CHARACTERIZATION OF DIFFERENT TYPES OF HUMAN MAST CELLS

两种不同类型的人肥大细胞已被确定，并命名为T （类胰蛋白酶阳性，糜蛋白酶阴性）和TC（类胰蛋白酶阳性，糜蛋白酶 积极的）;拟议研究的目的是进一步表征 这些细胞类型。 将产生抗类胰蛋白酶的单克隆抗体， 糜蛋白酶和羧肽酶（肥大细胞中性蛋白酶）用于 更好地阐明这些酶的细胞分布， 免疫组织化学技术，并量化每种细胞水平 通过免疫测定。 分子RNA探针互补的mRNA，这些 将制备蛋白酶并用于检测该mRNA的表达 在成熟和未成熟的肥大细胞中。 合成的蛋白聚糖类型 每种细胞类型将通过分析35 S-SO 4标记的 蛋白聚糖由这些细胞的纯化制剂产生和释放。 单克隆抗体也将产生针对表面和 通过免疫接种这些细胞的细胞内成分， 来自成人的T和TC肥大细胞的分散/富集制剂， 新生组织 针对成人组织肥大细胞的抗体应 允许更好地定义这些肥大细胞的独特和共有的组分 类型 新生儿包皮中肥大细胞的抗体可以鉴定 未成熟肥大细胞的标记物，其将允许肥大细胞的前体 被检测和表征。 细胞是否是桅杆的证明 细胞前体将取决于在体外显示它可以发育 肥大细胞的分化特征，如组胺的存在 和类胰蛋白酶。 成熟型和未成熟型的超微结构特征 T和TC型肥大细胞以及肥大细胞的亚细胞定位 单克隆抗体识别的有趣抗原决定簇 将通过电子显微镜技术进行分析。 最后 T型和TC型肥大细胞的浓度和分布将是 确定无关的病理条件，包括类风湿性关节炎 关节炎、硬皮病、过敏性鼻炎、特应性皮炎、系统性 肥大细胞增多症/色素性荨麻疹、囊性纤维化、乳腺和肺实体 肿瘤和艾滋病。 因为T和TC型肥大细胞似乎分泌 共同的和不同的介质，沿着沿着不同的途径发展， 对某些非免疫促分泌素的反应不同， 药物制剂和不同的定位到不同的组织， 进一步的表征对于更好地理解 这些神秘的细胞对人类健康和疾病的影响。

Two different types of human mast cells have been identified and termed T (tryptase positive, chymase negative) and TC (tryptase positive, chymase positive); the purpose of the proposed research is to further characterize these cell types. Monoclonal antibodies will be produced against tryptase, chymase and carboxypeptidase (mast cell neutral proteases) to be used to better elucidate the cellular distribution of theses enzymes by immunohistochemical techniques and to quantify the cellular levels of each by immunoassay. Molecular RNA probes complementary to the mRNA of these proteases will be prepared and utilized to examine expression of this mRNA in mature and immature mast cells. The proteoglycan type synthesized by each cell type will be determined by analyzing the 35S-SO4 labeled proteoglycan produced and released by purified preparations of these cells. Monoclonal antibodies also will be produced against both surface and intracellular components of these cells by immunization with dispersed/enriched preparations of T and TC mast cells from adult and newborn tissues. Antibodies against mast cells form adult tissues should permit better definition of unique and shared components of these mast cell types. Antibodies against mast cells from newborn foreskin may identify markers of immature mast cells that will permit precursors of mast cells to be detected and characterized. Proof of whether or not a cell is a mast cell precursor will depend on showing in vitro that it can develop differentiated features of mast cells, such as the presence of histamine and tryptase. The ultrastructural features of mature and immature forms of T and TC types of mast cells as well as the subcellular localization of the interesting antigenic determinants recognized by the monoclonal antibodies will be analyzed by electron microscopic techniques. Finally, the concentration and distribution of T and TC types of mast cells will be determined indifferent pathologic conditions, including rheumatoid arthritis, scleroderma, allergic rhinitis, atopic dermatitis, systemic mastocytosis/urticaria pigmentosa, cystic fibrosis, breast and lung solid tumors and AIDS. Because T and TC types of mast cells appear to secrete both common and different mediators, develop along distinct pathways, respond differently to certain nonimmunologic secretagogues and pharmacologic agents and localize differently to different tissues their further characterization is crucial toward better understanding the role of these enigmatic cells to human health and disease.