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CXCR2-positive neutrophils are essential for cuprizone-induced demyelination: relevance to multiple sclerosis

Nature Neuroscience volume 13, pages 319–326 (2010)Cite this article

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Abstract

Multiple sclerosis is an inflammatory demyelinating disorder of the CNS. Recent studies have suggested diverse mechanisms as underlying demyelination, including a subset of lesions induced by an interaction between metabolic insult to oligodendrocytes and inflammatory mediators. For mice of susceptible strains, cuprizone feeding results in oligodendrocyte cell loss and demyelination of the corpus callosum. Remyelination ensues and has been extensively studied. Cuprizone-induced demyelination remains incompletely characterized. We found that mice lacking the type 2 CXC chemokine receptor (CXCR2) were relatively resistant to cuprizone-induced demyelination and that circulating CXCR2-positive neutrophils were important for cuprizone-induced demyelination. Our findings support a two-hit process of cuprizone-induced demyelination, supporting the idea that multiple sclerosis pathogenesis features extensive oligodendrocyte cell loss. These data suggest that cuprizone-induced demyelination is useful for modeling certain aspects of multiple sclerosis pathogenesis.

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Figure 1: Cxcr2−/− mice are relatively resistant to cuprizone-induced demyelination.
Figure 2: Myelin protein mRNA expression is transiently reduced in Cxcr2−/− mice after cuprizone feeding and little apoptotic cell death occurs.
Figure 3: Differential response of cells of the oligodendrocyte lineage cells to cuprizone feeding in Cxcr2+/+ and Cxcr2−/− mice.
Figure 4: Cxcr2+/−→Cxcr2−/− and Cxcr2+/−→Cxcr2+/+ chimeric mice exhibited equal levels of cuprizone-induced demyelination.
Figure 5: Cxcr2−/−→Cxcr2+/+ mice are resistant to cuprizone-induced demyelination and there is a minimal reduction in the number of GST-π–positive cells or in the extent of the inflammatory reaction in corpus callosum of cuprizone-fed Cxcr2−/−→Cxcr2+/+ mice.
Figure 6: All Ly6G-positive neutrophils are CXCR2 positive and all CXCR2-positive cells are neutrophils.
Figure 7: Neutrophils infiltrate into the corpus callosum at the early stages of cuprizone-induced demyelination.
Figure 8: Neutrophils infiltrate in Cxcr2+/+ and Cxcr2−/− mice after 5 d of cuprizone feeding.

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Acknowledgements

We thank W. Stallcup for generously providing antibodies to PDGF receptor alpha and G. Kidd (Cleveland Clinic) for assistance with imaging. We thank G. Matsushima (University of North Carolina Chapel Hill) for helpful discussions regarding the cuprizone model. We thank X. Qu (Cleveland Clinic) for help with TUNEL staining. This research was supported by grants from the National Multiple Sclerosis Society (RG 3580 to R.M.R.), the US National Institutes of Health (NS32151 to R.M.R. and NS36674 to R.H.M.), the Myelin Repair Foundation (R.H.M.) and the Nancy Davis Center Without Walls (R.M.R.).

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Authors and Affiliations

  1. Department of Neuroscience, Neuroinflammation Research Center, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, USA

    LiPing Liu, Lindsey Darnall, Taofang Hu, Caitlin Drescher, Anne C Cotleur, Tao He, Karen Choi & Richard M Ransohoff

  2. Department of Neurosciences, Centers for Stem Cells and Regenerative Medicine, Translational Neuroscience, Case Western Reserve University, School of Medicine, Cleveland, Ohio, USA

    Abdelmadjid Belkadi, Dolly Padovani-Claudio & Robert H Miller

  3. Department of Molecular Biology & Biochemistry, University of California, Irvine, CA, USA

    Thomas E Lane

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Contributions

L.L. was responsible for day-to-day management of the project, designed and performed experiments, analyzed data, and wrote the initial draft of the manuscript. A.B. and R.H.M. designed and performed the experiments shown in Supplementary Figure 1. L.D., T. Hu and T. He were involved in generating and analyzing Cxcr2−/− mice on the B6 background. They performed experiments and were involved in data analysis and interpretation. C.D. participated in the development and implementation of electron microscopy quantitative methodology. A.C.C. interpreted flow cytometry experiments. D.P.-C. conducted initial LPC studies in Cxcr2−/− mice. K.C. quantified demyelination and assisted in the development of the technique. T.E.L. provided antibodies to CXCR2, participated in experimental design and provided input into the manuscript. R.H.M. participated in all aspects of experimental design and generation of the manuscript. R.M.R. conceived the project, designed the experiments and wrote the paper.

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Correspondence to Richard M Ransohoff.

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Liu, L., Belkadi, A., Darnall, L. et al. CXCR2-positive neutrophils are essential for cuprizone-induced demyelination: relevance to multiple sclerosis. Nat Neurosci 13, 319–326 (2010). https://doi.org/10.1038/nn.2491

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