Expression of MC and Ag in cutaneous atypical APCs contribute to adjuvant-enhanced EP vaccine-mediate Ag-specific CD8⁺ T cell priming and is partly CD8α⁺/CD103⁺ DC-independent.

<p>C57BL/6 mice were vaccinated on days 0 and 21 with 25 μg pDNA by EP, rim was administered 1.25 mg/kg IP the day following each vaccination in MC.OVA.miR142T + rim-treated mice. <b>(A)</b> Splenocytes were extracted 7 days after the final vaccination (day 28) and restimulated with SIINFEKL peptide overnight. IFN-γ SFCs were quantitated by ELISpot. n = 5, *p<0.05, One-way ANOVA, Tukey correction for multiple comparisons. <b>(B)</b> C57BL/6 mice were injected in alternating flanks 21 days apart with 1 x 10⁵ MPEK cells stables (neg. control, OVA, MC.OVA). Rimiducid was administered at 1.25 mg/kg IP 24 hours after each injection of cells. 7 days following the second injection, splenocytes were analyzed for SIINFEKL-specific IFN-γ-secreting CD8⁺ T cell by ELISpot. Error bars represent 95% confidence interval. *p<0.05 when compared to MPEK-OVA, One-way ANOVA, Dunnett’s correction for multiple comparisons. <b>(C)</b> Either C57BL/6 or <i>Batf3</i>^-/- mice were vaccinated in the same manner as described for panel A. SIINFEKL-specific IFN-γ-secreting splenocytes were quantitated by ELISpot. n = 4–5, *p<0.05, One-way ANOVA, Tukey correction for multiple comparisons within mouse strain.