Known concentrations of the purified mouse IgE myeloma protein, provided by the manufacturer, were used to generate a standard curve to convert OD readings of samples to ng/mL. Sensitivity of assays was 3–4 ng/mL. Data from experiments were reported as mean ± SEM. Mean values of normally distributed data were compared using the one-way or two-way Analysis of Variance (anova) and P-values were assigned using Regorafenib purchase Tukey post hoc analysis or two-way anova followed by Bonferroni post-test, as depicted in each figure. Differences of P < 0·05 were considered significant. Statistical tests were performed

using the GraphPad Prism Software. Primary infection of mice with S. venezuelensis VX-809 resulted in egg elimination in faeces after 7 days of infection, confirming the success of the infection procedure (results not shown), except for mice infected

with one infective larva (very low-dose group, L1), in which only four of 10 mice eliminated parasite eggs in faeces. Upon a challenge infection, there was no difference in the number of adult worms recovered from the small intestine (Figure 2a), eggs eliminated in faeces (Figure 2b) or female fecundity index (Figure 2c) in mice that were initially infected with one larva (L1) compared with mice that were primary infected (L0). In contrast, mice previously infected with 10 (low-dose group, L10), 100 (normal-dose group, L100) or 500 (high-dose group, L500) parasite larvae had a significant reduction in the number of adult worms recovered

from OSBPL9 the small intestine (Figure 2a), eggs eliminated in faeces (Figure 2b) and female fecundity (Figure 2c) after 7 days of challenge when compared with primary infected animals. As L1 group did not show protection against challenge infection and L100 and L500 groups had similar worm elimination profile during challenge, the following analyses were comparatively carried out between primary infected (L0 group), low-dose exposed animals (L10 group) and high-dose exposed animals (L500). Even though there were no significant differences in worm burden, egg production or fecundity after the challenge infection between L10 and L500 groups; it must be highlighted that no adult worms were recovered from the small intestine and no eggs were encountered in the faeces amongst the animals from the L500 group, suggesting that high-dose priming group was able to completely abolish challenge infection before adult worm maturation. In contrast, in the low-dose priming group, adult worms in the intestine as well as eggs in the faeces were detected in most of the challenged animals (Figure 2a, b). The number of larvae in the lungs was assessed to verify whether parasite reduction was also detected early in the course of infection.