Fig. 3.

Central nervous system (CNS)–infiltrating CD8+ T cells recognize the immunodominant D^b:VP2_121–130 epitope as determined by our Cos-7 co-transfection system. (A) Enzyme-linked immunoabsorbent assay was used to detect interferon (IFN)–γ expression by central nervous systesm (CNS)–infiltrating CD8+ T cells co-cultured with Cos-7 cells co-transfected with D^b plus pcDNA 3.1 His A/VP2. D^b plus empty pcDNA 3.1 His A vector resulted in no IFN-γ expression. In (B), D^b:VP2_121–130 tetramer staining demonstrated that at 7 days postinfection, approximately 55% of all brain-infiltrating CD8+ T cells were specific for the VP2_121–130 epitope confirming the results in (A). These data demonstrate that VP2_121–130 peptide was successfully expressed, processed, and presented by D^b class I molecule demonstrating the feasability of our co-transfection molecular approach to stimulate the response of ex vivo isolated CD8+ T cells. Values presented in (A) are the mean of two wells.