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. 2017 Apr 26;8:502. doi: 10.3389/fimmu.2017.00502

Figure 3.

Figure 3

ISA-2011B impairs CD28 costimulatory functions in Jurkat T cells. (A) NF-AT luciferase activity of Jurkat cells treated for 6 h with DMSO or the indicated concentration of ISA-2011B and stimulated in the absence (med) or presence of 5-3.1/B7 (B7) cells pre-pulsed or not with 1 µg/ml SEB. The results are expressed as the mean of luciferase units ± SD after normalization to GFP values. The data are representative of three independent experiments. (B,D) Real-time PCR was used to measure IL-2 mRNA (B) and IL-8 mRNA levels (D) in Jurkat cells treated with DMSO or the indicated concentration of ISA-2011B and stimulated for 6 h with control IgG or anti-CD28 Abs (D) or anti-CD3 plus anti-CD28 Abs (B). Data are expressed as fold inductions (F.I.) over the basal level of cells stimulated with control IgG and treated with DMSO. Bars show the mean ± SD of three independent experiments. (C) NF-κB luciferase activity of Jurkat cells treated for 6 h with DMSO or the indicated concentration of ISA-2011B and stimulated in the absence (med) or presence of B7-negative (%-3.1) or B7-positive (5-3.1/B7) cells. The results are expressed as the mean of luciferase units ± SD after normalization to GFP values. The data are representative of three independent experiments. Asterisks (*) and (**) indicate P < 0.05 and P < 0.01, respectively, calculated by Student’s t-test.