Expression and regulation of interleukin-10 and interleukin-10 receptor in rat astroglial and microglial cells
Expression and regulation of interleukin-10 and interleukin-10 receptor in rat astroglial and microglial cells
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DOI：
10.1046/j.1460-9568.2002.02200.x
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发表时间：
2002-10-01
影响因子：
3.4
通讯作者：
Van Dam, AM
中科院分区：
文献类型：
--
作者：
Ledeboer, A;Brevé, JJP;Van Dam, AM
活化的神经胶质细胞对脑炎症反应至关重要。白细胞介素-10（IL-10）是脑中神经胶质细胞反应的重要调节剂。在本研究中，我们描述的表达IL-10和IL-10受体（IL-10 R1）在原代共培养的大鼠小胶质细胞和星形胶质细胞。利用定量RT-PCR和ELISA，我们表明，IL-10 mRNA的表达和随后的IL-10分泌的诱导脂多糖（LPS）的时间依赖性。然而，IL-10 R1在胶质细胞共培养物中组成型表达，如RT-PCR和免疫细胞化学所示。使用I-125-IL-10的放射性配体结合研究揭示，大鼠神经胶质细胞表达对人IL-10具有约600 pM表观亲和力的单一结合位点。在小胶质细胞或星形胶质细胞的富集培养物中的观察结果表明，两种细胞类型均表达IL-10 mRNA并能够分泌IL-10。两种细胞类型也表达IL-10 R1 mRNA和蛋白。然而，在神经胶质细胞共培养物中，免疫反应性IL-10 R1蛋白主要在星形胶质细胞中观察到，表明共培养物中小胶质细胞的IL-10 R1表达受到星形胶质细胞的抑制。此外，外源性IL-10在下调LPS诱导的IL-1 β和IL-10 mRNA方面非常有效，并且在较高剂量下，在未处理和LPS处理的培养物中下调IL-10 R1 mRNA，这表明IL-10在转录水平上自动调节其表达并抑制IL-1 β的表达。这些发现共同支持了这样的概念，即由活化的小胶质细胞和星形胶质细胞产生的IL-10通过旁分泌和自分泌相互作用通过高亲和力IL-10受体调节胶质细胞介导的炎症反应。
Activated glial cells crucially contribute to brain inflammatory responses. Interleukin-10 (IL-10) is an important modulator of glial cell responses in the brain. In the present study we describe the expression of IL-10 and the IL-10 receptor (IL-10R1) in primary cocultures of rat microglial and astroglial cells. Using quantitative RT-PCR and ELISA, we show that IL-10 mRNA expression and subsequent IL-10 secretion is time-dependently induced by lipopolysaccharide (LPS). IL-10R1, however, is constitutively expressed in glial cell cocultures, as shown by RT-PCR and immunocytochemistry. Radioligand binding studies using I-125-IL-10 reveal that rat glial cells express a single binding site with an apparent affinity of approximately 600 pM for human IL-10. Observations in enriched cultures of either microglial or astroglial cells indicate that both cell types express IL-10 mRNA and are capable of secreting IL-10. Both cell types also express IL-10R1 mRNA and protein. However, in glial cell cocultures immunoreactive IL-10R1 protein is predominantly observed in astrocytes, suggesting that microglial expression of IL-10R1 in cocultures is suppressed by astrocytes. In addition, exogenous IL-10 is highly potent in down-regulating LPS-induced IL-1beta and IL-10 mRNA, and, at a higher dose, IL-10R1 mRNA in untreated and LPS-treated cultures, suggesting that IL-10 autoregulates its expression and inhibits that of IL-1beta at the transcriptional level. Together the findings support the concept that IL-10, produced by activated microglial and astroglial cells, modulates glia-mediated inflammatory responses through high-affinity IL-10 receptors via paracrine and autocrine interactions.