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Original Contribution |

Enriched CD161high CCR6+ γδ T Cells in the Cerebrospinal Fluid of Patients With Multiple Sclerosis

Lucas Schirmer, MD; Veit Rothhammer, MD; Bernhard Hemmer, MD; Thomas Korn, MD
JAMA Neurol. 2013;70(3):345-351. doi:10.1001/2013.jamaneurol.409.
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Objective  To investigate the expression of CD161 (KLRB1) and CCR6 on human γδ T cells in blood and cerebrospinal fluid (CSF) of patients with a clinically isolated syndrome (CIS) and multiple sclerosis (MS) in relapse.

Design  Flow cytometry analysis of CD161 and CCR6 expression and intracellular cytokine staining for interleukin 17 and interferon-γ on human γδ T cells in blood and CSF samples.

Setting  Department of Neurology, Klinikum rechts der Isar, Technische Universität München, a tertiary referral center.

Patients  Twenty-six patients with CIS/MS in active relapse, 10 patients with other autoimmune disorders, 12 patients with neuroinfectious diseases, and 15 patients with noninflammatory neurological diseases.

Main Outcome Measures  Frequencies of CD161high and CCR6+ γδ T cells in blood and CSF samples of patients with CIS/MS in relapse and control patients.

Results  γδ T cells were increased in both blood and CSF of patients with CIS/MS in relapse as compared with controls with noninflammatory disease. The fraction of CD161high CCR6+ γδ T cells was significantly higher in the CSF of patients with CIS/MS in relapse than of those with systemic autoimmune disorders or controls with noninflammatory disease. The CD161high CCR6+ double-positive γδ T-cell population was further enriched in the CSF in relation to blood in patients with CIS/MS in relapse but not in patients with infectious disease or the other control groups. The CD161high CCR6+ γδ T-cell population was characterized by its capacity to produce interleukin 17.

Conclusion  Interleukin 17–producing CD161high CCR6+ γδ T cells might contribute to the compartmentalized inflammatory process in the central nervous system of patients with MS.

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Figures

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Grahic Jump Location

Figure 1. γδ T cells and γδ T-cell subsets in the blood and cerebrospinal fluid (CSF) of various populations of patients. A, Contour plots with parent gate on CD3+ T cells. γδ T-cell frequencies (in percentage) are shown in the peripheral blood and CSF of patients with clinically isolated syndrome/multiple sclerosis in relapse (CIS/MS in relapse) as compared with control groups (Kruskal-Wallis followed by a Dunn post hoc test). TCR indicates T-cell receptor. B, Contour plots with parent gate on γδ T cells. CD161high CCR6+ γδ T-cell frequencies (in percentage) in blood and CSF samples of patients with CIS/MS in relapse and control patient groups are plotted (Kruskal-Wallis followed by a Dunn post hoc test). Significant differences between groups are marked by a capped line.

Place holder to copy figure label and caption
Grahic Jump Location

Figure 2. Cerebrospinal fluid (CSF) accumulation and functional phenotype of CD161high CCR6+ γδ T cells. A, Histograms with parent gate on γδ T cells. Differential expression of CCR6 (upper panel) and CD161 (lower panel) on γδ T cells in peripheral blood and CSF. B, Scatterplots showing differences in CD161high CCR6+ γδ T-cell frequencies between blood and CSF in patients with clinically isolated syndrome/multiple sclerosis in relapse (CIS/MS in relapse) and control patient groups (Wilcoxon signed rank test). (C) Contour and dot plots with parent gate on γδ T cells (contour plot) and subpopulations of γδ T cells according to their CD161 and CCR6 status (lower panel). Note that interleukin 17 (IL-17)–producing γδ T cells are exclusively found in the CD161high CCR6+ subpopulation. All numbers are given as a percentage of parent population (parts A and C), and in dot plots (part C), numbers are given as mean (SD). Three independent experiments.

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