MECHANISMS OF IMMUNE COMPLEX INDUCED LUNG INJURY

三种大鼠急性肺损伤模型的建立(肺内 免疫球蛋白或免疫球蛋白A免疫复合体沉积，或全身激活后 ，我们将评估L-P-、E-选择素在这些过程中的作用 利用Ig-选择素嵌合体和阻断建立肺损伤模型 抗体。其次，使用新开发的方法来 ICAM-1、P-选择素和E-选择素的测定，我们将定量 在体内评估这些黏附分子在三种细胞中的上调 肺损伤模型。然后我们将确定在多大程度上 补体和细胞因子(TNFpha、IL-1)在体内是必需的 内皮细胞黏附分子表达上调。第三，我们将 延长初步研究表明IL-4具有强大的调节作用 和IL-10在肺内沉积后炎性损伤中的作用 Ig G或Ig A免疫复合体。我们将评估IL-4和IL-4的作用。 10对支气管肺泡灌洗液中肿瘤坏死因子α、白介素1含量的影响 和单核细胞趋化蛋白-1在免疫复合体诱导的肺损伤模型中的作用。这个 IL-4和IL-10对体内ICAM-1和E-2表达的影响 还将对选择素进行评估。同时，我们将在体外确定 IL-4和IL-10对血管内皮细胞转录上调的影响 内皮细胞ICAM-1、E-选择素、肿瘤坏死因子α、IL-1和单核细胞趋化蛋白-1与肺泡 巨噬细胞表达后三种细胞因子。第四，大鼠VCAM- 1将在杆状病毒系统和抗VCAM-1抗体中表达 将获得并在体内使用免疫复合体模型进行评估 VCAM-1的作用。第五，我们将继续进行体外研究， 大鼠肺泡I型上皮细胞的单层培养 β2整合素对大鼠肺泡黏附作用的要求 巨噬细胞以及这些黏附相互作用如何影响氧气的产生 巨噬细胞中的肿瘤坏死因子α。这些研究应该提供 关于粘附力作用的大量补充信息 肺损伤中的分子和细胞因子。最后，像动物一样 可用，使用肺内表达的转基因小鼠 C5a或IL-8/GRO同系物Kc，或使用带有C5a或Kc受体的小鼠 “击倒”，我们将确定这些手法对肺的影响 形态和补体依赖的肺部炎症反应(使用 在小鼠中，上述肺损伤模型。

Using three models of acute lung injury in rats (following intrapulmonary deposition of IgG or IgA immune complexes, or after systemic activation of complement), we will assess the roles of L-P-,E-selectins in these models of lung injury, employing Ig-selectin chimeras as well as blocking antibodies. Secondly, employing newly developed methods for the measurements of ICAM-1, and P- and E-selectins, we will quantitatively assess in vivo the upregulation of these adhesion molecules in the three models of lung injury. We will then determine the extent to which complement and cytokines (TNFalpha, IL-1) are required for the in vivo upregulation of the endothelial adhesion molecules. Thirdly, we will extend preliminary studies that suggest potent regulatory effect of IL-4 and IL-10 on inflammatory injury following intrapulmonary deposition of IgG or IgA immune complexes. We will assess the effects of IL-4 and IL- 10 on bronchoalveolar lavage (BAL) fluids for content of TNFalpha, IL-1 and MCP-1 in the models of immune complex-induced lung injury. The effects of IL-4 and IL-10 on in vivo upregulation of ICAM-1 and E- selectin will also be assessed. In parallel, we will determine in vitro the effects of IL-4 and IL-10 on transcriptional upregulation of endothelial ICAM-1, E-selectin, TNFalpha, IL-1 and MCP-1 and alveolar macrophage expression of the latter three cytokines. Fourthly, rat VCAM- 1 will be expressed in the baculovirus system and anti-VCAM-1 antibodies will be obtained and used in vivo in the immune complex models to assess the role of VCAM-1. Fifthly, we will pursue in vitro studies with monolayers of Type I rat alveolar epithelial cells in order to determine Beta 2 integrin requirements for adhesive interactions with rat alveolar macrophages and how these adhesive interactions affect production of O2 and TNFalpha in the macrophages. These studies should provide substantial, additional information regarding the role of adhesion molecules and cytokines in lung injury. Finally, as animals are available, employing transgenic mice with intrapulmonary expression of C5a or the IL-8/gro homologue, KC, or using mice with C5a or KC receptor "knockout", we will determine the effects of these manipulations on lung morphology and on complement dependent lung inflammatory reactions (using in mice the models of lung injury referred to above).