IL-23 Regulates Virulence Figure 5. IL-23 producing DCs induced by opportunistic fungi drive Th17 responses. (A) IL-17 and IFN-c production determined by ELISA in memory T cell cultures primed for 5 d in plates coated with anti-CD3 and anti-CD28, in the presence of supernatants (sups) of unstimulated DCs, or sups of DCs stimulated with either Aspergillus spores, or hyphae (above plots), and then re-stimulated for 24 h with anti-CD3 and anti-CD28. Data are expressed as mean 6 SEM values for DCs derived from six different donors. (B) Intracellular cytokine staining for IL-17 and IFN-c in memory CD4+ T cells primed and expanded as described in (A), and re-stimulated for 5 h with PMA and ionomycin. (C) ELISA of IL-17 and IFN-c in 24-hour culture supernatants (sups) of naive and memory CD4+ T cells primed for 5 d in plates coated with anti-CD3 and anti-CD28, in the presence of sups of unstimulated DCs or Aspergillus spores-stimulated DCs or Aspergillus-hyphae stimulated DCs alone or in the presence of neutralizing anti-IL-23 antibodies. Data shown are representative of 5 independent experiments. Error bars represent SD. *, P,0.001, paired Student’s t test. doi:10.1371/journal.pone.0012955.g005 PLoS ONE | www.plosone.org 7 September 2010 | Volume 5 | Issue 9 | e12955