TTP PATHOGENESIS IN MODEL SYSTEMS IN VITRO AND IN VIVO

本研究提案是对NHLBI RFA HL-95-007（血栓性 血小板减少性紫癜（TTP）和HIV）。 长期目标是 进一步阐明TTP的发病机制， 实验模型，以研究可能涉及的因素， 血栓性微血管病（TMA）的发病机制，以确定 诱发因素、早期诊断和治疗见解。 有三个具体的目的：1）确定体外炎症和 最有可能参与凝血反应的细胞 TMA（内皮细胞、血小板和白细胞）的发病机制 例如志贺样毒素（SLT）和唾液酸酶的刺激剂。 使用 微血管和大血管内皮细胞的生长和比较 在静态和剪切应力条件下， 实验来自正常受试者、HIV和TTP患者的细胞和血浆 将用于阐明体外系统中的机制; 2） 使用动物（小鼠和小鼠）测定对SLT和唾液酸酶的体内应答。 已知对SLTs敏感的兔）。 特异性基因（P-选择素、E- 选择素、P-和E-选择素以及vWF）敲除小鼠将用于 确定我们观察的特异性; 3）前瞻性地 评价HIV中可能诱发TMA综合征的宿主因素 感染的病人。 这项研究将利用细胞和分子生物学技术， 研究毒素和唾液酸酶对各种感兴趣的细胞的影响。 的 这项研究的设计将使我们能够开始实验， 使用体外技术检查广泛的因素，然后 当我们找到相关的交互细胞时， 和分子。 从体外研究中获得的知识将 最终在一系列有针对性的体内实验中， 利用生理学方法研究整个动物的分子相互作用， 快递。 在项目完成后，我们将更多地了解 这些因素和细胞间的相互作用在发展中至关重要， TMA在HIV感染者和非感染者中。 这将提供 对诊断、治疗和预防发展的见解 养生法

This research proposal is a response to NHLBI RFA HL-95-007 (Thrombotic Thrombocytopenic Purpura (TTP) and HIV). The long term objectives are to further elucidate the pathogenesis of TTP using in vitro and in vivo experimental models to study factors that may be involved in the pathogenesis of thrombotic microangiopathy (TMA), to determine predisposing factors, early diagnosis and insights into treatment. There are three specific aims: 1) To determine in vitro inflammatory and coagulation responses of the cells most likely to be involved in the pathogenesis of TMA (endothelial cells, platelets and leukocytes) to known inciting agents such as shiga-like toxins (SLTs) and sialidase. The use of micro and macro vascular ECs and comparison of endothelial cells grown under static and shear stress conditions will add significantly to these experiments. Cells and plasma from normal subjects, HIV and TTP patients will be used to elucidate mechanisms in the in vitro system; 2) To determine in vivo responses to SLTs and sialidase using animals (mice and rabbits) known to be susceptible to SLTs. Specific gene (P-selectin, E- selectin, P- and E-selectin, and vWF) knockout mice will be used to determine the specificity of our observations; 3) To prospectively evaluate host factors potentially predisposing to TMA syndromes in HIV infected patients. The research will utilize cell and molecular biological techniques to study toxin and sialidase effects on the various cells of interest. The design of the study will allow us to initiate the experiments with an examination of a broad range of factors using in vitro techniques and then focus the experiments by degree as we find the relevant interactive cells and molecules. The knowledge gained from the in vitro studies will culminate in a series of targeted in vivo experiments looking at specific molecular interactions in whole animals utilizing physiological methods of SLT delivery. At the completion of the project we will know significantly more about the factors and cellular interactions which are critical in the development of TMA in both HIV infected and non-infected patients. This will provide insights for the development of diagnostic, therapeutic and preventative regimens.