Seldin D C, Adelman S, Austen K F, Stevens R L, Hein A, Caulfield J P, Woodbury R G
Proc Natl Acad Sci U S A. 1985 Jun;82(11):3871-5. doi: 10.1073/pnas.82.11.3871.
Secretory granules of the rat basophilic leukemia (RBL-1) cell, a chemically generated tumor cell line maintained in tissue culture, were shown to stain with alcian blue but not with safranin counterstain and to have sparse, small, electron-dense granules. A Mr 25,000 protein was the major [3H]diisopropyl fluorophosphate-binding protein in extracts of RBL-1 cells. Double-immunodiffusion analysis of extracts revealed immunoreactivity for rat mast cell protease (RMCP)-II, a Mr 25,000 neutral protease present in the secretory granules of rat mucosal mast cells and cultured rat bone marrow-derived mast cells, but no immunoreactivity for RMCP-I, the predominant neutral protease of rat connective tissue mast cells. By radial immunodiffusion, there was 66.8 ng of RMCP-II per 10(6) cells. Whereas rat connective tissue mast cells stain with alcian blue and safranin and contain heparin proteoglycan, rat mucosal and rat bone marrow-derived mast cells stain with alcian blue only and contain a non-heparin proteoglycan and lesser amounts of histamine. Proliferation of rat mucosal mast cells in vivo and rat bone marrow-derived mast cells in vitro requires T-cell factors, whereas no comparable requirement has been observed for connective tissue mast cells. The transformed RBL-1 tumor cells, whose growth is independent of factors other than those present in standard tissue culture medium, has previously been shown to contain predominantly chondroitin sulfate di-B proteoglycans and low amounts of histamine. The similar histology and secretory granule biochemistry of the rat mucosal mast cells, rat culture-derived mast cell, and RBL-1 cell suggest that they comprise a single mast cell subclass distinct from the rat connective tissue mast cell.
大鼠嗜碱性白血病(RBL-1)细胞是一种在组织培养中维持的化学诱导肿瘤细胞系,其分泌颗粒经显示可被阿尔辛蓝染色,但不能被番红复染,且具有稀疏、小而电子致密的颗粒。一种分子量为25,000的蛋白质是RBL-1细胞提取物中主要的[3H]二异丙基氟磷酸结合蛋白。提取物的双向免疫扩散分析显示,其对大鼠肥大细胞蛋白酶(RMCP)-II具有免疫反应性,RMCP-II是一种分子量为25,000的中性蛋白酶,存在于大鼠黏膜肥大细胞和培养的大鼠骨髓来源肥大细胞的分泌颗粒中,但对大鼠结缔组织肥大细胞的主要中性蛋白酶RMCP-I没有免疫反应性。通过放射免疫扩散法,每10(6)个细胞中有66.8 ng的RMCP-II。大鼠结缔组织肥大细胞用阿尔辛蓝和番红染色,含有肝素蛋白聚糖,而大鼠黏膜和大鼠骨髓来源肥大细胞仅用阿尔辛蓝染色,含有非肝素蛋白聚糖和较少的组胺。大鼠黏膜肥大细胞在体内的增殖以及大鼠骨髓来源肥大细胞在体外的增殖需要T细胞因子,而未观察到结缔组织肥大细胞有类似需求。转化的RBL-1肿瘤细胞的生长不依赖于标准组织培养基中所含因子以外的其他因子,先前已显示其主要含有硫酸软骨素二-B蛋白聚糖和少量组胺。大鼠黏膜肥大细胞、大鼠培养来源肥大细胞和RBL-1细胞相似的组织学和分泌颗粒生物化学表明,它们构成了一个与大鼠结缔组织肥大细胞不同的单一肥大细胞亚类。