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. 2013 Apr 9;4:1685. doi: 10.1038/ncomms2684

Figure 1. PGE2-cAMP signalling induces IL-12Rβ2 expression in TCR-activated T cells.

Figure 1

(a) Expression of Ifng, Tbx21 and Il12rb2 mRNA by T cells activated for indicated times with antibody to CD3 and antibody to CD28 (αCD3/CD28) in the absence or presence of PGE2 under Th1-priming conditions. A portion of cells were restimulated with PMA and ionomycin for the last 4 h (72R). (b) Expression of Ifng, Tbx21 and Il12rb2 mRNA by T cells activated for 24 h with αCD3/CD28 in the absence or presence of PGE2 or selective agonists to EP1 to EP4 under Th1-priming conditions. (c) Surface expression of IL-12Rβ2 in T cells activated for 48 h with αCD3/CD28 in the absence or presence of PGE2 or agonists selective to EP1 to EP4 under Th1-priming conditions. Grey-filled histogram represents isotype control. ΔMFI (mean fluorescence intensity) indicates the differences between MFI of IL-12Rβ2 and MFI of isotype control (right). (d) Time-course of Il12rb2 mRNA expression by PGE2 in T cells activated with αCD3/CD28. (e) PGE2 induces IL-12Rβ2 protein expression in T cells activated with αCD3/CD28 for 48 h. (f,g) Expression of Il12rb2 mRNA in WT T cells (f) or IFN-γR1–/– T cells supplemented with anti-IL-2 (g), activated for 24 h with αCD3/CD28 in the absence or presence of PGE2 with or without Wortmannin, LY-294002 or H-89. (h) Expression of Il12rb2 mRNA in T cells activated for 24 h with αCD3/CD28 in the absence or presence of PGE2 with H-89, Rp-8-Br-CAMPS or Rp-8-CPT-CAMPS. (i,j) Expression of IL-12Rβ2 mRNA (i) and protein (j) in T cells activated with αCD3/CD28 in the presence of db-cAMP, forskolin or 3-isobutyl-1- methylxanthine (IBMX) for 12 h (i) or 48 h (j). (k) Expression of Il12rb2 mRNA in IFN-γR1−/− CD4+ T cells activated for 24 h with αCD3/CD28 with or without db-cAMP in the absence or presence of anti-IL-2 or LY-294002 (LY) or both. Data shown as mean±s.e.m. are representative of two or more independent experiments with triplicates. Statistical significance was examined by unpaired two-tailed Student’s t-test, *P<0.05; **P<0.01; ***P<0.001. NS, not significant, veh, vehicle; a.u., arbitrary units.