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Regulatory CD8+CD122+ T-cells predominate in CNS after treatment of experimental stroke in male mice with IL-10-secreting B-cells

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Abstract

Clinical stroke induces inflammatory processes leading to cerebral and splenic injury and profound peripheral immunosuppression. IL-10 expression is elevated during major CNS diseases and limits inflammation in the brain. Recent evidence demonstrated that transfer of IL-10+ B-cells reduced infarct volume in male C57BL/6J (wild-type, WT) recipient mice when given 24 h prior to or 4 h after middle cerebral artery occlusion (MCAO). The purpose of this study was to determine if passively transferred IL-10+ B-cells can exert therapeutic and immunoregulatory effects when injected 24 h after MCAO induction in B-cell-sufficient male WT mice. The results demonstrated that IL-10+ B-cell treated mice had significantly reduced infarct volumes in the ipsilateral cortex and hemisphere and improved neurological deficits vs. Vehicle-treated control mice after 60 min occlusion and 96 h of reperfusion. The MCAO-protected B-cell recipient mice had less splenic atrophy and reduced numbers of activated, inflammatory T-cells, decreased infiltration of T-cells and a less inflammatory milieu in the ischemic hemispheres compared with Vehicle-treated control mice. These immunoregulatory changes occurred in concert with the predominant appearance of IL-10-secreting CD8+CD122+ Treg cells in both the spleen and the MCAO-affected brain hemisphere. This study for the first time demonstrates a major neuroprotective role for IL-10+ B-cells in treating MCAO in male WT mice at a time point well beyond the ~4 h tPA treatment window, leading to the generation of a dominant IL-10+CD8+CD122+ Treg population associated with spleen preservation and reduced CNS inflammation.

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Abbreviations

CNS:

Central nervous system

MCAO:

Middle cerebral artery occlusion

WT:

wild-type Green Fluorescent Protein (GFP)

TNF-α:

Tumor necrosis factor α

INF-γ:

Interferon γ

CD:

Cluster of Differentiation

MHC II:

Major Histocompatibility Complex II

RPMI:

Roswell Park Memorial Institute

IL:

Interleukin

PBS:

Phosphate-buffered saline

DNase I:

Deoxyribonuclease I

FACS:

Fluorescence Activated Cell Sorter

PI:

propidium iodide.

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Acknowledgments

The authors wish to thank Gail Kent for the submission of the manuscript. This work was supported by NIH/NINDS 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.

Competing interests

The authors declare no competing financial interests

Authors’ contribution

SB designed and performed the immunology experiments, carried out statistical analyses, prepared graphics and wrote the manuscript; YC performed the MCAO procedures, carried out statistical analyses, prepared the graphics and wrote the methods and results for infarct volume data; AL assisted in tissue preparations and acquisition of immunological data; AAV critiqued and edited the manuscript; SJM and JAS directed study design and data analysis of the MCAO experiments and edited the manuscript; HO directed the overall study, supervised the immunological studies and data analysis and edited the manuscript. All authors read and approved the final version of the manuscript.

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Authors and Affiliations

  1. Neuroimmunology Research, VA Medical Center, Portland, OR, USA

    Sheetal Bodhankar, Andrew Lapato, Arthur A. Vandenbark & Halina Offner

  2. Department of Neurology, Oregon Health & Science University, Portland, OR, USA

    Sheetal Bodhankar, Andrew Lapato, Arthur A. Vandenbark & Halina Offner

  3. Department of Molecular Microbiology & Immunology, Oregon Health & Science University, Portland, OR, USA

    Arthur A. Vandenbark

  4. Department of Anesthesiology & Perioperative Medicine, Oregon Health & Science University, Portland, OR, USA

    Yingxin Chen, Stephanie J. Murphy, Julie A. Saugstad & Halina Offner

  5. Neuroimmunology Research, R&D-31, Portland Veterans Affairs Medical Center, 3710 SW US Veterans Hospital Rd, Portland, OR, 97239, USA

    Arthur A. Vandenbark & Halina Offner

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  1. Sheetal Bodhankar
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  2. Yingxin Chen
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  3. Andrew Lapato
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  7. Halina Offner
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Corresponding author

Correspondence to Halina Offner.

Additional information

Sheetal Bodhankar and Yingxin Chen contributed equally to this work

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Bodhankar, S., Chen, Y., Lapato, A. et al. Regulatory CD8+CD122+ T-cells predominate in CNS after treatment of experimental stroke in male mice with IL-10-secreting B-cells. Metab Brain Dis 30, 911–924 (2015). https://doi.org/10.1007/s11011-014-9639-8

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  • DOI: https://doi.org/10.1007/s11011-014-9639-8

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