Regulatory Tcells and the fibroproliferative response in Acute Lung Injury

调节性 T 细胞和急性肺损伤中的纤维增殖反应

• 批准号: 8136644
• 负责人: Brian Garibaldi
• 金额: $ 5.12万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8136644
• 关键词: Accounting; Acute Lung Injury; Adoptive Transfer; Adult Respiratory Distress Syndrome; Affect; Animal Model; Bleomycin; Bone Marrow; Cell Count; Cell Culture Techniques; Cells; Coculture Techniques; Collagen; Conditioned Culture Media; Data; Deposition; Development; Diffuse; Disease; Edema; Environmental air flow; Enzyme-Linked Immunosorbent Assay; Extracellular Matrix; Extracellular Matrix Proteins; Fibroblasts; Fibronectins; Fibrosis; Fluorescence-Activated Cell Sorting; Future; Goals; Histology; Hour; Human; Hyaluronan; IL2RA gene; Immunoblotting; Immunohistochemistry; In Vitro; Individual; Infiltration; Inflammation; Inflammatory; Injury; Laminin; Lead; Liquid substance; Lung; Lung Compliance; Lung diseases; Lymphocyte; Macrophage Activation; Measures; Membrane; Metalloproteases; Modeling; Morbidity - disease rate; Mus; Myofibroblast; Outcome; Pathogenesis; Patients; Phase; Phenotype; Play; Proteins; Rag1 Mouse; Recruitment Activity; Regulatory T-Lymphocyte; Research; Resolution; Role; Sampling; Smooth Muscle Actin Staining Method; Staging; Staining method; Stains; Stem cells; Stimulus; Techniques; Tidal Volume; Tissues; United States; arm; epithelial to mesenchymal transition; improved; in vivo; inhibitor/antagonist; injured; interest; lung injury; macrophage; mortality; mouse model; novel; novel therapeutics; public health relevance; research study; response; Accounting; Acute Lung Injury; Adoptive Transfer; Adult Respiratory Distress Syndrome; Affect; Animal Model; Bleomycin; Bone Marrow; Cell Count; Cell Culture Techniques; Cells; Coculture Techniques; Collagen; Conditioned Culture Media; Data; Deposition; Development; Diffuse; Disease; Edema; Environmental air flow; Enzyme-Linked Immunosorbent Assay; Extracellular Matrix; Extracellular Matrix Proteins; Fibroblasts; Fibronectins; Fibrosis; Fluorescence-Activated Cell Sorting; Future; Goals; Histology; Hour; Human; Hyaluronan; IL2RA gene; Immunoblotting; Immunohistochemistry; In Vitro; Individual; Infiltration; Inflammation; Inflammatory; Injury; Laminin; Lead; Liquid substance; Lung; Lung Compliance; Lung diseases; Lymphocyte; Macrophage Activation; Measures; Membrane; Metalloproteases; Modeling; Morbidity - disease rate; Mus; Myofibroblast; Outcome; Pathogenesis; Patients; Phase; Phenotype; Play; Proteins; Rag1 Mouse; Recruitment Activity; Regulatory T-Lymphocyte; Research; Resolution; Role; Sampling; Smooth Muscle Actin Staining Method; Staging; Staining method; Stains; Stem cells; Stimulus; Techniques; Tidal Volume; Tissues; United States; arm; epithelial to mesenchymal transition; improved; in vivo; inhibitor/antagonist; injured; interest; lung injury; macrophage; mortality; mouse model; novel; novel therapeutics; public health relevance; research study; response

