NOVEL PROINFLAMMATORY CYTOKINE AND ENDOTHELIUM

调节内皮细胞（EC）的凝血、屏障、粘附和 增殖功能是炎症和宿主反应的核心 肿瘤。 我们鉴定、克隆并鉴定了一种新的 一种多肽，命名为内皮-单核细胞激活多肽II （EMAPII），其调节这些EC功能，并且是致炎的， 血管生成 EMAPII由活化的鼠单核细胞合成， 巨噬细胞（MP）和组成型小鼠甲硫氨酸肉瘤（甲硫氨酸） 细胞，并分泌为约18-20 kDA的单链分子。 EMAPII还激活MP，刺激细胞迁移和组织 因子合成和多形核白细胞（PMNs），促进 趋化性和超氧化物生成。 体内，皮下 给予EMAPII缓解炎症;但Meth A肿瘤细胞 转染以组成型过表达EMAPII， 增长 我们假设EMAPII的背景和动力学 生产决定其生物学特性，无论是作为致炎剂 在炎症环境中，或作为肿瘤中新血管生成的诱导剂。 具体的目的是了解EMAPII的机制， 炎症，并影响肿瘤宿主反应。 使用cDNA探针和 抗体EMAPII（鼠和人），我们将确定的网站， 在一系列肿瘤、炎症和 血管病变 成熟EMAPII近端区域的功能活性 HN 2-末端，在合成肽中，刺激PMN和MP 迁移，特异性结合细胞，并交联到约73 将使用其他肽检查kDa细胞表面多肽 和定点诱变。 这些试剂也将用于 表征并分离EMAPII细胞表面结合位点。 基于 EMAPII的NH 2-末端区域和 血管性血友病抗原II（vWAg II，血小板α- 颗粒和刺激EC释放），我们已经表明vWAg II具有 类似EMAPII的特性。 由于分泌血小板的作用， 组织修复中的蛋白质，我们将评估vWAgII对EC的影响， MP和PMNs，并确定其对炎症和创伤的贡献 修复动物模型。 由于vWAg II水平升高， 精氨酸加压素类似物DDAVP诱导的MP组织 因素，我们将查明vWAgII是否能够启动 体内促凝血机制的激活和血栓形成 使DDAVP治疗复杂化。 这些研究提供了新的方法， 凝血与炎症，并定义一个新的配体受体 相互作用，涉及EMAPII，与宿主反应相关， 炎症、瘤形成和血管病，如动脉粥样硬化。

Regulation of endothelial cell (EC) coagulant, barrier, adhesive and proliferative functions is central to inflammation, and the host response to tumors. We have identified, cloned and characterized a novel polypeptide, designated Endothelial-Monocyte Activating Polypeptide II (EMAPII), which modulates these EC functions, and is phlogogenic and angiogenic. EMAPII is synthesized by activated murine mononuclear phagocytes (MPs) and constitutively by murine Meth A sarcoma (Meth A) cells, and is secreted as an about 18-20 kDA single chain molecule. EMAPII also activates both MPs, stimulating cell migration and tissue factor synthesis, and polymorphonuclear leukocytes (PMNs), promoting chemotaxis and superoxide generation. In vivo, subcutaneously administered EMAPII elicits inflammation; but Meth A tumor cells transfected to constitutively overexpress EMAPII exhibit increased growth. We hypothesize that the context and kinetics of EMAPII production determine its biologic properties, either as phlogogenic agent in inflammatory settings, or as inducer of neoangiogenesis in tumors. The specific aims are to understand mechanisms by which EMAPII elicits inflammation, and influences tumor-host reactions. Using cDNA probes and antibodies to EMAPII (murine and human), we will identify sites of synthesis and accumulation in a spectrum of tumors, inflammatory and vascular lesions. Functional activity of a region of mature EMAPII near the HN2-terminus, which, in synthetic peptides, stimulates PMN and MP migration, binds specifically to cells, and cross-links to an about 73 kDa cell surface polypeptide will be examined using additional peptides and site-directed mutagenesis. These reagents will also be used to characterize and isolate the EMAPII cell surface binding site. Based on strong sequence homology between the NH2-terminal region of EMAPII and von Willebrand antigen II (vWAgII, a polypeptide in platelet alpha- granules and released by stimulated ECs), we have shown that vWAgII has properties resembling EMAPII. Because of the role of secreted platelet proteins in tissue repair, we will assess the effects of vWAgII on ECs, MPs, and PMNs, and determine its contribution to inflammation and wound repair in animal models. And since elevated levels of vWAgII, consequent on infusion of the arginine vasopressin analog, DDAVP, induced MP tissue factor, we will find out whether vWAgII is capable of initiating activation of procoagulant mechanisms in vivo, and the thrombosis complicating DDAVP therapy. These studies offer new means of linking coagulation with inflammation, and define a new ligand-receptor interaction, involving EMAPII, relevant to host responses in inflammation, neoplasia and, vasculopathies, such as atherosclerosis.