LUNG INFLAMMATION

肺部炎症

基本信息

批准号：
2215286
负责人：
GERALD WEISSMANN
金额：
$ 32.19万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1979
资助国家：
美国
起止时间：
1979-06-01 至 1994-11-30
项目状态：
已结题

项目摘要

Neutrophils mediate tissue injury in two classical models of experimental pathology: the Arthus lesion(1903) and the Shwartzman phenomenon (1927). The aim of this proposal is to explain in the language of 1989 how vascular injury induced by immune complexes and complement (the Arthus lesion) differs from the induced by endotoxin, cytokines and complement (the Shwartzman phenomenon). We will study neutrophil/endothelial cell interactions in vitro to test novel hypotheses of "triggering" and activation" of neutrophils. We will determine the functional effects on human neutrophils and cytoplasts of "defensive" molecules release from activated vs. resting endothelial cells. A variety of endothelial cells (eg HUVEC) will be exposed to endotoxin or cytokines and the release measured of a) prostacyclin b) adenosine c) endothelium-derived relaxation factor (EDRF, or NO) and d) sphingosine (an endogenous inhibitor of O2 generation). The effects of these molecules will be studied on neutrophils or cytoplasts activated by chemoattractants or immune complexes with respect to a) heterotypic adherence (to HUVEC), b) cell-cell aggregation, c) disaggregation d) chemotaxis in several systems, and e) release of O-2 H2 O2 and lysosomal enzymes. We will also expose neutrophils activated by chemoattractants and immune complexes to PGI2 adenosine, NO and sphingosine. Stimulus/response coupling will be examined with attention to receptors for iC3b and immune complexes (FcRII,FcRIII), cytosolic pH and free calcium (pH, and Cai) 45Ca fluxes, membrane viscosity, membrane potential (delta psi), crosslinking of cytoskeletal proteins, microtubule assembly, phospholipid remodelling, and protein phosphorylation. We will compare the susceptibility of resting HUVEC with cells activated by endotoxin or cytokines (IL-1, TNF and IFN) to killing by neutrophils exposed to chemoattractants (C5a, LTB, FMLP). We will study whether immune complexes present a) in the bulk phase b) on the HUVEC monolayer, or c) in the subphase, provoke neutrophil-mediated cell injury despite release of "defensive" molecules from HUVEC. Finally, using marine sponge cells and human neutrophils, we will study whether 20.4 and other cis unsaturated fatty acids activate a novel GTP-binding protein the function of which is mimicked by phospatidic acid. These studies will employ novel liposomal methods for phospholipid presentation. Thus we will be able to define the roles played by cytokine activated endothelial cells (the Shwartzman model) and immune complexes (the Arthus model) in neutrophil-mediated blood vessel injury.

中性粒细胞介导了两个经典模型的实验模型病理学：Arthus病变（1903）和Shwartzman现象（1927）。该提议的目的是用1989年的语言解释免疫复合物和补体引起的血管损伤（Arthus 病变）与诱导的内毒素，细胞因子和补体不同（Shwartzman现象）。我们将研究中性粒细胞/内皮细胞体外相互作用以测试“触发”和激活“中性粒细胞。我们将确定对 “防御”分子的人类嗜中性粒细胞和细胞体从活化的与静息内皮细胞。多种内皮细胞（例如HUVEC）将暴露于内毒素或细胞因子和释放测量A）前列环蛋白b）腺苷c）内皮衍生松弛因子（EDRF，或NO）和D）鞘氨酸（内源性 O2代的抑制剂）。这些分子的影响将是在中性粒细胞或细胞体上进行了研究免疫复合物相对于a）异型依从性（对HUVEC），b）细胞细胞聚集，c）分类d）在几种系统，E）O-2 H2 O2和溶酶体酶的释放。我们也会暴露于化学吸引剂和免疫复合物激活的中性粒细胞 PGI2腺苷，NO和鞘氨醇。刺激/响应耦合将是对IC3B和免疫复合物的受体的关注进行了检查（FCRII，FCRIII），胞质pH和游离钙（pH和CAI）45CA通量，膜粘度，膜电位（Delta PSI），交联细胞骨架蛋白，微管组件，磷脂重塑，和蛋白质磷酸化。我们将比较用内毒素或细胞因子激活的细胞静止HUVEC（IL-1，TNF）和IFN）被暴露于趋化剂的中性粒细胞杀死（C5A，LTB， FMLP）。我们将研究免疫复合物是否存在a） B阶段B）在HUVEC单层上，或C）在子阶段中挑衅尽管释放了“防御性”分子，但中性粒细胞介导的细胞损伤来自Huvec。最后，使用海洋海绵细胞和人类嗜中性粒细胞，我们将研究20.4和其他顺式不饱和脂肪酸是否激活A 新型GTP结合蛋白的功能被模仿凤凰酸。这些研究将采用新颖的脂质体方法磷脂表现。因此，我们将能够定义角色由细胞因子激活的内皮细胞（Shwartzman模型）和中性粒细胞介导的血管中的免疫复合物（Arthus模型）受伤。