Blockade of interleukin-6 signaling inhibits the classic pathway and promotes an alternative pathway of macrophage activation after spinal cord injury in mice

Journal of Neuroinflammation

Hisatoshi Baba⁴  William EB Johnson²  Masaya Nakamura¹  Shuji Watanabe⁴  Hideaki Nakajima⁴  Kenzo Uchida⁴  Alexander Rodriguez Guerrero³ 

[1] Department of Orthopaedic Surgery, School of Medicine, Keio University, Tokyo 160-8582, Japan;Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK;Servicio de Neurocirugia, Hospital Nacional Rosales, Universidad de El Salvador, San Salvador, El Salvador;Department of Orthopaedics and Rehabilitation Medicine, Faculty of Medical Sciences, University of Fukui, Matsuoka-Shimoaizuki 23, Eiheiji, Fukui 910-1193, Japan

关键词: T helper (Th) cytokine;    Inducible nitric oxide synthase (iNOS);    Arginase 1;    Alternatively activated macrophage;    Interleukin (IL)-6/IL6 receptor (R);    Spinal cord injury;   

Others  :  1220790
DOI  :  10.1186/1742-2094-9-40

 received in 2011-08-28, accepted in 2012-02-27,  发布年份 2012

【 摘 要 】

Background

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.

Methods

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.

Results

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.

Conclusion

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.

【 授权许可】

   
2012 Guerrero et al; licensee BioMed Central Ltd.

【 图 表 】

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