Figure 1.
Quantification of the cellular infiltrate in the skin of mice after sensitization with different vaccine adjuvants: saponin (SAP; light gray), incomplete Freund’s adjuvant (IFA; medium gray), and monophosphoryl lipid A (MPL; dark gray) at 1, 12, 24, 48, 96, 168, and 336 h after stimulation.
The control group (C; white) was inoculated with 0.9% sterile saline (A). Significant differences (p<0.05) between groups are represented by the letters a, b, c, and d, referring to the C, SAP, IFA, and MPL groups, respectively, and ANOVA following Tukey’s test was employed. The dashed line represents the average number of cell nuclei quantified in histological sections of the skin mouse sensitized with saline. Data presented are the mean±SD from groups of five animals/evaluation time. (B) Representative photomicrographs of the cellular infiltrate at 48 h is shown at the bottom of Figure 1. Skin mice sensitized with saline and the vaccine adjuvants: Saponin (B), IFA (C), MPL (D) at a magnification of 20×; bar = 100 µm.
Figure 2.
Enumeration of the different cell types (neutrophils, macrophages, and lymphocytes) in the inflammatory focus in mouse skin after sensitization with vaccine adjuvants: saponin (SAP; light gray), incomplete Freund’s adjuvant (IFA; medium gray), and monophosphoryl lipid A (MPL; dark gray) at 1, 12, 24, 48, 96, 168, and 336 h after stimulation.
The control group was inoculated with 0.9% sterile saline (C; white). The representative time-lapse graphic demonstrates the major inflammatory cells that migrated to the skin after sensitization: neutrophils (light gray), lymphocytes (medium gray), and macrophages (dark gray). Five mice were used in each group/evaluation time.
Figure 3.
Kinetics of pro-inflammatory and regulatory (TNF-α IL-6, IFN-γ IL-2, IL-17, IL-4, and IL-10) cytokines in the skin of mice after sensitization with vaccine adjuvants: saponin (SAP; gray circle), incomplete Freund’s adjuvant (IFA; gray inverted triangle), and monophosphoryl lipid A (MPL; black triangle) at 1, 12, 24, 48, 96, 168, and 336 h after stimuli.
The control group was inoculated with 0.9% sterile saline (C; white square). Significant differences (p<0.05) between groups are represented by the letters a, b, c and d referring to the C, SAP, IFA, and MPL groups, respectively. Five mice were used in each group/evaluation time.
Figure 4.
Cytokine pattern in the homogenate of mouse skin after sensitization with vaccine adjuvants saponin (SAP; light gray), incomplete Freund’s adjuvant (IFA; medium gray), and monophosphoryl lipid A (MPL; dark gray) at 12, 48, and 168 h after stimulation.
The control group was inoculated with 0.9% sterile saline (C; white). The results are normalized as the ratio between the levels of cytokine (pg/mL) and weight of tissue (mg). Significant differences (p<0.05) between groups are represented by the letters a, b, c and d, referring to the C, SAP, IFA, and MPL groups, respectively, and ANOVA following Tukey’s test was employed. Data presented are the mean±SD from groups of five animals/evaluation time.
Table 1.
Hematological profile of peripheral blood from mice after sensitization with vaccine adjuvants: saponin (SAP), incomplete Freund’s adjuvant (IFA), and monophosphoryl lipid A (MPL) at 1, 12, 24, 48, 96, 168, and 336 h after stimulation.
Figure 5.
Leukocyte immunophenotypic profile in peripheral blood of mice sensitized with vaccine adjuvants: saponin (SAP; light gray), incomplete Freund’s adjuvant (IFA; medium gray), and monophosphoryl lipid A (MPL; dark gray) at 24 or 48 h after stimulation.
The control group was inoculated with 0.9% sterile saline (C; white). The bar graphs present the percentage of cells expressing CD14+ (monocytes), CD19+ (B lymphocytes), and CD4+ and CD8+ (T-lymphocyte subsets). Significant differences (p<0.05) between groups are represented by the letters a, b, c and d, referring to the C, SAP, IFA, and MPL groups, respectively, and ANOVA following Tukey’s test was employed. Data presented are the mean±SD from groups of five animals/evaluation time.