Detailed analysis of inflammatory and neuromodulatory cytokine secretion from human NT2 astrocytes using multiplex bead array
Detailed analysis of inflammatory and neuromodulatory cytokine secretion from human NT2 astrocytes using multiplex bead array
复制标题
DOI：
10.1016/j.neuint.2011.09.002
复制
发表时间：
2012-05-01
影响因子：
4.2
通讯作者：
Graham, E. Scott
中科院分区：
文献类型：
--
作者：
Burkert, Kristina;Moodley, Kiebashne;Graham, E. Scott
星形胶质细胞是大脑中一种非常重要的细胞类型，在神经免疫学和神经传递中发挥着重要的作用。我们对分泌的细胞因子(任何来源的星形胶质细胞)进行了迄今为止最全面的分析，以确定来自人类Ntera2(NT2)细胞系的星形胶质细胞是否是人类原代星形胶质细胞的良好模型。我们比较了NT2星形胶质细胞分泌的细胞因子与富含星形胶质细胞的人脑培养物产生的细胞因子，以及与脑损伤有关的或已知在人脑中表达的额外细胞因子。用多道微珠阵列(MBA)检测星形胶质细胞条件培养液中细胞因子的浓度，同时检测18种细胞因子。在非炎症条件下，静息的NT2星形胶质细胞产生低水平的MIP1α、IL-6和GM-CSF(接近1-30pg/ml)，高水平的MCP-1、IP-10和IL-8(1-11 ng/ml)。除IL-1β、TNF-α和IL-13外，所有这些都通过促炎激活(肿瘤坏死因子α或IL-1β刺激)而增加。而星形胶质细胞条件培养液中未检测到IL-2、IL-4、IL-5、IL-7、IL-10、IL-12、LTα和干扰素-γ。NT2星形胶质细胞对IL-2和腺苷环化酶激动剂Forsklin无反应。有趣的是，干扰素刺激只选择性地增加IP-10的分泌。当星形胶质细胞在IL-1β或肿瘤坏死因子α刺激下产生几种在ng/ml范围内的趋化因子时，我们接下来评估这些细胞的趋化特性。经肿瘤坏死因子α刺激的星形胶质细胞条件培养液可显著吸引人血中的白细胞。这项研究提供了迄今为止对人类星形胶质细胞产生细胞因子的最全面的分析，并表明NT2星形胶质细胞对包括TNFα和IL-1β在内的促炎介质具有高度的反应性，产生能够吸引人血液中的白细胞的细胞因子和趋化因子。我们的结论是，在没有成人原代星形胶质细胞的情况下，NT2-星形胶质细胞可能为研究人类星形胶质细胞的免疫学行为提供了一个有价值的替代方案。(C)爱思唯尔有限公司出版的2011年。
Astrocytes are a very important cell type in the brain fulfilling roles in both neuroimmunology and neurotransmission. We have conducted the most comprehensive analysis of secreted cytokines conducted to date (astrocytes of any source) to determine whether astrocytes derived from the human Ntera2 (NT2) cell-line are a good model of human primary astrocytes. We have compared the secretion of cytokines from NT2 astrocytes with those produced in astrocyte enriched human brain cultures and additional cytokines implicated in brain injury or known to be expressed in the human brain. The concentration of cytokines was measured in astrocyte conditioned media using multiplex bead array (MBA), where 18 cytokines were measured simultaneously. Resting NT2 astrocytes produced low levels (similar to 1-30 pg/ml) of MIP1 alpha, IL-6 and GM-CSF and higher levels of MCP-1, IP-10 and IL-8 (1-11 ng/ml) under noninflammatory conditions. All of these in addition to IL-1 beta, TNF alpha, and IL-13, were increased by pro-inflammatory activation (TNF alpha or IL-1 beta stimulation). In contrast, IL-2, IL-4, IL-5, IL-7, IL-10, IL-12, LT alpha, and IFN gamma were not detected in astrocyte conditioned media under any of the culture conditions tested. NT2 astrocytes were unresponsive to IL-2 and the adenyl cyclase agonist, forskolin. Interestingly, IFN gamma stimulation selectively increased IP-10 secretion only. As astrocytes stimulated with IL-1 beta or TNF alpha produced several chemokines in the ng/ml range, we next assessed the chemoattractant properties of these cells. Conditioned media from TNF alpha-stimulated astrocytes significantly chemoattracted leukocytes from human blood. This study provides the most comprehensive analysis of cytokine production by human astrocytes thus far, and shows that NT2 astrocytes are highly responsive to pro-inflammatory mediators including TNF alpha and IL-1 beta, producing cytokines and chemokines capable of attracting leukocytes from human blood. We conclude that in the absence of adult human primary astrocytes that NT2-astrocytes may provide a valuable alternative to study the immunological behaviour of human astrocytes. (C) 2011 Published by Elsevier Ltd.