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. Author manuscript; available in PMC: 2011 Dec 1.
Published in final edited form as: Scand J Immunol. 2010 Dec;72(6):511–521. doi: 10.1111/j.1365-3083.2010.02467.x

Figure 1. Antigen-activated but not naïve T cells induce IL-12p75 from DCs.

Figure 1

(A) BM-DCs at 2×104 cells/well from B10.A-RAG2KO mice were cocultured for 48 hr with 1×105 sorted naïve or antigen-activated 5C.C7 T cells +0.1μM MCC peptide in a 96-well U-bottom plate. LPS was added at 200ng/ml to DCs either at time zero (T=0) or 24h (T=−24h) and washed before the addition of T cells. ELISA (R&D system) specifically measured IL-12p75 in CSN after 48h. These data are expressed as the mean ± SD triplicates from a 96-well plate. (B–E) Multiplex cytokine array (SearchLight) measuring various cytokines in CSN obtained from LPS-activated (T=−24h) B10.A-RAG2KO. BMDCs at 2×104 cells/well cocultured for 48 hr with 1×105 naïve or antigen-activated 5C.C7 T cells +0.1μM MCC peptide in a 96 well U-bottom plate. These data are expressed as the mean ± SD composite of three independent experiments. (F) Same as (A) except two populations of DCs were used: 1-LPS-preactivated DCs (T=−24h) were either cocultured with naïve T cells plus 100ng/ml of rIFN-γ or with antigen-activated T cells plus 5μg/ml of neutralizing antibody for IFN-γ or 5μg/ml an isotype control Gt-anti-mouse. 2-“Resting” DC were stimulated with LPS or LPS plus IFN-γ stimulation.