Figure 5.

IL-18 and N-acetylcysteine enable Vγ9Vδ2 T cell proliferation under severe lymphopenic conditions. (A) Enriched γδ T cells devoid of CD14⁺ monocytes were obtained from peripheral blood mononuclear cells (PBMCs) using CD14 microbeads and magnetic depletion using LD columns. (B) Photo documentation of cell cultures: enriched γδ T cells were seeded at 10³ cells per 96 well and stimulated with 10 μM geranylgeranyl pyrophosphate (GGPP) plus 100 U/mL IL-2 and 100 ng/mL IL-18 in the presence or absence of N-acetylcysteine (NAC). Scale bar: 500 μm. (C) Enriched γδ T cells were seeded in triplicates at 10³ cells per 96 well and stimulated with 10 μM mevalonate-derived isoprenoid pyrophosphates plus 100 U/mL IL-2 and NAC in the absence or presence of 100 ng/mL IL-18 . After 14 days, cells were stained with fluorophore-conjugated antibodies CD3 and Vδ2 to confirm γδ T-cell identity. Absolute cell numbers and γδ T-cell expansion were determined by cytofluorimetric analysis (FACS). Data are representative of 4 independent experiments with T cells derived from 4 different donors.