Fig. 4

Evaluation of the efficacy of compound C216 as a prophylactic vaccine in dog. A Experimental design. B Total serum IgG and IgG2b were measured by ELISA, the optical density (OD) was obtained at 450 nm and the means of the ODs ± SD were calculated. C, D, and E T-cell stimulation was evaluated by IFN-γ ELISpot assay at days 27 and 56. Peripheral blood mononuclear cells were collected from dogs and stimulated in triplicate with Vaxiclase (C) or three pools of E7/HPV16 peptides (D and E). For each dog, the number of spot forming cells (SFC) corresponds to the median of triplicates and was corrected with the negative control (DMSO). The mean \(\pm\) SD of the SFC/10⁶ cells was calculated for the three groups of vaccines. D and E For each E7/HPV16 peptide (116-1j, 116-2jc, and 116-2jd), instead of calculating the mean, the values for SFC/10⁶ cells at days 27 and 56 were divided to the value at day 0, to obtain a ratio for each dog. F The fluorescence was acquired with the Z probe at 488 nm using the Cellvizio imaging system and was corrected with that observed at the non-injected (NI) point, for every dog at days 54 or 55. The corrected fluorescence was calculated according to a ratio between the mean of the fluorescence intensity at the lentiviral point and the mean of the fluorescence intensity of the NI point. Finally, the means of the ratios ± SD were calculated. G ZsGreen mRNA expression was evaluated by RT-ddPCR for the three groups