mDC-independent LNDC/CD4⁺ T cell activation. Ear resection in the vG model 30 min after vaccination. (a) Percentage of CD8a⁺ or CD11b^hi cDCs in the ALN 24 h after ear vaccination. PBS, WT, and vG groups are displayed. ANOVA (light gray), P < 0.0001; ANOVA (dark gray), P < 0.0001 (n = 4). *, P < 0.05; ***, P < 0.001. (b) CFSE-labeled OT-II T cells were transferred into C57BL/6 recipients. Recipients were vaccinated in the ear and separated into WT and vG groups. ALNs were isolated at 60 h after injection and analyzed by flow cytometry. (left) Percentage of OT-II T cells by division number as assessed by CFSE dilution. ANOVA, P > 0.1. (right) Representative plot of CFSE dilution in WT and vG groups (n = 4 mice/group). (c and d) Adoptive transfers were established as in c. OT-II T cell expression (MFI) of CXCR3 (c) and CD40L (d) was assessed by T cell division to track acquisition over time. (c) Two-way ANOVA, P < 0.0001 overall, P < 0.05 between groups. (d) Two-way ANOVA, P < 0.0001 overall, P > 0.1 between groups (n = 4 mice/group). Mean ± SEM.