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. 2015 Jul 6;10(7):e0132348. doi: 10.1371/journal.pone.0132348

Fig 7. BMDCs incubated in vitro with the A2BR agonist produce increased amounts of IL-23 and IL-6 and have enhanced ability enhancing Th17 response.

Fig 7

A) BMDCs cultured from immunized B6 mice were further cultured for 48h in the absence or presence of A2BR agonist (100 ng/ml), then IL-1, IL-12, IL-23 and IL-6 levels in the culture supernatants were measured by ELISA. B) Neither TLR2 ligand nor A2BR agonist along induced increased MHC II and CD86 expression on BMDCs, but a synergistic effect did. Cultured BMDCs were incubated with or without TLR2 ligand (Pam3CSK, 10μg/ml) and in the presence or absence of the A2BR agonist (100 ng/ml). 24h later, the expression of MHC II and CD86 on BMDCs was assessed by FACS after staining with FITC-anti-mouse MHC II or FITC-anti-mouse CD86 antibodies, respectively. C) The 48h supernatants of T cell-BMDC co-cultures described in Fig 7B were tested for IL-17 amount. **p<0.01. D) After an in vitro treatment with vehicle (control) or A2BR agonist for 24 h, the supernatants of BMDCs were taken and added to co-cultures of in vivo primed CD3+ responder T cells and splenic APCs, in the presence of the immunizing peptide and under Th17 polarizing conditions. After 5 days, proliferating T cells were stained intracellularly for IL-17.