Reversible inhibitory effects of interferon-gamma and tumour necrosis factor-alpha on oligodendroglial lineage cell proliferation and differentiation in vitro
Reversible inhibitory effects of interferon-gamma and tumour necrosis factor-alpha on oligodendroglial lineage cell proliferation and differentiation in vitro
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DOI：
10.1111/j.1460-9568.1996.tb01278.x
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发表时间：
1996-06-01
影响因子：
3.4
通讯作者：
Levi, G
中科院分区：
文献类型：
--
作者：
Agresti, C;DUrso, D;Levi, G
我们研究了两种重要的炎症细胞因子，干扰素- γ (ifn - γ)和肿瘤坏死因子- α (tnf - α)对少突胶质细胞谱系细胞发育和存活的影响。从新生大鼠脑原代培养中获得纯化的少突胶质细胞和少突胶质细胞前体，在确定的无血清培养基中进行传代培养，并于第1天至第3天或第3天至第6天暴露于ifn - γ (1-100 U/ml), tnf - α (25-100 ng/ml)或两者（分别为100 U/ml和50 ng/ml）。虽然细胞存活在任何测试条件下都不受影响，但ifn - γ剂量依赖性抑制了[H-3]胸苷嘧啶或溴脱氧尿嘧啶的掺入（高达50%）和四唑盐3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四唑（MTT）的还原（高达33%）。tnf - α与ifn - γ协同作用，但本身无效。此外，ifn - γ完全拮抗碱性成纤维细胞生长因子和血小板源性生长因子对研究中少突胶质细胞系细胞群增殖的诱导作用。通过细胞形态学、早期和晚期分化标记物（分别为半乳糖脑苷和髓鞘碱性蛋白）的免疫染色以及神经酰胺半乳糖转移酶的活性，ifn - γ也阻断了少突胶质细胞前体的分化。ifn - γ的作用再次被tnf - α增强，而tnf - α在单独测试时无效。ifn - γ的抑制活性是快速可逆的：在去除细胞因子3天后，从第1天到第3天给药，可以证明细胞增殖和分化完全恢复。半定量逆转录-聚合酶链反应方法显示，细胞因子诱导的分化抗原表达阻滞伴随着相应mrna表达的扰动。特别是，在暴露于ifn - γ的培养物中，髓鞘碱性蛋白的信息（在治疗第3天至第6天的情况下，髓鞘相关糖蛋白的信息也是如此）减少，在ifn - γ和tnf - α处理的培养物中进一步降低，而tnf - α单独无效。上述观察结果可能有助于解释ifn - γ和tnf - α在炎性脱髓鞘疾病发病机制中的作用，其中这些物质水平的增加已被描述。特别是，在多发性硬化症的情况下，我们的结果可能与缺陷性髓鞘再生问题有关，并且与该疾病的频繁复发-缓解过程相一致。
We have investigated the effects of the two prominent inflammatory cytokines, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha), on oligodendroglial lineage cell development and survival. Purified oligodendrocytes and oligodendrocyte precursors obtained from neonatal rat brain primary cultures were subcultured in a defined, serum-free medium and exposed to IFN-gamma (1-100 U/ml), TNF-alpha (25-100 ng/ml) or both (100 U/ml and 50 ng/ml respectively) from day 1 to day 3 or from day 3 to day 6. While cell survival was not affected in any of the conditions tested, IFN-gamma dose-dependently inhibited [H-3]thymidine or bromodeoxyuridine incorporation (by up to 50%) and the reduction of the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; by up to 33%). TNF-alpha synergized with IFN-gamma but was ineffective by itself. Moreover, IFN-gamma totally antagonized the induction by basic fibroblast growth factor and platelet-derived growth factor of the proliferation of the oligodendroglial lineage cell population under study. IFN-gamma also blocked the differentiation of oligodendrocyte precursors, as evidenced by cell morphology, immunostaining for early and late differentiation markers (galactocerebroside and myelin basic protein respectively) and activity of ceramide galactosyl transferase. Again, the effect of IFN-gamma was potentiated by TNF-alpha, which was ineffective when tested alone. The inhibitory activity of IFN-gamma was rapidly reversible: 3 days after removal of the cytokine, administered from day 1 to day 3, complete recovery of cell proliferation and differentiation could be documented. The cytokine-induced arrest in the expression of differentiation antigens was accompanied by perturbations in the expression of the corresponding mRNAs, revealed by a semiquantitative reverse transcription-polymerase chain reaction method. In particular, the message for myelin basic protein (and, in the case of treatment from days 3 to 6, also that for myelin associated glycoprotein) was decreased in cultures exposed to IFN-gamma, and further depressed in cultures treated with IFN-gamma and TNF-alpha, while TNF-alpha alone was ineffective. The above observations may help explain the role of IFN-gamma and TNF-alpha in the pathogenesis of inflammatory demyelinating diseases, in which increases in the levels of these substances have been described. In particular, in the case of multiple sclerosis, our results may bear on the problem of defective remyelination and are consistent with the frequent relapsing-remitting course of the disease.