Ca²⁺ supplementation <i>in vitro</i> modulates cytokine production by expanded human <i>i</i>NKT cells and Vδ2⁺ T cells.

(a, b)iNKT cells were expanded ex vivo in the presence of αGalCer. The expanded cells were stimulated for 4 h with 25 ng/ml PMA and 1 μg/ml ionomycin in either normal RPMI1640 medium (RPMInorm) or RPMI1640 medium supplemented with 1 mM Ca2+ (RPMIsuppl). Vα24i NKT cells (live CD14- CD20- CD3+ 6B11+ cells) were analysed for the expression of (a) CD69 and the production of (b) TNF. (c-e) Vδ2+ T cells were expanded in vitro in the presence of Zoledronic acid. Human Vδ2+ T cells (live CD14- CD20- CD3+ Vδ2+ cells) were analysed for the production of (c) IL-2, (d) IFNγ, and (e) GM-CSF. Summary graphs (left panels) and representative data (right panels) from gated iNKT cells and Vδ2+ T cells are shown, respectively. Data were pooled from four (iNKT cells; n = 12) and three (Vδ2+ T cells, n = 9) independent experiments with three samples each.