THE V3 MASTOCYTOSIS MOUSE--MAST CELL DIFFERENTIATION, MATURATION, AND FUNCTION

已经知道一段时间了，驻留在不同 组织位置存储了不同类型和数量的颗粒 蛋白多糖和中性蛋白酶。尽管体外研究已经 哪些细胞因子可能调节肥大细胞颗粒 在BALB/c小鼠中的异质性，一直无法解决 小鼠肥大细胞的组织定向分化与成熟 紧张。它也不可能在实验中解决 体内颗粒异质性的功能意义。我们有 最近发现，当一只未成熟的v-abl转化的小鼠肥大 将V3-MC细胞过继转移到BALB/c小鼠体内， 肥大细胞增多症在肝、脾、肺和肠道迅速发展。 项目2的长期目标是使用V3肥大细胞增多症小鼠 研究肥大细胞的分化、成熟和功能；三 提出了具体的目标。初步数据显示，V3-MC 在这只患病小鼠的肝脏和肠道中发育 不同的颗粒表型。因此，蛋白水解酶特异性抗体和 将在特定目标1中使用基因特异性探针来研究 分化和成熟的组织导向调控 V3肥大细胞增多症小鼠的肥大细胞突起。因为纤维化是 通常与形态上的细胞数量增加有关 类似于成纤维细胞，采用转移系统将用于 特异性目的2研究肥大细胞介导的纤维化。V3-MC关联 肥大细胞增多症小鼠的肝和脾纤维化 与动物中的非纤维化组织进行比较，以确定 特定的肥大细胞表型调节宿主组织的反应。肥大细胞 含有比任何其他细胞都多得多的中性蛋白酶 尸体。然而，目前还不知道当这些蛋白水解酶 从体内的肥大细胞中排出。根据SDS的评估- PAGE/免疫印迹分析，高水平的小鼠肥大细胞蛋白酶7是 在V3肥大细胞增多症小鼠30分钟后出现在血液中 由免疫球蛋白E和抗原系统激活。因此，在具体目标3中， 培养的V3-3胞外不同颗粒成分的去向 MC和来自V3肥大细胞增多症小鼠的Fc-epsilon-RI 将对微扰进行调查。

It has been known for sometime that mast cells residing in different tissue locations store their granules varied types and amounts of proteoglycans and neutral proteases. Although in vitro studies have given in sight as to which cytokines probably regulate mast cell granule heterogeneity in the BALB/c mouse, it has not been possible to address tissue-directed mast cell differentiation and maturation in this mouse strain. It also has not been possible to experimentally address the functional significance of granule heterogeneity in vivo. We have recently discovered that when an immature v-abl-transformed mouse mast cell line (V3-MC) is adoptively transferred into a BALB/c mouse, systemic mastocytosis develops rapidly in the liver, spleen, lung, and intestine. The long term objective of project 2 is to use the V3 mastocytosis mouse to study mast cell differentiation, maturation, and function; three specific aims are proposed. Preliminary data indicate that the V3-MC that develop in the liver and intestine of this diseased mouse exhibit different granule phenotypes. Thus, protease specific antibodies and gene-specific probes will be used in specific aim 1 to investigate tissue-directed regulation of the differentiation and maturation processes of mast cells in V3 mastocytosis mice. Because fibrosis is usually associated with increased number of cells that morphologically resemble fibroblasts, the adoptive transfer system will be used in specific aim 2 to study mast-cell mediated fibrosis. V3-MC associated fibrosis in the liver and spleen of the mastocytosis mouse will be compared with non-fibrotic tissue in the animals to determine if a specific mast cell phenotype regulate host tissue responses. Mast cells contain substantially more neutral protease than any other cell in the body. However, it is not known what happens to these proteases when they are exocytosed from the mast cell in vivo. As assessed by SDS- PAGE/immunoblot analysis, high levels of mouse mast cell protease 7 are present in the blood 30 minutes after V3 mastocytosis mice are systemically activated with IgE and antigen. Thus in specific aim 3, the fate of the different granular constituents exocytosed from cultured V3- MC and from V3 mastocytosis mice with or without Fc-epsilon-RI perturbation will be investigated.