1-5PC在 淋巴细胞免疫学受损，导致呼吸衰竭。3,6-8为了更好地理解宿主的反应 CD4缺乏症，我们研究了PCP中的固有免疫力，证明了肺泡巨噬细胞的核心作用 （AMS）在PC间隙中。9我们还定义了PC相关的AM和肺泡的炎症的活性 上皮细胞（AEC）导致呼吸障碍。10,11PCβ-葡聚糖与C型的相互作用 凝集素受体（CLR）Dectin-1和PC MSG与CLR Mincle的相互作用启动PC的清除率，但 还会触发破坏肺部注射。4,12-16理解机制以恢复PC之间的平衡 清除和炎症具有治疗严重PCP的潜在好处。自上次提交以来，我们有 引人入胜的新数据，证明了CD4中PCP期间CLR家族的主要角色耗尽和 在使用Card9 - / - 小鼠的免疫能力宿主中。 Card9，caspase招聘域（卡）适配器 蛋白质是中央分子，dectin-1，mincle和其他CLR信号。17因此，card9 - / - 小鼠 呼出大多数CLR免疫力的不足。值得注意的是，CD4缺陷卡9 - / - 小鼠杀死PC的损害， 与野生型相比 老鼠。免疫胜任的Card9 - / - 小鼠的PC清除率也大大受损。尽管这种表型， 在PCP期间由Card9介导的分子事件尚不清楚。我们假设 Card9是主要CLR功能的中央调节器，导致广泛的缺陷对于PC清除至关重要 和炎症反应。这将通过三个独立但相互关联的目标解决。目标 1，我们将评估与PC相互作用后，Card9在驾驶PC清除率和炎症中的作用。 我们将研究PC的摄取和杀伤，细胞因子释放，巨噬细胞的极化以及细胞信号传导 Card9 - / - 与野生型AM，AEC和树突状细胞中的机制。接下来，在AIM 2中，我们将确定 Card9对免疫能力和CD4不足的PC清除，感染和死亡率的影响 pcp期间的card9 - / - 鼠标。我们将检查PCP期间致死性的主要原因，即PC Burnen和 通过使用CD4耗尽和 免疫能力小鼠。最后，在AIM 3中，我们将探索针对Card9的基于细胞的治疗方法 PCP的辅助疗法。轻度至中度PCP不需要基于细胞的治疗策略，但 此类疗法除了严重PCP的抗生素外，死亡率超过80％。5,18我们有有益 证明PCP期间M2极化巨噬细胞的转移显着增强了PC清除率和 改善肺部注射。19因此，我们将测试驱动到过表达Card9的巨噬细胞的UTLITY 在严重的PCP期间，CLR作为添加剂疗法。这些研究将定义重要的活动 PCP期间宿主防御中的CLR/CARD9系统，可以利用用于PC感染的新型疗法。