Abstract
Inflammation plays a critical role in the pathogenesis of ischemic stroke. This process depends, in part, upon proinflammatory factors released by activated resident central nervous system (CNS) microglia (MG). Previous studies demonstrated that transfer of IL-10+ B-cells reduced infarct volumes in male C57BL/6 J recipient mice when given 24 h prior to or therapeutically at 4 or 24 h after experimental stroke induced by 60 min middle cerebral artery occlusion (MCAO). The present study assesses possible sex differences in immunoregulation by IL-10+ B-cells on primary male vs. female MG cultured from naïve and ischemic stroke-induced mice. Thus, MG cultures were treated with recombinant (r)IL-10, rIL-4 or IL-10+ B-cells after lipopolysaccharide (LPS) activation and evaluated by flow cytometry for production of proinflammatory and anti-inflammatory factors. We found that IL-10+ B-cells significantly reduced MG production of TNF-α, IL-1β and CCL3 post-MCAO and increased their expression of the anti-inflammatory M2 marker, CD206, by cell-cell interactions. Moreover, MG from female vs. male mice had higher expression of IL-4 and IL-10 receptors and increased production of IL-4, especially after treatment with IL-10+ B-cells. These findings indicate that IL-10-producing B-cells play a crucial role in regulating MG activation, proinflammatory cytokine release and M2 phenotype induction, post-MCAO, with heightened sensitivity of female MG to IL-4 and IL-10. This study, coupled with our previous demonstration of increased numbers of transferred IL-10+ B-cells in the ischemic hemisphere, provide a mechanistic basis for local regulation by secreted IL-10 and IL-4 as well as direct B-cell/MG interactions that promote M2-MG.
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Abbreviations
- BBB:
-
Blood brain barrier
- CNS:
-
Central nervous system
- MCAO:
-
Middle cerebral artery occlusion
- WT:
-
Wild-type
- TNF-α:
-
Tumor necrosis factor α
- CD:
-
Cluster of differentiation
- MHC II:
-
Major histocompatibility complex II
- RPMI:
-
Roswell Park Memorial Institute
- IL:
-
Interleukin
- FACS:
-
Fluorescence activated cell sorter
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Acknowledgments
The authors wish to thank Gail Kent for assistance with manuscript submission. This work was supported by National Institutes of Health/National Institute of Neurological Disorders and Stroke grant 1RO1 NS075887. This material is based upon work supported in part by the Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development, Biomedical Laboratory Research and Development. The contents do not represent the views of the Department of Veterans Affairs or the United States Government.
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The authors declare that they have no conflict of interest.
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All applicable international, national and institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the Institutional Animal Committee.
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Sheetal Bodhankar and Andrew Lapato contributed equally to this work.
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Bodhankar, S., Lapato, A., Chen, Y. et al. Role for microglia in sex differences after ischemic stroke: importance of M2. Metab Brain Dis 30, 1515–1529 (2015). https://doi.org/10.1007/s11011-015-9714-9
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DOI: https://doi.org/10.1007/s11011-015-9714-9