Chitosan IFNgamma-pDNA Nanosphere Therapy and Immunopathology of Allergic Asthma

描述(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，我们的目标是检测壳聚糖干扰素-伽马-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的有效性的细胞和分子机制。对过敏原的免疫反应从致病的T辅助-2型反应偏离为T辅助-1型，可能为改变病程提供了一种实用的方法。在草类过敏性哮喘小鼠模型中，给予干扰素-γ和IL-12DNA质粒组显著降低了呼吸道炎症和气道高反应性。此外，腺病毒介导的干扰素-γ基因转移有效地逆转了BALB/c小鼠模型中已建立的哮喘。虽然这些研究有助于了解干扰素-γ的机制，但在肺中的作用、急性炎症和对病毒的免疫原性仍然是病毒介导的基因转移应用于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，包括开发含有干扰素-γ的壳聚糖纳米球，PDNA(CIN)，滴鼻(I.N.)输送到肺部，作为治疗哮喘的一种策略。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表达的改变，导致抗原提呈改变，减少效应细胞向肺内的迁移，以及炎症细胞的凋亡，从而有效地预防或治疗哮喘，从而导致炎症和气道重塑的全面减少。本研究的具体目的如下：目的1.评价壳聚糖-干扰素-γ-PDNA纳米球(CIN)对BALB/c小鼠过敏性疾病的预防或治疗作用。我们计划从急性炎症的程度、哮喘的保护时间以及在慢性哮喘模型中的潜力来评估CIN在BALB/c小鼠过敏性哮喘预防/治疗中的作用。我们将分析不同的哮喘表型，如T细胞细胞因子分泌和抗体反应谱变化所揭示的变态反应的免疫偏离，支气管肺泡灌洗中的气道高反应性和嗜酸性粒细胞，以及肺病理。目的#2.阐明CIN诱导免疫调节的细胞/分子机制。我们计划研究T细胞的作用，CIN对肺内特异性T细胞反应的调节，包括Th2细胞的凋亡和肺内树突状细胞的数量和活性的调节。目的#3.阐明CIN保护作用的抗炎机制。我们计划检测介导CIN效应的基因，CIN是否影响小鼠肺内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大大有助于我们对哮喘的认识，CIN疗法可能在哮喘治疗方面提供重大突破。

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.