Fig 2: Quantification of immune cells in the ventral surface of the tongue, the mouth floor and the cheek from rats, dogs, minipigs and monkeys.Mucosal tissues from animal species were processed for immunohistology analysis, as described in Methods. Cell counting was performed on slides labeled with Abs specific for APC (anti-MHC-II, anti-CD163, anti-CD172a) and T cell (anti-CD3 and anti-CD4) markers or stained with toluidine blue for mast cells to evaluate the mean number of positive cells per field using a light microscope (magnification x400). All areas (epithelium (Epith.), Lamina propria (LP) and muscle) were scored. Histograms represent the mean + SEM with n = 3.

Fig 3: Abundance of T regulatory cells in lung tissue. A significant comparable increase in FoxP3+:CD3+ cells ratio was induced by d2-OID and VD3 treatments (p < 0.05 vs sham); the combination treatment d2-OID + VD3 was even more efficacious in inducing such an increase (p < 0.001 vs sham, p < 0.01 vs d2-OID and VD3 treatments) Values are expressed as mean ± SD (n = 7). ***p < 0.001 **p < 0.01. *p < 0.05

Fig 4: Relative frequency of peripheral blood lymphocyte subsets in IL-15-deficient and IL-15-proficient, control and vaccinated NeuT mice. Lymphocyte subsets were analyzed by flow cytometry in control and vaccinated mice 20 hours after vaccination. A lymphocyte gate was defined in the FSC-SSC dot plot then the percentage of B220+, CD3+, CD4+, CD8+ and Nkp46 and DX5 double-positive cells was determined. Significance of comparisons (Student’s t test): * p <0.05, IL15KO/NeuT versus NeuT mice; # p <0.05, vaccinated versus control mice within the same strain. Mean ± SEM is shown (three to five mice per group). IL15KO/NeuT BALB/c mice transgenic for rat HER2/neu oncogene and knocked out for IL-15, NeuT BALB/c mice transgenic for rat HER2/neu oncogene, SEM standard error of the mean

Fig 5: The presence of immune cells during histoculture(A) Visualization of CD3, CD8 and FoxP3 T-cell staining at day 7. In the merged image, one can distinguish between T helper cells (CD3+), cytotoxic T cells (CD3+, CD8+) and regulatory T cells (CD3+, FoxP3+). (B) Visualization of CD68 and CD163 macrophage staining at day 0. In the merged image, one can distinguish M2 macrophages (CD68/CD163).