GM-CSF plus IL-3-derived BMMφ differentiate in a more IL-4 rich environment and produce higher IL-12p40 and IL-4 levels upon stimulation compared to M-CSF-derived BMMφ.

<p>GM-CSF plus IL-3 or M-CSF-derived BMMφ (10⁵/well) from WT (a–f) and IL-4Rα−/− (a, e, and f) mice were primed overnight with IFN-γ and stimulated with LPS (10 ng/ml) in the presence or absence of opsonized SRBC (IC) for 24 hours. (<b>a</b>) IL-4 in the supernatants of the derivation cultures after harvesting of the mature macrophages was assessed by ELISA. p<0.05 by repeated measures ANOVA. (<b>b–d</b>) IL-4, IL-12, and IL-10 levels were assessed in the culture supernatants of GM-CSF plus IL-3 or M-CSF –derived WT macrophages by ELISA. *p<0.01 compared to all other groups (b) or GM-CSF/IL-3 compared to M-CSF (c and d) by one way ANOVA with Bonferroni’s multiple comparison test. (<b>e and f</b>) M-CSF-derived macrophages from WT and IL-4Rα-deficient mice produce similar levels of IL-12 and IL-10 as assessed by ELISA. (<b>a–f</b>) Shown are the means and SEM of 3 independent experiments.