Figure 5.

Chitosan-SRA siRNA complex augment chaperone vaccine-induced antigen-specific CTL response. A-B. DCs were treated with SRA or Scram siRNA complexed with chitosan, followed by pulsing with hsp110-gp100 complex (A) or hsp110-ICD complex (B) for 5 h. DCs were then co-cultured with gp100 or ICD-specific T cells, respectively. ELISA analysis of IFN-γ production and ³H thymidine uptake assays for T cell proliferation were performed. (C) C57BL/6 mice were adoptively transferred with naive Pmel cells on day 0 and immunized on day 1 with chitosan-SRA siRNA complex together with chaperone vaccine. Lymph node cells were analyzed on day 5 for gp100_25-33-reactive, IFN-γ-expressing CD8⁺ or CD90.1⁺ T cells using intracellular cytokine staining and flow cytometry. (D) In vivo CTL assays were performed to assess the cytolytic activity of gp100-specific CD8⁺ T cells against gp100_25–33-pulsed, CFSE-labeled splenocytes following immunotherapy. Percentage of target killing is indicated by the numbers in parentheses. The results are representative of three independent experiments. *p < 0.05; **p < 0.01.