Because the factor ‘age’ has three levels (1, 6 and 20 weeks), post hoc testing was performed in case of significant main effects of age. When significant interaction effects were found, these instead of significant main effects were evaluated statistically by post hoc analyses. Outcomes of post hoc tests are shown on the figures. For clarity, only significant and relevant comparisons are presented,

for example, the 0.1-μg dose in 1-week-old mice is compared to the 0.1-μg dose, but not to the 10-μg dose, in older mice. The limit for statistical significance was set to P < 0.05. To investigate how sex, age and dose influenced sensitization and allergic inflammation in a standard airway allergy mouse model, female and male mice of

different age groups were sensitized and boosted i.p. with different learn more doses of OVA and challenged with three i.n. instillations of OVA. Significant main and interaction effects are given in Table 2, and results Temsirolimus of the post hoc tests are displayed on the figures. OVA-specific IgE and IgG1 were measured in serum both before and after the airway challenges with OVA and statistical analyses revealed that dose, age and sex affected the antibody response in a similar way both before and after OVA challenges. This implies that the relationship between the groups was equivalent, and, therefore, only the antibody levels after allergen challenge are shown. Following the airway challenges, a significant interaction of sex, allergen dose and age was found for the OVA-specific IgE response (Table 2). For clarity, females and males are depicted in separate PIK3C2G graphs (Fig. 1A, B). Overall, the IgE response in 1-week-old mice differed from the responses of older age groups. One-week-old females responded with significantly higher IgE production to sensitization with the 10-μg dose compared to the 0.1- and 0-μg dose (Fig. 1A). A comparable relationship

was observed for the 1-week-old males (Fig. 1B). The effect of dose was reversed in the older females with the highest IgE levels found following immunization with 0.1 μg OVA. The effect of the 0.1 and 10 μg doses did not differ in male mice (Fig. 1A, B). In 1-week-old mice, no effect of sex could be observed. After immunization with 0.1 μg OVA, the mean IgE response in 6- and 20-week-old females was higher compared with the males, but only statistically significant for 6-week-old mice (‘S’ in Fig. 1A, B). A significant effect of age on IgE production was only seen in female mice. At 6 and 20 weeks of age, females responded with significantly higher IgE levels to the 0.1-μg dose compared to 1-week-old females (* in Fig. 1A). No differences in IgE levels were observed between the oldest age groups. Interestingly, no effect of sex was seen on OVA-specific IgG1 production, and both sexes are therefore combined in Fig. 1C. A significant dose and age interaction effect was found (Table 2).