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Immune Characterization in Aneurysmal Subarachnoid Hemorrhage Reveals Distinct Monocytic Activation and Chemokine Patterns

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Abstract

The pathophysiology of delayed cerebral ischemia (DCI) after aneurysmal subarachnoid hemorrhage (aSAH) is incompletely understood. Intrathecal activation of inflammatory immune cells is suspected to play a major role for the induction of DCI. The aim of this study is to identify immune cell subsets and mediators involved in the pathogenesis of DCI. We prospectively collected blood and CSF from 25 patients with aSAH at early and late time points. We performed multicolor flow cytometry of peripheral blood and CSF, analyzing immune cell activation and pro-inflammatory cyto- and chemokines. In addition to the primary immune analysis, we retrospectively analyzed immune cell dynamics in the CSF of all our SAH patients. Our results show an increased monocyte infiltration secondary to aneurysm rupture in patients with DCI. Infiltrating monocytes are defined by a non-classical (CD14dim CD16+) phenotype at early stages. The infiltration is most likely triggered by the intrathecal immune activation. Here, high levels of pro-inflammatory chemokines, such as CXCL1, CXCL9, CXCL10, and CXCL11, are detected. The intrathecal cellular activation profile of monocytes was defined by upregulation of CD163 and CD86 on monocytes and a presumable later differentiation into antigen-presenting plasmacytoid dendritic cells (pDCs) and hemosiderophages. Peripheral immune activation was reflected by CD69 upregulation on T cells. Analysis of DCI prevalence, Hunt and Hess grade, and clinical outcome correlated with the degree of immune activation. We demonstrate that monocytes and T cells are activated intrathecally after aSAH and mediate a local inflammatory response which is presumably driven by chemokines. Our data shows that the distinct pattern of immune activation correlates with the prevalence of DCI, indicating a pathophysiological connection to the incidence of vasospasm.

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Abbreviations

aSAH:

aneurysmal subarachnoid hemorrhage

CD:

cluster of differentiation

CSF:

cerebrospinal fluid

DCI:

delayed cerebral ischemia

H&H:

Hunt and Hess grading

IL:

interleukin

mDC:

myeloid dendritic cell

mRS:

modified Rankin scale

PBMCs:

peripheral blood mononuclear cells

pDC:

plasmacytoid dendritic cell

SAH:

subarachnoid hemorrhage

Treg :

regulatory T cells

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Acknowledgments

We thank all patients and relatives for their support and participation in this study. We thank Sabine Wuttke for her excellent graphical support. Our study would not be possible without the excellent technical support of Manuela Kolster, Mareike Holz, Katharina Kolbe, Svenja Zapf, Sarah Matulla, and the FACS Core Facility at UKE.

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Author notes

  1. Malte Mohme, Thomas Sauvigny, Eva Tolosa and Patrick Czorlich contributed equally to this work.

Authors and Affiliations

  1. Department of Neurosurgery, University Medical Center Hamburg-Eppendorf, Martinistr. 52, 20246, Hamburg, Germany

    Malte Mohme, Thomas Sauvigny, Marius Marc-Daniel Mader, Cecile L. Maire, Alessandra Rünger, Franz Ricklefs, Jan Regelsberger, Nils Ole Schmidt, Manfred Westphal, Katrin Lamszus & Patrick Czorlich

  2. Department of Neurology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

    Nils Schweingruber

  3. Institute of Immunology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

    Eva Tolosa

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  1. Malte Mohme
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Correspondence to Malte Mohme.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee (Hamburger Ärztekammer, PV5106) and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Mohme, M., Sauvigny, T., Mader, M.MD. et al. Immune Characterization in Aneurysmal Subarachnoid Hemorrhage Reveals Distinct Monocytic Activation and Chemokine Patterns. Transl. Stroke Res. 11, 1348–1361 (2020). https://doi.org/10.1007/s12975-019-00764-1

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