Multiple sclerosis–associated CD226 variant disrupts vitamin D–induced type I Treg differentiation by impairing LFA-1 activation
Saniya Kari, Aymeline Debonlier, Thibault Angles, Beatriz Chaves, Charles Grosjean, Florence Bucciarelli, Valérie Duplan-Eche, Lucie Nozeran, Jessica Lavery, Elena Morandi, Beatrice Pignolet, Max Mimpen, Joost Smolders, Roland Liblau, Abdelhadi Saoudi, Jan Damoiseaux, Frederick Masson, Loïc Dupré, Anne L. Astier
Saniya Kari, Aymeline Debonlier, Thibault Angles, Beatriz Chaves, Charles Grosjean, Florence Bucciarelli, Valérie Duplan-Eche, Lucie Nozeran, Jessica Lavery, Elena Morandi, Beatrice Pignolet, Max Mimpen, Joost Smolders, Roland Liblau, Abdelhadi Saoudi, Jan Damoiseaux, Frederick Masson, Loïc Dupré, Anne L. Astier
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Research Article Autoimmunity Immunology Inflammation

Multiple sclerosis–associated CD226 variant disrupts vitamin D–induced type I Treg differentiation by impairing LFA-1 activation

Abstract

Multiple sclerosis (MS) is a complex inflammatory disease of the CNS resulting from an intricate interplay between genetic predisposition and environmental factors. Vitamin D (VD) deficiency is one of the established risk factors for MS. CD46 costimulation of CD4+ T cells induces a switch from Th1 to type I regulatory cells (Tr1), characterized by increased IL-10 production. This switch is impaired in MS T cells but can be restored by VD, which also strongly promotes expression of CD226 on CD46-activated T cells. The rs763361 polymorphism in the CD226 gene, resulting in a nonsynonymous Gly307Ser variant, is associated with increased risk for MS. Herein, we show that expression of this CD226 risk allele disrupts the ability of CD46-activated T cells to operate the IFN-γ/IL-10 switch upon VD exposure. Mechanistically, the risk variant impairs activation of the integrin LFA-1, which promotes the Tr1 phenotype. LFA-1–mediated Tr1 differentiation is also impaired in MS T cells expressing the CD226 risk allele upon CD46 and VD stimulation. Our study unveils how, in the context of MS susceptibility, a genetic polymorphism and an environmental factor act in concert to control the differentiation of Tr1 cells.

Authors

Saniya Kari, Aymeline Debonlier, Thibault Angles, Beatriz Chaves, Charles Grosjean, Florence Bucciarelli, Valérie Duplan-Eche, Lucie Nozeran, Jessica Lavery, Elena Morandi, Beatrice Pignolet, Max Mimpen, Joost Smolders, Roland Liblau, Abdelhadi Saoudi, Jan Damoiseaux, Frederick Masson, Loïc Dupré, Anne L. Astier

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Figure 1

CD226 knockout by CRISPR/Cas9 impairs IL-10 production by CD46-activated primary CD4+ T cells.

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CD226 knockout by CRISPR/Cas9 impairs IL-10 production by CD46-activated...
CD4+ T cells from HD homozygous for the CD226 rs763361 C/C genotype were nucleofected with either a control guide (sgCtrl) or 2 different guides directed against CD226 (sgCD226-1 and sgCD226-2), and then activated in vitro by anti-CD3 ± anti-CD46 antibodies. (A) Representative histograms and (B) data for all donors for CD226 expression (n = 2–6 donors according to the amounts of cells recovered). (C) Representative histogram and (D) data for all donors for CD46 and CD69 expression (n = 2–6 donors depending on the amounts of cells recovered). (E and F) Percentage of Tr1 based on CD49b and LAG3 coexpression after 5 days (n = 5 or 6 donors). (G) Representative dot plots and (H) data for all donors for intracellular production of IL-10, IFN-γ, and IL-17 by CD46-activated T cells for 3 days after transfection by sgCtrl or sgCD226-2 (n = 8 for each guide). (I) Secretion of IL-10, IFN-γ, and IL-17A was measured in culture supernatants by CBA in CD46-activated T cells for 5 days after transfection by sgCtrl or sgCD226-2 (n = 12 for each guide). For H and I, normalization was also done by calculating the relative change compared with cells with the sgCtrl (100%). Six independent experiments were performed that included sgCtrl and sgCD226-2. Mean ± SEM are shown. One-way ANOVA with Šídák’s post test (B, D, and F) or Wilcoxon t test (H and I).