HUMAN RECOMBINANT MIGRATION INHIBITORY FACTOR

淋巴细胞对抗原或促有丝分裂刺激的反应是分泌一种 一系列的冥想者，被称为淋巴因子， 免疫系统的细胞。 宏观经济在全球化中发挥着重要作用。 启动免疫应答和宿主防御，这些功能是 受多种淋巴因子控制 移动抑制因子 (MIF)，一种抑制巨噬细胞迁移的淋巴因子是 与细胞免疫相关性特别好的淋巴因子 应答 到目前为止，由于缺乏纯MIF， 它的物理和生物特性。 我们最近隔离了 对编码人MIF，MIF-1的cDNA进行测序（另外的独立克隆 已被分离，但尚未详细研究）。 它将允许 其基因和蛋白质结构的表征以及促进 阐明其生物活性。 本研究的目的是利用免疫荧光技术研究MIF-1基因的结构。 克隆MIF-cDNA作为探针筛选人基因组文库。 积极 将选择克隆用于分析内含子/外显子连接和5'和3'端 基因的区域和外显子的映射。 拟议的工作计划 采用凝胶过滤、Phenyl-Sepharose柱层析和Sepharose柱层析纯化重组MIF-1， 色谱法、等电聚焦和反相高压 液相色谱法和生产单克隆抗体，以重组 MIF-1使用已建立的方法。 本研究的目的是进一步研究重组MIF-1对肿瘤细胞增殖的影响。 培养的巨噬细胞和MIF-1与其他淋巴因子的相互作用 影响巨噬细胞 MIF在宿主防御和细胞免疫中的作用 免疫力将通过监测抗利什曼原虫活性来描述， 巨噬细胞单独或联合应用MIF的杀肿瘤活性 与其他淋巴因子结合。 这项研究还包括分析 编码不同MIF种类的额外cDNA克隆， 确定它们的DNA序列，以表征相应的MIF 蛋白质，以检查其生物活性，并将生产 特异性MIF种类对不同抗原刺激的反应， mRNA水平。

In response to antigenic or mitogenic stimulation, lymphocytes secrete an array of meditors, known as lymphokines which exert profound effects on cells of the immune system. Macrophages play important roles in the initiation of immune responses and in host defense and these functions are under the control of a variety of lymphokines. Migration inhibitory factor (MIF), a lymphokine that inhibits the migration of macrophages is one of the lymphokines that correlates especially well with cellular immune responses. Until now, the lack of pure MIF precluded the determination of its physical and biological properties. We have recently isolated and sequenced a cDNA encoding a human MIF, MIF-1 (additional independent clones have been isolated but not yet examined in detail). It will allow the characterization of its gene and protein structure as well as facilitate the elucidation of its biological activities. The proposed work aims to investigate the structure of MIF-1 gene using the cloned MIF-cDNA as a probe to screen a human genomic library. Positive clones will be selected for analysis of intron/exon junctions and 5' and 3' regions of the gene and for mapping of the exons. The proposed work plans to purify recombinant MIF-1 using gel filtration, Phenyl-Sepharose chromatography, isoelectric focusing, and reverse phase high pressure liquid chromatography and to produce monoclonal antibodies to recombinant MIF-1 using established methods. This study further aims to investigate the effects of recombinant MIF-1 on cultured macrophages and the interactions of MIF-1 with other lymphokines that affect macrophages. The role of MIF in host defense and in cellular immunity will be delineated by monitoring antileishmanial activity and tumoricidal activity of macrophages treated with MIF alone or in combination with other lymphokines., This study also includes the analysis of the additional cDNA clones encoding different MIF species in order to determine their DNA sequences, to characterize the corresponding MIF proteins, to examine their bioactivities, and to correlate the production of specific MIF species in response to different antigenic stimulation at the level of mRNA.