Thiol-sensitive mast cell lines derived from mouse bone marrow respond to a mast cell growth-enhancing activity different from both IL-3 and IL-4.
Thiol-sensitive mast cell lines derived from mouse bone marrow respond to a mast cell growth-enhancing activity different from both IL-3 and IL-4.
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源自小鼠骨髓的硫醇敏感肥大细胞系对不同于 IL-3 和 IL-4 的肥大细胞生长增强活性作出反应。
作者：
L. Hültner;J. Moeller;E. Schmitt;G. Jäger;G. Reisbach;J. Ring;P. Dörmer
用添加α-硫甘油的培养液和PWM刺激的脾细胞条件培养液作为IL-3的来源，从正常BALB/c或C3H骨髓中建立了一系列永久性的IL-3依赖细胞系。在琼脂中克隆的细胞系及其衍生物在表型和功能特性上类似于“粘膜型”肥大细胞。在这篇报道中，我们证明了这些肥大细胞株的体外生长不仅依赖于IL-3，并被IL-4协同促进，而且还受到α-硫代甘油的调节，后者可以被2-ME或半胱胺取代。我们发现，这些对硫醇敏感的肥大细胞系对脾细胞条件培养液中存在的肥大细胞生长促进活性(MEA)有反应，并与IL-3协同作用。部分纯化的MEA不能刺激IL-3依赖的32Dcl.23细胞、IL-2依赖的CTLL-2细胞或适应于纯化的IL-4的小鼠T细胞系F4/4K.6(L3T4+)的生长。此外，11B11杂交瘤产生的抗IL-4单抗中和小鼠IL-4不能取消MEA的生物活性。在我们的标准增殖试验中，PWM、CSF-1、GM-CSF、IL-1、IL-2、IL-5、IL-6、IL-7、干扰素-γ、转化生长因子-α、肿瘤坏死因子-α、神经生长因子或EPO不能替代MEA。
A series of permanent IL-3-dependent cell lines have been established from normal BALB/c or C3H bone marrow using alpha-thioglycerol-supplemented culture medium and PWM-stimulated spleen cell-conditioned medium as a source of IL-3. The cell lines and derivatives cloned in agar resembled "mucosal type" mast cells with respect to phenotypic and functional properties. In this report we demonstrate that in vitro growth of these mast cell lines was not only dependent on IL-3 and synergistically enhanced by IL-4, but in addition regulated by alpha-thioglycerol which could be replaced by 2-ME or cysteamine. We show that these thiol-sensitive mast cell lines respond to a mast cell growth enhancing activity (MEA) present in spleen cell-conditioned medium and acting in concert with IL-3. Partially purified MEA was not able to stimulate the growth of IL-3-dependent 32Dcl.23 cells, IL-2-dependent CTLL-2 cells or the mouse T cell line F4/4K.6 (L3T4+) adapted to grow in purified IL-4. Moreover, 11B11 hybridoma-derived anti-IL-4 mAb specifically neutralizing mouse Il-4 were unable to abolish the bioactivity of MEA. PWM, CSF-1, GM-CSF, IL-1, IL-2, IL-5, IL-6, IL-7, IFN-gamma, TGF-alpha, TNF-alpha, NGF, or EPO did not substitute for MEA in our standard proliferation assay.