FIGURE 5.

Specific OVA peptide may increase N1ICD, perforin and granzyme B expression in CD8⁺ T cells. To study the effects of OVA-specific Ag stimulation on N1ICD expression in the presence of GSI, OVA-specific CD8⁺ T cells were prepared, as described in Materials and Methods. Briefly, C57BL/6 mice were immunized with PBS, as a negative control, with the parental EL4 cell line as a mock control, or with E.G7-OVA cells, engineered to express an OVA peptide. After 16 days splenocytes were harvested from these mice, pretreated in vitro with GSI then stimulated with the OVA-specific E.G7 cells for 5 days. N1ICD expression was determined by immunoblotting (A) and intracellular staining (B). Dotted or open historams represent the conjugated isotype control. C, Together, intracellular staining and flow cytometric analysis were used to assess the percentage of CD8⁺ T cells expressing perforin and granzyme B. Data shown are representative of three individual replicates.