Diverse activation of microglia by chemokine (C-C motif) ligand 2 overexpression in brain.
Diverse activation of microglia by chemokine (C-C motif) ligand 2 overexpression in brain.
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DOI：
10.1186/1742-2094-10-86
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发表时间：
2013-07-17
影响因子：
9.3
通讯作者：
Gordon MN
中科院分区：
文献类型：
--
作者：
Selenica ML;Alvarez JA;Nash KR;Lee DC;Cao C;Lin X;Reid P;Mouton PR;Morgan D;Gordon MN
趋化因子(C-C基序)配体2(CCL2)是一种单核细胞趋化蛋白，在外周和中枢神经系统(CNS)炎症过程中介导巨噬细胞的募集和迁移。为了确定CCl_2在体内炎症中的作用，并阐明CCl_2对激活的脑小胶质细胞的极化作用，我们通过鸡β-肌动蛋白启动子驱动的重组腺相关病毒血清9型(rAAV-9)将CCl_2导入野生型小鼠的脑内。我们用组织学和化学方法测量小胶质细胞的激活，用组织学和流式细胞术测量单核细胞的募集。CCL2在中枢神经系统的过度表达诱导了脑内驻留的小胶质细胞的显著激活。CD45和主要组织相容性复合体II类免疫反应性在CCL2注射部位显著增加。小胶质细胞表型的组织学特征显示，小胶质细胞标志物如钙颗粒蛋白B和IL-1β以及与小胶质细胞交替激活相关的标志物如YM1和精氨酸酶1均升高。海马区蛋白表达谱显示，CCl_2可显著增加小胶质细胞中IL-6、GM-α和嗜酸性粒细胞趋化因子(CCL-11)的水平，而其他细胞因子包括肿瘤坏死因子-γ和干扰素-DNA的水平没有变化。此外，实时聚合酶链式反应分析证实，CCL2过表达后，与神经炎症相关的基因转录水平增加。最后，我们通过过继转移从无处不在表达绿色荧光蛋白的供体小鼠分离的骨髓来源细胞(BMDCs)来研究CCL2在体内的趋化特性。流式细胞术和组织学分析显示，BMDCs渗入脑实质，并与小胶质细胞标记物共标记。综上所述，我们的结果表明CCL2强烈激活了大脑中驻留的小胶质细胞。小胶质细胞的促炎和抗炎活性均显著，激活的细胞不偏向M1或M2表型。正如预期的那样，CCL2的过度表达积极地将循环单核细胞招募到中枢神经系统。因此，CCL2在小鼠脑内的表达可诱导小胶质细胞的激活，是一种有效的外周巨噬细胞募集方法。
The chemokine (C-C motif) ligand 2 (CCL2) is a monocyte chemoattractant protein that mediates macrophage recruitment and migration during peripheral and central nervous system (CNS) inflammation. To determine the impact of CCL2 in inflammation in vivo and to elucidate the CCL2-induced polarization of activated brain microglia, we delivered CCL2 into the brains of wild-type mice via recombinant adeno-associated virus serotype 9 (rAAV-9) driven by the chicken β-actin promoter. We measured microglial activation using histological and chemical measurement and recruitment of monocytes using histology and flow cytometry. The overexpression of CCL2 in the CNS induced significant activation of brain resident microglia. CD45 and major histocompatibility complex class II immunoreactivity significantly increased at the sites of CCL2 administration. Histological characterization of the microglial phenotype revealed the elevation of “classically activated” microglial markers, such as calgranulin B and IL-1β, as well as markers associated with “alternative activation” of microglia, including YM1 and arginase 1. The protein expression profile in the hippocampus demonstrated markedly increased levels of IL-6, GM-CSF and eotaxin (CCL-11) in response to CCL2, but no changes in the levels of other cytokines, including TNF-α and IFN-γ. Moreover, real-time PCR analysis confirmed increases in mRNA levels of gene transcripts associated with neuroinflammation following CCL2 overexpression. Finally, we investigated the chemotactic properties of CCL2 in vivo by performing adoptive transfer of bone marrow–derived cells (BMDCs) isolated from donor mice that ubiquitously expressed green fluorescent protein. Flow cytometry and histological analyses indicated that BMDCs extravasated into brain parenchyma and colabeled with microglial markers. Taken together, our results suggest that CCL2 strongly activates resident microglia in the brain. Both pro- and anti-inflammatory activation of microglia were prominent, with no bias toward the M1 or M2 phenotype in the activated cells. As expected, CCL2 overexpression actively recruited circulating monocytes into the CNS. Thus, CCL2 expression in mouse brain induces microglial activation and represents an efficient method for recruitment of peripheral macrophages.
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影响因子：
16.2
作者：
Bhaskar, Kiran;Konerth, Megan;Kokiko-Cochran, Olga N.;Cardona, Astrid;Ransohoff, Richard M.;Lamb, Bruce T.
通讯作者：
Lamb, Bruce T.
影响因子：
5.3
作者：
Cearley, Cassia N.;Wolfe, John H.
通讯作者：
Wolfe, John H.
影响因子：
3
作者：
Colton, Carol A.;Wilcock, Donna M.
通讯作者：
Wilcock, Donna M.
影响因子：
15.1
作者：
Glabinski, AR;Tani, M;Ransohoff, RM
通讯作者：
Ransohoff, RM
影响因子：
4.8
作者：
Elomaa, O;Sankala, M;Tryggvason, K
通讯作者：
Tryggvason, K