Quentmeier H, Kolsdorf K, Zaborski M, Drexler H G
DSM-German Collection of Microorganisms and Cell Cultures, Department of Human and Animal Cell Cultures, Braunschweig.
Leukemia. 1995 Feb;9(2):336-40.
Short-term stimulation of peripheral blood monocytes (PBMo) and cells of the monocytic cell line MONO-MAC-6 with lipopoly-saccharide (LPS) induces high tumor necrosis factor (TNF)alpha mRNA levels. In contrast to the results obtained with primary cells, this effect could not be inhibited by preincubating the cell line with recombinant human interleukin-4 (rh IL-4). This deficiency in response to the cytokine was not caused by a general unresponsiveness of MONO-MAC-6 cells to IL-4. Thus, the expression of the monocyte-associated differentiation markers CD14 and monocyte-specific esterase (MSE), upregulated by long-term stimulation with LPS, could be decreased by IL-4. Long-term LPS treatment apparently induced IL-4 responsiveness of the cell line. While IL-4R alpha mRNA was upregulated about 3-fold, this positive effect was not apparent at the cell surface protein level. In contrast to the constitutive alpha chain expression, the IL-4R gamma chain expression could not be detected with a specific mAb nor by Northern blot analysis. However, reverse transcriptase polymerase chain reaction (RT-PCR) demonstrated the presence of low-level IL-4R gamma chain mRNA in the cell line. We suggest that the low reactivity of the cells to IL-4 might be correlated with the low expression of the gamma chain.
用脂多糖(LPS)对人外周血单核细胞(PBMo)和单核细胞系MONO-MAC-6细胞进行短期刺激,可诱导肿瘤坏死因子(TNF)α mRNA水平升高。与原代细胞的结果不同,用重组人白细胞介素-4(rh IL-4)预孵育该细胞系并不能抑制这种效应。这种对细胞因子反应的缺陷并非由MONO-MAC-6细胞对IL-4普遍无反应所致。因此,长期用LPS刺激上调的单核细胞相关分化标志物CD14和单核细胞特异性酯酶(MSE)的表达,可被IL-4降低。长期LPS处理显然诱导了该细胞系对IL-4的反应性。虽然IL-4Rα mRNA上调了约3倍,但这种正向效应在细胞表面蛋白水平并不明显。与组成型α链表达不同,用特异性单克隆抗体或Northern印迹分析均未检测到IL-4Rγ链的表达。然而,逆转录聚合酶链反应(RT-PCR)证明该细胞系中存在低水平的IL-4Rγ链mRNA。我们认为细胞对IL-4的低反应性可能与γ链的低表达有关。