Targeting macrophages by an aza-anthrapyrazole to ameliorate experimental autoimmune encephalomyelitis.
Targeting macrophages by an aza-anthrapyrazole to ameliorate experimental autoimmune encephalomyelitis.
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DOI：
10.1016/j.msard.2020.102190
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发表时间：
2020-05
影响因子：
4
通讯作者：
Boren Lin;Dylan Launder;Destiny Y. Bailey;Frank K Assifuah;Olivia A. Miller;H. Conti;Jianyang Du;Boyd M. Koffman
中科院分区：
文献类型：
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作者：
Boren Lin;Dylan Launder;Destiny Y. Bailey;Frank K Assifuah;Olivia A. Miller;H. Conti;Jianyang Du;Boyd M. Koffman
多发性硬化症（MS）是一种免疫介导的中枢神经系统神经退行性疾病，先天免疫细胞和适应性免疫细胞都参与其中。BBR3378是一种偶氮蒽吡唑，通过拮抗T细胞自身免疫反应来预防实验性自身免疫性脑脊髓炎（EAE），这是一种类似于MS的炎症。在这里，我们报道了BBR3378对巨噬细胞的调节作用。方法采用髓鞘少突胶质细胞糖蛋白肽免疫诱导10周龄雌性C57BL/6小鼠seae，然后腹腔注射BBR3378或假性治疗，采用0-5评分法评价临床症状。采用ELISA检测血清细胞因子IFNγ和TNFα水平，RT - qPCR检测分离单核细胞中巨噬细胞标志物，流式细胞术检测脑内巨噬细胞浸润情况。利用原代单核细胞和巨噬细胞系RAW 264.7提取的巨噬细胞，研究BBR3378对lps刺激的促炎细胞因子诱导的作用。表达NF-κB驱动的荧光素酶报告基因的RAW 264.7细胞分别用LPS（含或不含BBR3378）处理，荧光素酶测定评估对NF-κB活化的抑制作用。Western blot分析还研究了有或没有BBR3378存在的lps诱导的丝裂原活化蛋白激酶（MAPKs）的活化。结果bbr3378下调细胞因子诱导的EAE小鼠巨噬细胞分化和活化，对脑内巨噬细胞浸润和EAE临床症状有保护作用。用BBR3378处理巨噬细胞，通过阻断介导炎症反应的关键信号分子（如NF-κB和MAPKs）的激活，抵消lps诱导的细胞因子产生。结论BBR3378除可靶向T细胞外，还可靶向巨噬细胞减轻EAE相关炎症。
BackgroundMultiple sclerosis (MS) is an immune-mediated neurodegenerative disease in the central nerve system, in which both innate and adaptive immune cells are involved. BBR3378, an aza-anthrapyrazole prevents experimental autoimmune encephalomyelitis (EAE), an inflammatory condition similar to MS, by antagonizing T cell autoimmune responses. Here, we report BBR3378’s regulatory effect on macrophages.MethodsEAE was induced in ten-week-old female C57BL/6 mice by immunization with myelin oligodendrocyte glycoprotein peptides followed by BBR3378 or sham treatment administered intraperitoneally, and clinical signs were assessed using a 0–5 scoring system. These mice were subjected to serum ELISA for cytokine IFNγ and TNFα levels, RT qPCR analysis of macrophage markers in isolated monocytes, and flow cytometry analysis for macrophage infiltration in the brain. Macrophages derived from primary monocytes and macrophage cell line RAW 264.7 were used to investigate BBR3378’s effect on LPS-stimulated pro-inflammatory cytokine induction. RAW 264.7 cells expressing NF-κB-driven luciferase reporter were treated with LPS with or without BBR3378, and luciferase assays performed to assess the inhibition on NF-κB activation. LPS-induced activation of mitogen-activated protein kinases (MAPKs) with or without the presence of BBR3378 was also investigated by Western blot analysis.ResultsBBR3378 down-regulated cytokine-induced macrophage differentiation and activation in EAE mice, contributing to protection against macrophage infiltration in the brain and clinical symptoms from EAE. Treating macrophages with BBR3378 counteracted LPS-induced cytokine production via blocking activation of key signal molecules mediating inflammatory responses, such as NF-κB and MAPKs.ConclusionsThese data suggest that in addition to T cells, BBR3378 can also target macrophages to attenuate the inflammation associated with EAE.