ENVIRONMENTAL REGULATION OF NEUTROPHIL EXTRAVASATION

中性粒细胞从外周血管系统迁移到 牙周组织是宿主的保护性组织的重要组成部分， 对微生物菌斑积累的反应。神经元外渗 是通过局部释放炎性物质介导的 分子，内皮细胞和中性粒细胞表达粘附分子 细胞群，并刺激中性粒细胞的迁移， 释放趋化蛋白和脂质。自从改变了 中性粒细胞功能与牙周炎的发病机制有关， 疾病，整体而言。该提案的目标是制定一项 体外模型研究环境因素对 中性粒细胞外渗。牙周炎和 描述发生的任何有害后遗症。这些目标将 通过以下具体目标解决：（1）建立体外 使用双细胞膜的人血管壁的组织培养构建体 室系统，这将允许研究的粘附性和生理 中性粒细胞通过内皮、基底膜和型的迁移 I胶原蛋白对趋化蛋白和脂质的反应。(2)研究 中性粒细胞对粘附分子表达的调节， 在TNF-α存在下，内皮对趋化刺激的反应 a、IL-1、LPS、牙龈卟啉单胞菌或放线杆菌 伴放线菌;分析将使用血液的原位标记 血管壁构建体和单细胞悬浮液的FACS分析。 （三） 为了表征改变的粘附分子表达对 中性粒细胞的跨内皮迁移和 由于中性粒细胞的改变而发生的内皮细胞- 内皮细胞相互作用这些研究将提供重要的 关于特定炎症性疾病的环境影响的信息 中性粒细胞和内皮细胞相互作用的分子。 在 此外，这种体外构建体的开发将提供 为今后研究宿主和细菌的影响提供了极好的模型 炎症分子对牙周白细胞外渗的影响 疾病

The migration of neutrophils from the peripheral vasculature to the periodontal tissues is an essential component of the host's protective response to the accumulation of microbial plaque. Neutrophil extravasation to infected sites is mediated by the local release of inflammatory molecules, expression of adhesion molecules by endothelial and neutrophil cell populations, and the stimulated migration of neutrophils in response to the release of chemotactic proteins and lipids. Since altered neutrophil function has been implicated in the pathogenesis of periodontal disease, the overall. objectives of this proposal are to develop an in vitro model for studying the influence of environmental factors on neutrophil extravasation at sites of. periodontal inflammation and to characterize any deleterious sequelae that occur. These objectives will be addressed by the following specific aims: (1) To establish an in vitro tissue culture construct of the human blood vessel wall using a dual chamber system, that will allow the study of the adherence and physiologic migration of neutrophils through endothelium, basement membrane and type I collagen in response to chemotactic proteins and lipids. (2) To study the modulation of adhesiOn molecule expression by neutrophils and endothelium in response to chemotactic stimulation in the presence of TNF- a, IL-1, LPS, Porphyromonas gingivalis or Actinobacillus actinomycetemcomitans; analysis will use in situ labeling of the blood vessel wall construct and FACS analysis of single cell suspensions. (3) To characterize the effects of altered adhesion molecule expression on transendothelial migration of neutrophils and the pathologic changes in endothelial cells that occur as a result of the altered neutrophil- endothelial cell interaction. These studies will provide important information on the environmental effects of specific inflammatory molecules on the interaction of neutrophils and endothelial cells. In addition, the development of this in vitro construct will provide an excellent model for future studies of the effects of host and bacterial inflammatory,molecules on leukocyte extravasation in the periodontal diseases.