Blockade of interleukin-6 signaling inhibits the classic pathway and promotes an alternative pathway of macrophage activation after spinal cord injury in mice.
Blockade of interleukin-6 signaling inhibits the classic pathway and promotes an alternative pathway of macrophage activation after spinal cord injury in mice.
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DOI：
10.1186/1742-2094-9-40
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发表时间：
2012-02-27
影响因子：
9.3
通讯作者：
Baba H
中科院分区：
文献类型：
--
作者：
Guerrero AR;Uchida K;Nakajima H;Watanabe S;Nakamura M;Johnson WE;Baba H
最近在非神经元和神经元组织中的体内和体外研究表明，不同的巨噬细胞活化途径导致具有不同性质的细胞。白细胞介素（IL）-6触发经典活化的炎性巨噬细胞（M1表型），而替代活化的巨噬细胞（M2表型）是抗炎的。本研究的目的是阐明使用抗小鼠IL-6/IL-6受体（IL-6 R）单克隆抗体（MR 16 -1）暂时阻断IL-6/IL-6受体（IL-6 R）结合对小鼠脊髓损伤（SCI）后急性期巨噬细胞活化和炎症反应的影响。在小鼠胸部SCI后立即施用MR 16 -1抗体与同种型对照抗体或单独的盐水。通过Luxol固蓝（LFB）染色比较两组SC组织修复的髓鞘形成、神经元标记物生长相关蛋白（GAP）-43和神经丝重链200 kDa（NF-H）的免疫反应性以及运动功能。免疫印迹法检测Th 1细胞因子（IFN-γ和肿瘤坏死因子-α）和Th 2细胞因子（IL-4、IL-13）的表达。通过免疫组织学确定M1（诱导型一氧化氮合酶（iNOS）阳性，CD 16/32阳性）和M2（诱导型一氧化氮合酶1阳性，CD 206阳性）巨噬细胞的存在。使用流式细胞术，我们还定量了中性粒细胞、小胶质细胞和巨噬细胞中的IFN-γ和IL-4水平，以及M2巨噬细胞和小胶质细胞中的Mac-2（巨噬细胞抗原-2）和Mac-3水平。与对照组相比，MR 16 -1治疗组中LFB阳性备用髓鞘增加，并且这种增加与GAP-43或NF-H的阳性增强以及运动Basso小鼠量表评分改善相关。MR 16 -1处理的样品的免疫印迹分析鉴定了Th 1细胞因子的下调和Th 2细胞因子的上调。而iNOS阳性、CD 16/32阳性的M1巨噬细胞是未处理对照小鼠的损伤SC中的主要表型，MR 16 -1处理促进了CD 206阳性的M2巨噬细胞，这些细胞优先定位于损伤部位。MR 16 -1处理抑制了IFN-γ阳性中性粒细胞的数量，并增加了存在的小胶质细胞的数量及其对IL-4的阳性。在M2巨噬细胞中，MR 16 -1处理增加了Mac-2和Mac-3的阳性，表明吞噬行为增加。结果表明，SCI后暂时阻断IL-6信号传导可消除损伤性炎症活动，并通过促进替代性激活的M2巨噬细胞的形成来促进功能恢复。
Recent in vivo and in vitro studies in non-neuronal and neuronal tissues have shown that different pathways of macrophage activation result in cells with different properties. Interleukin (IL)-6 triggers the classically activated inflammatory macrophages (M1 phenotype), whereas the alternatively activated macrophages (M2 phenotype) are anti-inflammatory. The objective of this study was to clarify the effects of a temporal blockade of IL-6/IL-6 receptor (IL-6R) engagement, using an anti-mouse IL-6R monoclonal antibody (MR16-1), on macrophage activation and the inflammatory response in the acute phase after spinal cord injury (SCI) in mice. MR16-1 antibodies versus isotype control antibodies or saline alone were administered immediately after thoracic SCI in mice. SC tissue repair was compared between the two groups by Luxol fast blue (LFB) staining for myelination and immunoreactivity for the neuronal markers growth-associated protein (GAP)-43 and neurofilament heavy 200 kDa (NF-H) and for locomotor function. The expression of T helper (Th)1 cytokines (interferon (IFN)-γ and tumor necrosis factor-α) and Th2 cytokines (IL-4, IL-13) was determined by immunoblot analysis. The presence of M1 (inducible nitric oxide synthase (iNOS)-positive, CD16/32-positive) and M2 (arginase 1-positive, CD206-positive) macrophages was determined by immunohistology. Using flow cytometry, we also quantified IFN-γ and IL-4 levels in neutrophils, microglia, and macrophages, and Mac-2 (macrophage antigen-2) and Mac-3 in M2 macrophages and microglia. LFB-positive spared myelin was increased in the MR16-1-treated group compared with the controls, and this increase correlated with enhanced positivity for GAP-43 or NF-H, and improved locomotor Basso Mouse Scale scores. Immunoblot analysis of the MR16-1-treated samples identified downregulation of Th1 and upregulation of Th2 cytokines. Whereas iNOS-positive, CD16/32-positive M1 macrophages were the predominant phenotype in the injured SC of non-treated control mice, MR16-1 treatment promoted arginase 1-positive, CD206-positive M2 macrophages, with preferential localization of these cells at the injury site. MR16-1 treatment suppressed the number of IFN-γ-positive neutrophils, and increased the number of microglia present and their positivity for IL-4. Among the arginase 1-positive M2 macrophages, MR16-1 treatment increased positivity for Mac-2 and Mac-3, suggestive of increased phagocytic behavior. The results suggest that temporal blockade of IL-6 signaling after SCI abrogates damaging inflammatory activity and promotes functional recovery by promoting the formation of alternatively activated M2 macrophages.
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影响因子：
4.8
作者：
Bethea, JR;Dietrich, WD
通讯作者：
Dietrich, WD
DOI：
10.1523/jneurosci.3992-08.2009
发表时间：
2009-03-25
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Gensel JC;Nakamura S;Guan Z;van Rooijen N;Ankeny DP;Popovich PG
通讯作者：
Popovich PG
DOI：
10.1523/jneurosci.3257-09.2009
发表时间：
2009-10-28
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Kigerl KA;Gensel JC;Ankeny DP;Alexander JK;Donnelly DJ;Popovich PG
通讯作者：
Popovich PG
影响因子：
5.3
作者：
Gris, D;Marsh, DR;Weaver, LC
通讯作者：
Weaver, LC
影响因子：
16.2
作者：
DAVID, S;BOUCHARD, C;GIFTOCHRISTOS, N
通讯作者：
GIFTOCHRISTOS, N