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The pathogenic relevance of αM-integrin in Guillain–Barré syndrome

Abstract

The molecular determinants and mechanisms involved in leukocyte trafficking across the blood–nerve barrier (BNB) in the acute inflammatory demyelinating polyradiculoneuropathy (AIDP) variant of Guillain–Barré syndrome are incompletely understood. Prior work using a flow-dependent in vitro human BNB model demonstrated a crucial role for αM-integrin (CD11b)-intercellular adhesion molecule-1 interactions in AIDP patient leukocyte trafficking. The aim of this study is to directly investigate the biological relevance of CD11b in AIDP pathogenesis. Immunohistochemistry was performed on three AIDP patient sural nerve biopsies to evaluate endoneurial leukocyte CD11b expression. A severe murine experimental autoimmune neuritis (sm-EAN) model was utilized to determine the functional role of CD11b in leukocyte trafficking in vivo and determine its effect on neurobehavioral measures of disease severity, electrophysiological assessments of axonal integrity and myelination and histopathological measures of peripheral nerve inflammatory demyelination. Time-lapse video microscopy and electron microscopy were employed to observe structural alterations at the BNB during AIDP patient leukocyte trafficking in vitro and in situ, respectively. Large clusters of endoneurial CD11b+ leukocytes associated with demyelinating axons were observed in AIDP patient sural nerves. Leukocyte CD11b expression was upregulated during sm-EAN. 5 mg/kg of a function-neutralizing monoclonal rat anti-mouse CD11b antibody administered after sm-EAN disease onset significantly ameliorated disease severity, as well as electrophysiological and histopathological parameters of inflammatory demyelination compared to vehicle- and isotype antibody-treated mice. Consistent with in vitro observations of leukocyte trafficking at the BNB, electron micrographs of AIDP patient sural nerves demonstrated intact electron-dense endoneurial microvascular intercellular junctions during paracellular mononuclear leukocyte transmigration. Our data support a crucial pathogenic role of CD11b in AIDP leukocyte trafficking, providing a potential therapeutic target for demyelinating variants of Guillain–Barré syndrome.

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Acknowledgments

Special thanks to past and current employees of the Shin J. Oh Muscle and Nerve Histopathology Laboratory, University of Alabama at Birmingham for archived peripheral nerves and processed slides from AIDP patients and controls. Special thanks to EMLABS Inc. for performing the electron microscopy work. This project was supported by National Institutes of Health (NIH) Grant R21 NS078226 (2012-2015) and institutional funds from the University of Alabama at Birmingham. The funding sources had no involvement in the conduct of the research, manuscript preparation, data collection/analyses or decision to submit this work for publication. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

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Correspondence to Eroboghene E. Ubogu.

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Conflict of interest

E.E.U. has received royalties from Baylor Licensing Group for simian virus-40 large T-antigen immortalized human endoneurial endothelial cells and from Springer Science + Business Media for an edited book on laboratory protocols that describes the flow-dependent in vitro BNB assay. C.D., S.P.P. and E.S.H. have nothing to disclose.

Electronic supplementary material

401_2016_1599_MOESM1_ESM.mp4

Supplementary Video 1. AIDP patient leukocyte trafficking at the in vitro bloodnerve barrier. A compressed two minute video generated via real-time time-lapse digital microscopy demonstrates leukocytes from an untreated AIDP patient (phase bright) rolling and undergoing arrest and firm adhesion between cytokine stimulated confluent BNB-forming endoneurial endothelial cells (dark spindle-shaped background cells) under a continuous flow velocity of 1 mm/s in vitro. Diapedesis, detected by a change in leukocyte phase from bright to dark, was observed via the paracellular route with slight endothelial cell displacement, similar to in situ ultrastructural observations in AIDP patient sural nerve biopsies. Leukocytes were attracted to sites of firm adhesion and active transmigration, consistent with inflammatory cell clusters seen in AIDP patient biopsies. Frame rate = 10 frames per second (10X normal, compressing 20 minutes in real time) (MP4 7568 kb)

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Dong, C., Palladino, S.P., Helton, E.S. et al. The pathogenic relevance of αM-integrin in Guillain–Barré syndrome. Acta Neuropathol 132, 739–752 (2016). https://doi.org/10.1007/s00401-016-1599-0

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Keywords

  • Acute inflammatory demyelinating polyradiculoneuropathy
  • Blood–nerve barrier
  • CD11b
  • Experimental autoimmune neuritis
  • Guillain–Barré syndrome
  • Integrin