Figure 1.
Figure 1. Characterization of DC subset contribution and Tslp induction during Th2 immune responses to Nb and DBP-FITC. LNs were collected from mice immunized with 600 Nb or DBP-FITC and respective controls at the indicated time points. (A) C57BL/6, Langerin-DTR, and Batf3−/− mice were immunized as indicated. DT was injected i.p. 1 d before and 1 d after immunization. On day 7 after immunization, LN cells were collected and processed for intracellular cytokine staining; in vitro restimulation was with PMA and ionomycin (Nb) or anti-CD3 and anti-CD28 (DBP-FITC). Bar graphs show mean + SEM for 4–8 mice/group. Data are from one of two experiments that gave similar results. P-values were determined using one-way ANOVA with the Bonferroni post-test. (B) C57BL/6 mice were immunized as indicated, and dLN were collected for flow cytometric analysis at the indicated times after treatment. DC subsets were defined as in Fig. S1. Bar graphs show the mean + SE of four (Nb) or seven (DBP-FITC) mice per time point. Data are from one of two experiments that gave similar results. Statistical significance values refer to the total number of DCs in LN, p-values were determined using one-way ANOVA with the Bonferroni post-test. (C) As in B, except that mice were immunized with AF488-Nb instead of Nb. CD86 expression in AF488+ or FITC+ DCs was determined on total MHCIIhi DC populations at the indicated time points. Each dot corresponds to one mouse. Data from AF488-Nb immunization show combination of two separate experiments with three to four mice/group, and data from DBP-FITC treatment are representative of two repeat experiments using six to seven mice/group. P-values were determined using one-way ANOVA with the Bonferroni post-test. (D) C57BL/6 mice were immunized on both ears as indicated. Epidermal layers were harvested for RNA preparation at the indicated times after treatment. Tslp data are expressed as relative to 18S and the respective control samples. Each dot represents one mouse (pool of two ears); mean and SEM for six mice/group are shown. Data are from one of two experiments that gave similar results. P-values were determined using the Kruskal-Wallis with the Dunn’s post-test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.

Characterization of DC subset contribution and Tslp induction during Th2 immune responses to Nb and DBP-FITC. LNs were collected from mice immunized with 600 Nb or DBP-FITC and respective controls at the indicated time points. (A) C57BL/6, Langerin-DTR, and Batf3−/− mice were immunized as indicated. DT was injected i.p. 1 d before and 1 d after immunization. On day 7 after immunization, LN cells were collected and processed for intracellular cytokine staining; in vitro restimulation was with PMA and ionomycin (Nb) or anti-CD3 and anti-CD28 (DBP-FITC). Bar graphs show mean + SEM for 4–8 mice/group. Data are from one of two experiments that gave similar results. P-values were determined using one-way ANOVA with the Bonferroni post-test. (B) C57BL/6 mice were immunized as indicated, and dLN were collected for flow cytometric analysis at the indicated times after treatment. DC subsets were defined as in Fig. S1. Bar graphs show the mean + SE of four (Nb) or seven (DBP-FITC) mice per time point. Data are from one of two experiments that gave similar results. Statistical significance values refer to the total number of DCs in LN, p-values were determined using one-way ANOVA with the Bonferroni post-test. (C) As in B, except that mice were immunized with AF488-Nb instead of Nb. CD86 expression in AF488+ or FITC+ DCs was determined on total MHCIIhi DC populations at the indicated time points. Each dot corresponds to one mouse. Data from AF488-Nb immunization show combination of two separate experiments with three to four mice/group, and data from DBP-FITC treatment are representative of two repeat experiments using six to seven mice/group. P-values were determined using one-way ANOVA with the Bonferroni post-test. (D) C57BL/6 mice were immunized on both ears as indicated. Epidermal layers were harvested for RNA preparation at the indicated times after treatment. Tslp data are expressed as relative to 18S and the respective control samples. Each dot represents one mouse (pool of two ears); mean and SEM for six mice/group are shown. Data are from one of two experiments that gave similar results. P-values were determined using the Kruskal-Wallis with the Dunn’s post-test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.

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