MONONUCLEAR CELL FACTORS AFFECTING BASOPHILS/MAST CELLS

这项建议的目标是描述可溶因素的特征 由单核细胞释放，调节嗜碱性粒细胞和 肥大细胞。这些细胞在即刻超敏反应中起主要作用。 由IgE抗体引起的反应(哮喘、鼻炎、过敏反应)。 嗜碱性细胞也会在其他许多迟发反应中局部积聚： 接触性皮炎；肿瘤、微生物、移植的排斥反应； IgE依赖反应的慢性期。据推测， 单个核细胞释放细胞因子，引导嗜碱性粒细胞迁移 嗜碱性粒细胞和肥大细胞的分泌。我们已经确定并 描述了两种作用于嗜碱性粒细胞的人类细胞因子，一种嗜碱性细胞 趋化活性和组胺释放活性。 这些因子将通过培养白细胞来大量制备。 来自正常献血者和人类T细胞系的浓缩物。因素将 通过离子交换、凝胶过滤、亲和性和高性能进行纯化 液体色谱学。然后这些细胞因子将被用于生产 特异性抗体及其对嗜碱性粒细胞和肥大细胞的影响 细胞。多克隆抗体将通过免疫兔来制备 纯化的HRA，以提供一种用于检测单抗的方法 在小鼠体内制备。这些抗体将用于亲和力 HRA的层析、细胞因子的表征和测定 生物体液。 我们将比较未分离的单个核细胞产生的细胞因子。 与纯化的T细胞、T细胞亚群和 单核巨噬细胞。要进行比较的是物理化学 病毒的性质、抗原交叉反应性和功能反应 嗜碱性细胞。需要检验的一个假设是，相同的分子是否可以 调节嗜碱性粒细胞的迁移和分泌。我们推测，在 低浓度时，细胞因子诱导细胞运动，而在高浓度时， 浓度，胞吐作用。这将通过组合不同的 细胞因子与固相或液态抗体的浓度，然后 测定功能(趋化性和组胺释放)和 纯化的嗜碱性粒细胞的形态(电子显微镜)反应。 HRA的生物学意义将通过寻找这种细胞因子来研究。 在接触性皮炎患者的疱液中，以及在 哮喘患者的支气管肺泡灌洗液。自发的和 体外培养的哮喘患者细胞抗原诱导合成HRA 将遵循和水平与临床病程和 对免疫治疗的反应。

The objective of this proposal is to characterize the soluble factors released by mononuclear cells that regulate the function of basophils and mast cells. These cells have a major role in immediate hypersensitivity reactions (asthma, rhinitis, anaphylaxis) caused by IgE antibodies. Basophils also accumulate locally in many other late onset reactions: contact dermatitis; rejection of tumors, microbes, transplants; and the chronic stage of IgE dependent reactions. It is postulated that mononuclear cells release cytokines that direct the migration of basophils and the secretion of basophils and mast cells. We have identified and characterized two human cytokines acting on basophils, a basophil chemotactic activity and a histamine-releasing activity. These factors will be prepared in large quantities by culture of leukocyte concentrates from normal blood donors and human T-cell lines. Factors will be purified by ion exchange, gel filtration, affinity and high performance liquid chromatographies. The cytokines will then be used for production of specific antibodies and for determining their effects on basophils and mast cells. Polyclonal antibodies will be prepared by immunizing rabbits with purified HRA in order to provide an assay for monoclonal antibodies to be prepared in mice. These antibodies will be used for affinity chromatography, characterization of cytokines, and measurements of HRA in biological fluids. The cytokines produced by unfractionated mononuclear cells will be compared with the products of purified T-cells, T-cell subsets and monocyte-macrophages. Comparisons to be made are physicochemical properties, antigenic cross-reactivity and functional response of basophils. One hypothesis to be tested is whether the same molecules can regulate both migration and secretion of basophils. We speculate that at low concentrations, cytokines induce cell movement, and at high concentrations, exocytosis. This will be tested by combining different concentrations of cytokines with solid or fluid phase antibodies, and then determining the function (chemotaxis and release of histamine) and morphologic (electron microscopy) responses of purified basophils. The biological significance of HRA will be studied by seeking this cytokine in blister fluid from patients with contact dermatitis, and in bronchoalveolar lavage fluid from asthmatics. The spontaneous and antigen-induced synthesis of HRA by cells from asthmatics cultured in vitro will be followed and levels correlated with the clinical course and response to immunotherapy.