Matsuda H, Kannan Y, Ushio H, Kiso Y, Kanemoto T, Suzuki H, Kitamura Y
Department of Veterinary Surgery, College of Agriculture, University of Osaka Prefecture, Japan.
J Exp Med. 1991 Jul 1;174(1):7-14. doi: 10.1084/jem.174.1.7.
The effect of nerve growth factor (NGF) on proliferation/differentiation of mast cells was investigated in vitro. Although NGF alone neither supported colony formation of bone marrow-derived cultured mast cells (BMCMC) nor induced development of mast cell colonies from nonadherent bone marrow cells (NBMC), addition of NGF to the suboptimal dose of interleukin 3 (IL-3) significantly increased the numbers of mast cell colonies produced by BMCMC or NBMC in methylcellulose. When stimulated by IL-3 alone, cells in mast cell colonies were not stained by berberine sulfate, a fluorescent dye. In contrast, mast cells developing in methylcellulose cultures obtaining both IL-3 and NGF were stained by berberine sulfate. The fluorescence was abolished by the treatment of heparinase but not of chondroitinase ABC, suggesting that mast cells stimulated by IL-3 and NGF produced and stored heparin proteoglycan. The histamine content of BMCMC maintained by IL-3 was also increased by addition of NGF. Since BMCMC showed mucosal mast cell-like phenotype, NGF appeared to induce the phenotypic change to connective tissue-type mast cells (CTMC). In the culture containing BMCMC, 3T3 fibroblasts, and IL-3, the phenotypic change of BMCMC to CTMC was observed as well. Since NGF was detected in this coculture and since addition of anti-NGF monoclonal antibody suppressed the phenotypic change, NGF produced by fibroblasts appeared to induce the phenotypic change. Neither BMCMC alone nor IL-3 alone increased the concentration of NGF. Therefore, there is a possibility that BMCMC stimulated by IL-3 may induce the production and/or release of NGF by fibroblasts.
在体外研究了神经生长因子（NGF）对肥大细胞增殖/分化的影响。尽管单独的NGF既不支持骨髓来源的培养肥大细胞（BMCMC）的集落形成，也不诱导非贴壁骨髓细胞（NBMC）形成肥大细胞集落，但在亚最佳剂量的白细胞介素3（IL-3）中添加NGF可显著增加甲基纤维素中BMCMC或NBMC产生的肥大细胞集落数量。当仅由IL-3刺激时，肥大细胞集落中的细胞不会被硫酸小檗碱（一种荧光染料）染色。相反，在同时获得IL-3和NGF的甲基纤维素培养物中发育的肥大细胞被硫酸小檗碱染色。肝素酶处理可消除荧光，但软骨素酶ABC处理则不能，这表明由IL-3和NGF刺激的肥大细胞产生并储存了肝素蛋白聚糖。添加NGF也增加了由IL-3维持的BMCMC的组胺含量。由于BMCMC表现出黏膜肥大细胞样表型，NGF似乎诱导了向结缔组织型肥大细胞（CTMC）的表型转变。在含有BMCMC、3T3成纤维细胞和IL-3的培养物中，也观察到了BMCMC向CTMC的表型转变。由于在这种共培养物中检测到了NGF，并且添加抗NGF单克隆抗体可抑制表型转变，因此成纤维细胞产生的NGF似乎诱导了表型转变。单独的BMCMC或单独的IL-3均未增加NGF的浓度。因此，有可能由IL-3刺激的BMCMC可诱导成纤维细胞产生和/或释放NGF。
