Specific alterations in NKG2D⁺ T lymphocytes in relapsing-remitting and progressive multiple sclerosis patients

Background: T lymphocytes exhibit numerous alterations in relapsing-remitting (RRMS), secondary progressive (SPMS), and primary progressive multiple sclerosis (PPMS). The NKG2D pathway has been involved in MS pathology. NKG2D is a co-activating receptor on subsets of CD4⁺ and most CD8⁺ T lymphocytes. The ligands of NKG2D are expressed at low levels in normal tissues but are elevated in MS postmortem brain tissues compared with controls. Whether the NKG2D pathway shows specific changes in different forms of MS remains unclear.

Methods: We performed unsupervised and supervised flow cytometry analysis to characterize peripheral blood T lymphocytes from RRMS, SPMS, and PPMS patients and healthy controls (HC). We used an in vitro microscopy approach to assess the role of NKG2D in the interactions between human CD8⁺T lymphocytes and human astrocytes.

Results: Specific CD8⁺, CD4⁺, and CD4^-CD8^- T cell populations exhibited altered frequency in MS patients' subgroups. The proportion of NKG2D⁺ T lymphocytes declined with age in PPMS patients but not in RRMS and HC. This reduced percentage of NKG2D⁺ cells was due to lower abundance of γδ and αβ CD4^-CD8^- T lymphocytes in PPMS patients. NKG2D⁺ T lymphocytes were significantly less abundant in RRMS than in HC; this was caused by a decreased frequency of CD4^-CD8^- and CD8⁺ T lymphocytes and was not linked to age. Blocking NKG2D increased the motility of CD8⁺ T lymphocytes co-cultured with astrocytes expressing NKG2D ligand. Moreover, preventing NKG2D from interacting with its ligands increased the proportion of CD8⁺ T lymphocytes exhibiting a kinapse-like behavior characterized by short-term interaction while reducing those displaying a long-lasting synapse-like behavior. These results support that NKG2D participates in the establishment of long-term interactions between activated CD8⁺ T lymphocytes and astrocytes.

Conclusion: Our data demonstrate specific alterations in NKG2D⁺ T lymphocytes in MS patients' subgroups and suggest that NKG2D contributes to the interactions between human CD8⁺ T lymphocytes and human astrocytes.