Figure 4.

Myelin-specific T-reg accumulate in the CNS. Mononuclear cells were prepared from the CNS of MOG/CFA-immunized mice at different stages of EAE as indicated. (a) CD4⁺ T cells stained with MOG_35–55/IAb tetramer or a control-tetramer. The numbers indicate percentages within the CD4⁺ T-cell gate. (b) Absolute numbers of MOG tetramer–positive T-reg versus T-eff recovered from the spleen (SPL) and the CNS before disease onset (Presympt), at the peak of disease and during recovery. The ratio of MOG-specific T-reg:T-eff is indicated for each disease phase. (c) Comparison of CCR5 expression on CD4⁺Foxp3/GFP⁺ T cells (T-reg) and CD4⁺Foxp3/GFP⁻ T cells (T-eff) isolated from the lymph nodes (LN), spleen (SPL) and CNS of EAE mice at the peak of disease (day 14). Mean (+ s.d.) of isotype-controlled expression of CCR5. n = 3. *P < 0.004, t-test. (d) CD103 (integrin αEβ7) expression on T-reg (CD4⁺Foxp3/GFP⁺) versus T-eff (CD4⁺Foxp3/GFP⁻) at the peak of disease. Mean (+ s.d.) of isotype-controlled expression of CD103. n = 4–7. *P < 0.003, **P < 0.002, t-test. (e) CD103⁺ cells are enriched in MOG tetramer–positive T-reg. Three-color staining for CD4, MOG tetramer and CD103 in CNS mononuclear cells from EAE mice. The fraction (as a percentage) of CD103⁺ cells (black line) versus control IgG (shaded curve) is shown in the MOG tetramer–negative and MOG tetramer–positive T-reg (top) and T-eff (bottom) populations.