Figure 3.

The occurrence of new reactivity of 2D2 T cells depends on MHC II. 2D2 CD4⁺ T cells were sorted from spleen of 2D2 mice without central nervous system autoimmune disease, and transferred intravenous (i.v.) into Rag2^−/− or Rag2^−/−MHC II^−/− recipient mice before 60 min of middle cerebral artery occlusion (MCAO) surgery. A, Schematic graph showing the experimental design. B, At day 4 or day 7 after reperfusion, 2D2 CD4⁺ T cells were reisolated from brains of Rag2^−/− or Rag2^−/−MHC II^−/− mice and then stained with PE-labeled tetramers containing MOG (myelin oligodendrocyte glycoprotein)_35-55, MOG_91-108, or MOG_103-125 epitopes. Flow cytometry plots show MOG epitope-specific 2D2 CD4⁺ T cells. All gates were set using fluorescence minus one controls. C, Bar graph shows quantification of surface expression of antigen-specific T-cell receptors at day 4 and day 7 after reperfusion. n=15 per group. Error bars represent SEM. *P<0.05, **P<0.01.