NOVEL PROINFLAMMATORY CYTOKINE AND ENDOTHELIUM

内皮细胞(EC)凝血剂、屏障、粘附剂和 增殖功能是炎症的核心，而宿主的反应 到肿瘤。我们已经鉴定、克隆和表征了一部小说 多肽，命名为内皮-单核细胞激活多肽II (EMAPII)，它调节这些EC功能，并具有生理性和 血管生成。活化的小鼠单核细胞合成EMAPII 吞噬细胞(MPS)和构成小鼠Meth A肉瘤(Meth A) 细胞，并以约18-20kDa的单链分子形式分泌。 EMAPII还激活两种MPS，刺激细胞迁移和组织 因子合成和中性粒细胞(PMN)，促进 趋化性和超氧化物的产生。活体内，皮下 注射EMAPII引起炎症；但Meth A肿瘤细胞 组成型过表达的EMAPII基因表达增加 成长。我们假设EMAPII的背景和动力学 产品决定了它的生物学特性，无论是作为造影剂 在炎症环境中，或在肿瘤中作为新血管生成的诱导剂。 具体目的是了解EMAPII引发 炎症，并影响肿瘤宿主的反应。使用cdna探针和 抗EMAPII(鼠和人)的抗体，我们将确定 在一系列肿瘤中合成和聚集，炎性和 血管损伤。近邻成熟的EMAPII区的功能活性 Hn2末端，在合成肽中，刺激PMN和MP 迁移，特异性地结合到细胞，并与大约73 KDA细胞表面多肽将使用额外的多肽进行检测 和定点突变。这些试剂还将用于 鉴定并分离EMAPII细胞表面结合位点。基于 EMAPII和EMAPII的NH2-末端区序列同源性较高 血管性血友病抗原II(VWAgII)是一种存在于血小板中的多肽。 颗粒并由受刺激的内皮细胞释放)，我们已经表明vWAgII具有 属性类似于EMAPII。因为分泌的血小板的作用 蛋白质在组织修复中的作用，我们将评估vWAgII对内皮细胞的影响， MPS和PMN，并确定其在炎症和创伤中的作用 在动物模型中修复。由于vWAgII水平升高，因此 精氨酸加压素类似物DDAVP诱导的MP组织 因素，我们将知道vWAgII是否能够启动 体内促凝血机制的激活与血栓形成 使DDAVP治疗复杂化。这些研究提供了新的联系方式 凝血与炎症，并定义一种新的配体-受体 涉及EMAPII的互动，与宿主在 炎症、肿瘤和血管病变，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.