Fig. 5
NY-BR-11241-1249 is a natural H2-Db-restricted CTL epitope. HLA-DRB1*0401tg mice were immunized i.p. either with 5 × 108 pfu Ad.NY-BR-1 (n = 3) or with 5 × 108 pfu Ad. Control (n = 3) and splenocytes were harvested 14 days post immunization. a Splenocytes were incubated overnight with 5 μg/mL of synthetic peptides representing predicted NY-BR-1 epitopes. IFNγ secreting CD8+ T cells were analyzed by IFNγ catch assay and the percentage of CD8+ T cells secreting IFNγ is depicted. Immunization with Ad.NY-BR-1 resulted in CD8+ T cells reactive against peptides NY-BR-11241-1249. b and c Splenocytes from immunized mice were stained with fluorescently labelled H2-Db dextramers loaded with NY-BR-11241-1249 (NY-BR-1-specific dextramer) or HPV 16 E749–57 (control dextramer). b Proportion of dextramer+CD8+ T cells among viable CD14−CD3+ splenocytes from control mice (left panel) or from an Ad.NY-BR-1 immunized mice (right panel). c Representative dot plot depicting the proportion of dextramer+CD8+ T cells among viable CD14−CD3+ splenocytes from a control mouse (left panel) or from an Ad.NY-BR-1 immunized mouse (right panel)