For the 0.1-μg dose, lymphocyte and eosinophil numbers were significantly higher in 20- compared with 1-week-old mice (* in Fig. 3A, B). For the 10-μg dose, this was opposite; the cell numbers decreased with age (Fig. 3A, B). In a separate study, mice were sensitized by i.n. instillation of OVA in Al(OH)3 and challenged i.n. with OVA. The main and interaction effects are reported above the figures. When a significant effect of age or a significant sex and age interaction

was found, the result of the post hoc test is given on the figure. Fig. 4A shows the OVA-specific IgE response in 1-, 6- and 20-week-old female and male mice. Significant main effects of both sex and age were found. Sensitized females produced higher levels of OVA-specific IgE compared with males (Fig. 4A). GS1101 Further, the IgE response increased with age as 20-week-old mice had significantly higher levels than 1-week-old mice. The same pattern was observed for OVA-specific IgG1 production; females had significantly higher antibody production than males, and the response in both sexes increased with age (Fig. 4B). Cells from both SLNs and MLNs were stimulated with OVA ex

vivo. In MLNs, IL-4 was undetectable. Only IL-10 secretion was influenced by the sex of the mice, with females releasing significantly more IL-10 than males (Fig. 5A). IL-5 and IL-13 secretion was higher in 1-week-old mice compared with Selleckchem GSK3 inhibitor older mice (Fig. 5B, C). INFγ was affected by age in the same manner as IL-17A secretion (Fig. 5D, E); 6-week-old mice had significantly lower IFNγ and IL-17A secretion than until 20-week-old mice and for IFNγ also significantly

lower than the 1-week-old mice. A significant age and sex interaction was found for the total number of cells in MLNs (Fig. 5F). The post hoc test revealed that only in the oldest age group did females have significantly higher number of cells compared with males (bracket in Fig. 5F). In SLNs, IL-4, -5, -10, -13 and IFNγ were undetectable and IL-17A produced at very low levels (data not shown and Fig. 5G). IL-17A production increased with age but was not affected by sex. The total number of cells in SLNs was unaffected by both sex and age (Fig. 5H). Control groups of mice were immunized i.n. with OVA alone. When comparing the OVA and OVA + Al(OH)3 treatments, MLN cell numbers, but not SLN cell numbers, were highly increased after using the adjuvant for sensitization, and this was observed for all age groups (data not shown). In contrast to the control groups (data not shown), a pronounced airway inflammatory cell influx dominated by macrophages, lymphocytes, some epithelial cell shedding and in particular by eosinophils was found in BALF of the mice. However, only lymphocytes, epithelial cells and eosinophils were significantly affected by the investigated experimental factors. The number of lymphocytes, eosinophils and epithelial cells in BALF was significantly higher in female mice compared with male mice (Fig. 6A, B, C).