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, After 5 hours, intracellular IFN-? produced by V?2 + T cells was measured by flow cytometry. Values for the percentage of IFN-? producing T cells were compared to control conditions values, p.50
, ?g/ml). (B) iDC were treated by LPS (10 ?g/ml), washed and incubated with V?9V?2 T cells (ratio 1/1) in the presence of neutralizing anti-IL-12 (10 ?g/ml) and anti-IL-18 mAbs
, A polyclonal V?9V?2 T cell line was obtained from PBMC isolated from control and CD212 -(IL-12R?1 -) donors, following a specific amplification (BrHPP 3 ?M, IL-2 at 300 UI/ml), ?g/ml). After 5 hours, intracellular IFN-? produced by V?2 + T cells was measured by flow cytometry. (C)
, /1) which have been sensitized for 2 hours by either LPS (10 ?g/ml) or poly (I:C, p.50
, After 5 hours, intracellular IFN-? produced by V?2 + T cells was measured by flow
, Supplemental Materials and Methods
After 5 hours, cytokine accumulation in ?? T cells, which have been seeded in lower chamber, was determined by intracellular staining. Recombinant human IL-12, IL-18, IL-15, IL-21, IL-23 were obtained from R&D Systems. Blocking antibodies for: IL-12p40/p70 (clones #24910 or #C8.6), IL-18 (clone #125-2H), IFN-? were obtained from BD, R&D Systems and MBL, Mevastatin was purchased from Sigma-Alrich and resupended in DMSO. V?9V?2 Jurkat (JRT3) transfectants were obtained from Dr Julie Déchanet-Merville (CNRS UMR 5164 ,