Chitosan IFNgamma-pDNA Nanosphere Therapy and Immunopathology of Allergic Asthma

壳聚糖 IFNγ-pDNA 纳米球治疗过敏性哮喘的免疫病理学

• 批准号: 6803505
• 负责人: Shyam S Mohapatra
• 金额: $ 29万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-25 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6803505
• 关键词: T lymphocyte; allergens; antigen presentation; asthma; biotechnology; bronchial mucus; bronchomotion; chitin; diagnostic respiratory lavage; eosinophil; gene expression; gene therapy; genetically modified animals; goblet cells; immunopathology; immunotherapy; inhalation drug administration; interferon gamma; laboratory mouse; microcapsule; nanotechnology; nonhuman therapy evaluation; respiratory disease /disorder therapy; respiratory hypersensitivity; transfection /expression vector

DESCRIPTION (provided by applicant): With an intent to develop effective prophylaxis or treatment of allergic diseases including allergic asthma, our goal is to examine the safety and efficacy of chitosan IFN-gamma-pDNA nanosphere (CIN) Therapy and the cellular and molecular mechanisms underlying its effectiveness in a mouse model of asthma. Deviation of immune response to allergens from a pathogenic T helper-2 type response to T helper-1 type may provide a practical approach to modifying the course of disease. Administration of IFN-gamma, and IL-12 DNA plasmids significantly decreased airway inflammation and airway hyperresponsiveness in a mouse model of grass allergic asthma. In addition, Adenoviral-mediated IFN-gamma, gene transfer effectively reversed established asthma in BALB/c mouse model. While these studies aid in the mechanistic understanding of IFN-gamma, action in the lung, acute inflammation and immunogenicity to virus remains the major obstacle for the application of viral-mediated gene transfer for treating human asthma. We therefore developed a non-viral strategy that involves the development of chitosan nanospheres containing IFN-gamma, pDNA (CIN), intranasally (i.n.) delivered to the lung, as a strategy for asthma treatment. Our working hypothesis is that i.n. CIN therapy provides for effective prophylaxis or treatment of asthma by inducing changes in expression of cytokine and chemokine genes which result in altered antigen presentation, decreased migration of effector cells into the lung, and apoptosis of inflammatory cells, leading to a global decrease in inflammation and airway remodeling. The specific aims of this research program are as follows: Aim #1. Evaluate chitosan-IFN-gamma-pDNA nanospheres (CIN) as a prophylactic or a therapeutic modality for allergic disease in BALB/c mice. We plan to evaluate the role of CIN in prophylaxis/therapy of allergic asthma in BALB/c mice with respect to magnitude of acute inflammation, duration of protection from asthma and its potential in a chronic asthma model. We will analyze different asthma phenotypes, such as immune deviation of allergic response revealed by a change in T-cell cytokine secretion and antibody response profiles, the airway hyperreactivity and eosinophils in broncho-alveolar lavage, and lung pathology. Aim #2. Elucidate the cellular/molecular mechanism of CIN-induced immunomodulation. We plan to examine the role of T cells, CIN modulation of specific T cell response in the lung including apoptosis of Th2 cells and modulation of the number and activity of dendritic cells in the lung. Aim #3. Elucidate the anti-inflammatory mechanism of CIN-induced protection. We plan to examine the genes, which mediate the effects of CIN, whether CIN affects airway inflammation and airway remodeling in lungs of mice, and whether CIN induces apoptosis of mucus producing goblet cells. It is anticipated that the results of these studies will contribute significantly to our knowledge of asthma and CIN therapy may provide a major breakthrough in management of asthma.

描述（由申请人提供）：为了开发包括过敏性哮喘在内的过敏性疾病的有效预防或治疗方法，我们的目标是检查壳聚糖 IFN-γ-pDNA 纳米球 (CIN) 疗法的安全性和有效性，以及其在哮喘小鼠模型中有效性的细胞和分子机制。 对过敏原的免疫反应从致病性 T 辅助细胞 2 型反应转向 T 辅助细胞 1 型可能提供改变病程的实用方法。给予草过敏性哮喘小鼠模型中的 IFN-γ 和 IL-12 DNA 质粒可显着减少气道炎症和气道高反应性。 此外，腺病毒介导的 IFN-γ 基因转移有效逆转了 BALB/c 小鼠模型中已建立的哮喘。虽然这些研究有助于理解 IFN-γ 的机制，但其在肺部的作用、急性炎症和病毒的免疫原性仍然是应用病毒介导的基因转移治疗人类哮喘的主要障碍。因此，我们开发了一种非病毒策略，涉及开发含有 IFN-γ、pDNA (CIN) 的壳聚糖纳米球，通过鼻内 (i.n) 输送至肺部，作为哮喘治疗策略。我们的工作假设是 i.n. CIN疗法通过诱导细胞因子和趋化因子基因的表达变化来有效预防或治疗哮喘，从而导致抗原呈递改变、效应细胞向肺部迁移减少以及炎症细胞凋亡，从而导致炎症和气道重塑的全面减少。该研究计划的具体目标如下： 目标#1。评估壳聚糖-IFN-γ-pDNA 纳米球 (CIN) 作为 BALB/c 小鼠过敏性疾病的预防或治疗方式。我们计划评估 CIN 在 BALB/c 小鼠过敏性哮喘预防/治疗中的作用，包括急性炎症程度、哮喘保护持续时间及其在慢性哮喘模型中的潜力。我们将分析不同的哮喘表型，例如 T 细胞细胞因子分泌和抗体反应谱的变化所揭示的过敏反应的免疫偏差、支气管肺泡灌洗中的气道高反应性和嗜酸性粒细胞以及肺部病理学。目标#2。阐明 CIN 诱导的免疫调节的细胞/分子机制。我们计划研究 T 细胞的作用、CIN 对肺部特定 T 细胞反应的调节，包括 Th2 细胞的凋亡以及对肺部树突状细胞数量和活性的调节。目标#3。阐明 CIN 诱导保护的抗炎机制。我们计划检查介导CIN影响的基因，CIN是否影响小鼠肺部的气道炎症和气道重塑，以及CIN是否诱导产生粘液的杯状细胞的凋亡。预计这些研究的结果将极大地增进我们对哮喘的认识，CIN 治疗可能会为哮喘的治疗带来重大突破。