Fig. 4.

Spleen derived CD8+ T cells are restricted to the epitope derived from the same Theiler’s murine encephalomyelitis virus (TMEV) genomic segment as central nervous system (CNS)–infiltrating CD8+ T cells. (A) Lymphocytes isolated from the spleen of 7-day TMEV-infected mice were cultured for 4 hours with VP2_121–130 peptide and assessed for interferon (IFN)–γ expression with intracellular cytokine staining. No IFN-γ–positive cells were found in wells stimulated with the E7 control peptide (data not shown). In (B), CD8+ T cells isolated from the spleen were co-cultured with Cos-7 cells co-transfected with D^b and all 15 individual TMEV genomic segments. CD8+ T cells demonstrated specificity for segment 2 of the TMEV genome, which encodes the immunodominant VP2_121–130 peptide, when compared with stimulation with Cos-7 cells co-transfected with D^b and empty pcDNA 3.1 His A vector as predicted by enzyme-linked immunoabsorbent assay IFN-γ detection (***p = 0.0004). All wells in (B) were performed in quadruplicate.