Figure 4.

ISA-2011B impairs TCR- and CD28-induced increase of [Ca²⁺]_i and IL-2 gene expression in primary T cells. (A,B) Primary T cells from HD were treated for 6 h with DMSO or ISA-2011B (10 µM) and after loading T cells with Fluo-3-AM, Ca²⁺ levels ([Ca²⁺]_i) were measured by cytofluorimetric analysis after stimulation with crosslinked anti-CD3 plus anti-CD28 Abs (A) or A23871 (2 µg/ml) for 10 min. Data are representative of three independent experiments. (C) Real-time PCR was used to measure IL-2 mRNA levels in primary T cells from healthy donors (n = 5) treated with DMSO or ISA-2011B (10 µg/ml) and stimulated for 6 h with crosslinked anti-CD3 plus anti-CD28 Abs. Values, normalized on GAPDH, are expressed as arbitrary units. Median values: DMSO = 0.55; DMSO CD3 plus CD28 = 16.59; ISA-2011B = 0.25: ISA-2011B CD3 plus CD28 = 5.68. Lines represent median values and P values are indicated where significant (Mann–Whitney).