CC Chemokines in Allergic Asthma

过敏性哮喘中的 CC 趋化因子

• 批准号: 6908970
• 负责人: C Edward Rose
• 金额: $ 33.3万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6908970
• 关键词: active immunization; asthma; cell migration; chemokine; chemokine receptor; cytokine; dendritic cells; flow cytometry; helper T lymphocyte; laboratory mouse; monocyte chemoattractant protein 1; neutralizing antibody; passive immunization; pathologic process; respiratory hypersensitivity

DESCRIPTION (provided by applicant): Asthma is a major public health issue, and its pathogenesis is complex. In the particular, the role of CC chemokines in this disease is poorly understood. We have successfully developed a new animal model in which antigen (Ag)-pulsed dendritic cells (DC) are able to induce airway hyperreactivity (AHR), goblet cell hyperplasia, specific IgE response, hypereosinophilia, and marked lung inflammation. We have also shown that in mutant mice deficient in CC chemokine receptor 2 (CCR2-/-), an accelerated Th2 response is seen in the lung after aeroallergen challenge. Others and we have shown that CCR2 deficiency alters the migration of monocytes, and possibly other cells, such as Th lymphocytes. In this proposal, the role of MCP-1 in the induction of various pathological changes will be investigated. In addition, altered leukocyte migration, as related to the pathogenesis of asthma, will be studied. Three specific aims are proposed. Aim 1: To demonstrate that elevated MCP- 1 plays a major role in the observed accentuated Th2 response in the lung in the Ag induced asthma model, using either neutralizing anti-MCP-1 antibody 2H5 or CCR2-/-/MCP-/--1 double mutant mice. Aim 2: To determine the kinetics of lymphocyte, monocyte and eosinophil migration to the lungs and airways in allergen-challenged CCR2-/- mice. This will be done by studying mice at increasing intervals after onset of Ag-challenge using flow cytometry on total lung leukocytes, or immunohistochemical staining of lung tissue. Antibodies to characterization leukocytes will include F4/80 (macrophage), anti-CD4 mAb, anti-CD8 mAb, Thi (IFNghigh, IL-5low), Th2 (IFNglow, IL-5high), anti-MHC class I mAb (antigen presenting cells), and anti-DEC-205 (DC). Aim 2 will also evaluate lung trafficking of adoptively transferred Tg+ CD4 cells from CCR2-/- or CCR2+/+ DO 11.10 mice, to address the issue of whether altered numbers of Th subsets in the lung are related to trafficking or in situ differentiation. Aim 3: To determine the relative role of monocytes and lymphocytes in the accentuated Th2 response in the allergen-challenged CCR2-/- mice. This will be done using chimeric mice created by adoptive transfer of CCR2+/+ or CCR2-/- Tg+ DO 11.10 CD4 cells into either CCR2+/+ or CCR2-/- nude mice. The expected results will provide further insight into the role of cytokines and chemokines and cellular migration in allergic asthma. This insight may allow us to devise novel therapeutics targeted at a specific molecule or at a specific cell population.

描述（申请人提供）：哮喘是一个重大的公共卫生问题，其发病机制复杂。 特别是，CC趋化因子在这种疾病中的作用知之甚少。 我们已经成功地开发了一种新的动物模型，其中抗原（Ag）脉冲的树突状细胞（DC）能够诱导气道高反应性（AHR），杯状细胞增生，特异性IgE反应，嗜酸性粒细胞增多，和显着的肺部炎症。 我们还表明，在CC趋化因子受体2（CCR 2-/-）缺陷的突变小鼠中，在吸入变应原后，在肺中观察到加速的Th 2应答。 其他人和我们已经表明，CCR 2缺陷改变单核细胞的迁移，并可能改变其他细胞，如Th淋巴细胞。 在该提议中，将研究MCP-1在诱导各种病理变化中的作用。 此外，将研究与哮喘发病机制相关的白细胞迁移改变。 提出了三个具体目标。 目标1：使用中和抗MCP-1抗体2 H5或CCR 2-/-/MCP-/-1双突变小鼠，证明升高的MCP- 1在Ag诱导的哮喘模型中观察到的肺中Th 2应答增强中起主要作用。 目的2：研究过敏原致敏的CCR 2-/-小鼠淋巴细胞、单核细胞和嗜酸性粒细胞向肺和气道迁移的动力学。 这将通过使用流式细胞术对总肺白细胞或肺组织的免疫组织化学染色在Ag激发开始后以增加的间隔研究小鼠来完成。 表征白细胞的抗体包括F4/80（巨噬细胞）、抗CD 4 mAb、抗CD 8 mAb、Thi（IFN高，IL-5低）、Th 2（IFN高，IL-5高）、抗MHC I类mAb（抗原呈递细胞）和抗DEC-205（DC）。目的2还将评估来自CCR 2-/-或CCR 2 +/+ DO 11.10小鼠的过继转移的Tg+ CD 4细胞的肺运输，以解决肺中Th亚群数量改变是否与运输或原位分化相关的问题。 目的3：确定单核细胞和淋巴细胞在过敏原攻击的CCR 2-/-小鼠中的Th 2应答增强中的相对作用。这将使用通过将CCR 2 +/+或CCR 2-/- Tg+ DO 11.10 CD 4细胞过继转移到CCR 2 +/+或CCR 2-/-裸鼠中而产生的嵌合小鼠来完成。 预期的结果将提供进一步了解细胞因子和趋化因子和细胞迁移在过敏性哮喘中的作用。 这一见解可能使我们能够设计针对特定分子或特定细胞群的新疗法。