DESCRIPTION (provided by applicant): Acute lung Injury (ALI) and the more severe Acute Respiratory Distress Syndrome (ARDS) are diffuse lung diseases that cause tremendous morbidity and mortality in the United States each year. Despite decades of research into the pathogenesis and mechanisms underlying the development of ARDS, mortality remains high and the only treatment shown to be effective is low tidal volume ventilation in patients who require ventilatory support. A subset of patients with ARDS progress to a fibroproliferative stage marked pathologically by fibrosis and clinically by increasing dead space, decreasing lung compliance and worsening oxygenation. Recent interest has shifted from the factors that promote the development of ALI to determinants of the resolution of lung injury. Regulatory T cells (Tregs) have been found to be critical to the resolution of ALI in a mouse model of LPS-induced lung injury. They have also been shown to increase in number in the BAL fluid of patients with ARDS, suggesting that they likely play an important role in the normal human response to ALI. The role of Tregs in the fibroproliferative response to ALI has not been studied. The goal of this proposal is to determine the role of Tregs in the fibroproliferative response to ALI, with the hope of identifying novel mechanisms that may ultimately lead to future treatment options in patients with ALI. In order to achieve this goal this proposal will utilize both in vivo and in vitro techniques to examine the role of Tregs in fibroproliferation after LPS-induced ALI. For the in vivo arm, C57BL/6 wildtype (WT) mice and Rag-1-/- lymphocyte deficient mice will be exposed to intratracheal (i.t.) LPS and the resulting fibroproliferative response will be rigorously phenotyped. Adoptive transfer of Tregs into Rag-1-/- mice will be employed to examine the specific effects of Tregs on the fibroproliferative response. In the in vitro experiments, fibroblasts will be grown in cell culture under various stimulating conditions. Tregs and macrophages will be added in different co-culture combinations to examine the cellular mechanisms by which Tregs modify fibroblast function. These experiments will explore new observations on the role of Tregs in the fibroproliferative response to ALI with the ultimate goal of improving patient outcomes in this oftentimes fatal disease. PUBLIC HEALTH RELEVANCE: Acute lung injury (ALI) is a common and oftentimes fatal pulmonary disease that may result in lung fibrosis. T- regulatory (Treg) cells have been shown to be important in the resolution of ALI but their role in the development of fibrosis after ALI is unknown. We propose to study the effect of Tregs on the fibroproliferative response to ALI with the hope of identifying novel mechanisms that may ultimately lead to new therapeutic options for this disease.

描述（由申请人提供）：急性肺损伤（ALI）和更严重的急性呼吸窘迫综合征（ARDS）是每年在美国引起巨大发病率和死亡率的弥漫性肺部疾病。尽管数十年来研究了ARDS发展的发病机理和机制，但死亡率仍然很高，并且证明有效的唯一治疗方法是需要通气支持的患者潮汐量低。 ARDS患者的一部分通过纤维化和临床上的纤维增生性阶段而进展，通过增加死亡空间，降低肺合顺速性并恶化氧合。最近的兴趣已从促进ALI发展的因素转变为肺损伤解决的决定因素。已发现调节性T细胞（TREG）对于在LPS诱导的肺损伤的小鼠模型中对ALI的分辨率至关重要。还显示它们在ARDS患者的BAL液中的数量增加，这表明它们可能在人类对ALI的正常反应中起重要作用。尚未研究Treg在对ALI的纤维增生反应中的作用。该提案的目的是确定Treg在对ALI的纤维增生性反应中的作用，希望确定新的机制，这些机制最终可能导致ALI患者的未来治疗选择。为了实现这一目标，该提案将利用体内和体外技术来检查Tregs在LPS诱导的ALI后纤维增生中的作用。对于体内组，C57BL/6野生型（WT）小鼠和RAG-1 - / - 淋巴细胞缺陷小鼠将暴露于气管内（I.T.）LPS，并将其结果纤维增生性反应进行严格的表型。将Tregs的收养转移到RAG-1 - / - 小鼠中，将用于检查Tregs对纤维增生反应的特定影响。在体外实验中，在各种刺激条件下，成纤维细胞将在细胞培养中生长。 Treg和巨噬细胞将以不同的共培养组合添加，以检查Tregs修改成纤维细胞功能的细胞机制。这些实验将探讨有关Treg在对ALI纤维增生反应中作用的新观察结果，其最终目的是改善这种致命疾病的患者结局。 公共卫生相关性：急性肺损伤（ALI）是一种常见的，通常是致命的肺疾病，可能导致肺纤维化。 T-调节（Treg）细胞已被证明在ALI的分辨率中很重要，但是它们在ALI后其在纤维化发展中的作用尚不清楚。我们建议研究Treg对ALI纤维增生性反应的影响，以期识别新的机制，这些机制最终可能导致该疾病的新治疗选择。