CHARACTERIZATION OF DIFFERENT TYPES OF HUMAN MAST CELLS

已鉴定出两种不同类型的人类肥大细胞，命名为T (类胰酶阳性，糜酶阴性)和TC(类胰酶阳性，糜酶阴性) 积极)；拟议研究的目的是进一步确定 这些细胞类型。将产生针对类胰蛋白酶的单抗， 乳糜酶和羧基肽酶(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个细胞的水平 通过免疫分析。与这些基因的mRNA互补的分子RNA探针 将制备蛋白水解酶并用于检测该基因的表达。 在成熟和未成熟的肥大细胞中。合成的蛋白多糖型 每种细胞类型将通过分析标记的35S-SO4来确定 蛋白多糖由这些细胞的纯化制剂产生和释放的蛋白多糖 还将产生针对表面和表面的单抗 通过免疫这些细胞的胞内成分 成人和成人T和TC肥大细胞的分散/浓缩制剂 新生儿的纸巾。来自成人组织的针对肥大细胞的抗体应该 允许更好地定义这些主单元的唯一和共享组件 类型。针对新生儿包皮肥大细胞的抗体可以识别 未成熟肥大细胞的标志物，将允许肥大细胞的前体细胞 被检测和表征。单元格是否为桅杆的证明 细胞前体将依赖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类型的肥大细胞以及肥大细胞的亚细胞定位 由单抗识别的有趣的抗原决定簇 将通过电子显微镜技术进行分析。最后， T和TC型肥大细胞的浓度和分布 在不同的病理条件下确定的，包括类风湿 关节炎，硬皮病，过敏性鼻炎，特应性皮炎，全身性 肥大细胞增多症/色素性荨麻疹，囊性纤维化，乳房和肺实性 肿瘤和艾滋病。因为T和TC型肥大细胞似乎能分泌 既有共同的也有不同的媒介，沿着不同的途径发展， 对某些非免疫性促分泌剂的反应不同 不同组织的药理作用和定位不同 进一步的表征对于更好地理解 这些神秘的细胞对人类的健康和疾病都有影响。

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.