Figure 4.

Antitumor response and in vivo efficacy of SIINFEKL mutated analogs. (A) B16OVA cells were injected into both flanks of female C57BL6/J mice (2.5 × 10⁵ cells/flanks). Mice (eight per group) were treated with PBS (Mock), Ad5D24-CpG human adenovirus (Adjuvant), and either SIINFEKL, SIINFAKL or SIIWFEKL peptides complexed to the virus-adjuvant. Intra-tumor injections were made on day 9, 11, 13 as represented by the asterisks on the x axis. Tumor volumes were measured every 2–3 d by a digital caliper. Tumor volumes normalized against the values on the 9th day are presented as the mean ± SEM; significance was calculated by Two-way ANOVA with Tukey's post hoc test. Spleens, tumors and draining lymph nodes were collected to determine to percentage of (B) total CD8⁺ T-cells (CD19⁻CD3⁺CD8⁺) or (C) SIINFEKL-specific T-cells (double positive Pentamer⁺CD8⁺ gated on CD19⁻CD3⁺). (D) A correlation analysis was performed by plotting for each mouse the % of antigen-specific T-cells against the intensity of the signal (gMFI) of such population. Data from spleens (empty triangles), draining lymph nodes (empty squares) and tumors (black dots) are presented. Log transformed values were used to compute the Pearson's correlation coefficiencies; p-value for the correlation is 0.0002.