Patients With Severe Multiple Sclerosis Exhibit Functionally Altered CD8 ⁺ Regulatory T Cells

<h3>Background and Objectives</h3> Multiple sclerosis (MS) is a chronic inflammatory and demyelinating disease of the CNS. Studies of immune dysfunction in MS have mostly focused on CD4⁺ Tregs, but the role of CD8⁺ Tregs remains largely unexplored. We previously evidenced the suppressive properties of rat and human CD8⁺CD45RC^low/neg Tregs from healthy individuals, expressing Forkhead box P3 (FOXP3) and acting through interferon-gamma (IFN-γ), transforming growth factor beta (TGFβ), and interleukin-34 (IL-34). secretions to regulate immune responses and control diseases such as transplant rejection. To better understand CD8⁺CD45RC^low/neg Tregs contribution to MS pathology, we further investigated their phenotype, function, and transcriptome in patients with MS. <h3>Methods</h3> We enrolled adults with relapsing-remitting MS and age-matched and sex-matched healthy volunteers (HVs). CD8⁺ T cells were segregated based on low or lack of expression of CD45RC. First, the frequency in CSF and blood, phenotype, transcriptome, and function of CD8⁺CD45RC^low and ^neg were investigated according to exacerbation status and secondarily, according to clinical severity based on the MS severity score (MSSS) in patients with nonexacerbating MS. We then induced active MOG_35-55 EAE in C57Bl/6 mice and performed adoptive transfer of fresh and expanded CD8⁺CD45RC^neg Tregs to assess their ability to mitigate neuroinflammation in vivo. <h3>Results</h3> Thirty-one untreated patients with relapsing-remitting MS were compared with 40 age-matched and sex-matched HVs. We demonstrated no difference of CSF CD8⁺CD45RC^low and CD8⁺CD45RC^neg proportions, but blood CD8⁺CD45RC^low frequency was lower in patients with MS exacerbation when compared with that in HVs. CD8⁺CD45RC^neg Tregs but not CD8⁺CD45RC^low showed higher suppressive capacities in vitro in MS patients with exacerbation than in patients without acute inflammatory attack. In vitro functional assays showed a compromised suppression capacity of CD8⁺CD45RC^low Tregs in patients with nonexacerbating severe MS, defined by the MSSS. We then characterized murine CD8⁺CD45RC^neg Tregs and demonstrated the potential of CD45RC^neg cells to migrate to the CNS and mitigate experimental autoimmune encephalomyelitis in vivo. <h3>Discussion</h3> Altogether, these results suggest a defect in the number and function of CD8⁺CD45RC^low Tregs during MS relapse and an association of CD8⁺CD45RC^low Tregs dysfunction with MS severity. Thus, CD8⁺CD45RC^low/neg T cells might bring new insights into the pathophysiology and new therapeutic approaches of MS.