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Nerve growth factor-β induces mast-cell marker expression during in vitro culture of human umbilical cord blood cells

机译:神经生长因子-β在体外培养人脐带血细胞过程中诱导肥大细胞标志物表达

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摘要

Nerve growth factor-β (NGF) is known as a growth factor for human basophils and murine mast cells and has recently been shown to also up-regulate mast cell characteristics in human leukaemic mast cells. We have examined here the effect of NGF on the differentiation of normal human mast cells from cord blood progenitors during culture with stem cell factor (SCF), NGF alone or in combination, or fibroblast supernatants. All these supplements induced mast cell immunoreactivity against tryptase, c-Kit and FcεRIα, but none of the cells reacted against the basophil specific antibody 2D7 before or during culture. Intracellular tryptase activity increased as well, with maximal levels on combined culture with SCF and NGF. On reverse transcription–polymerase chain reaction (RT–PCR), cells lacked tryptase and chymase and expressed low levels of FcεRI and c-Kit mRNA prior to culture, with marked up-regulation of FcεRI and c-Kit, and with de novo expression of mast-cell specific α- and β-tryptase by week 3, and of chymase by week 5. Only the TrkA and not the p75 NGF receptor was detected at m-RNA and protein level, and only the TrkA NGF receptor was up-regulated during NGF-driven culture. These findings show therefore that, like SCF, NGF is another growth factor that can induce and regulate human mast-cell development and differentiation.
机译:神经生长因子-β(NGF)被称为人类嗜碱性粒细胞和鼠肥大细胞的生长因子,最近已被证明也可上调人类白血病肥大细胞的肥大细胞特性。我们在这里检查了NGF对干细胞因子(SCF),单独或联合使用NGF或成纤维细胞上清液培养过程中正常人肥大细胞从脐带血祖细胞分化的影响。所有这些补充剂均可诱导肥大细胞对类胰蛋白酶,c-Kit和FcεRIα的免疫反应,但在培养之前或培养过程中,没有细胞与嗜碱性粒细胞特异性抗体2D7反应。细胞内的类胰蛋白酶活性也增加了,与SCF和NGF联合培养的最大水平。在逆转录聚合酶链反应(RT-PCR)上,细胞在培养前缺乏类胰蛋白酶和糜酶,并表达低水平的FcεRI和c-Kit mRNA,具有明显的FcεRI和c-Kit上调,并且具有从头表达在第3周时检测出肥大细胞特异性α-和β-类胰蛋白酶,在第5周时检测出糜酶。在m-RNA和蛋白质水平上仅检测到TrkA而非p75 NGF受体,仅TrkA NGF受体在在NGF驱动的文化中受到调控。因此,这些发现表明,与SCF一样,NGF是另一个可以诱导和调节人类肥大细胞发育和分化的生长因子。

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