Figure 2

A PFKFB3 small molecule antagonist inhibits stimulation of F2,6BP, glucose uptake, lactate secretion, ATP and TNF-α secretion caused by 10 hours of exposure to anti-CD3/anti-CD28-conjugated microbeads. (A), Either vehicle (DMSO) or 1–10 μM 3PO was added to human CD3+ T cells stimulated with anti-CD3/anti-CD28-conjugated microbeads for 5, 10 and 24 hours and F2,6BP was determined. (B), 2-[1-¹⁴ C]-deoxy-D-glucose uptake after addition of anti-CD3/anti-CD28-conjugated microbeads in the presence or absence of the indicated concentrations of 3PO for 5, 10 and 24 hours. (C), Lactate secretion after addition of anti-CD3/anti-CD28-conjugated microbeads in the presence or absence of the indicated concentrations of 3PO for 5, 10 and 24 hours. (D), Intracellular ATP after addition of anti-CD3/anti-CD28-conjugated microbeads in the presence or absence of the indicated concentrations of 3PO for 5, 10 and 24 hours. (E), Either vehicle (DMSO) or 1–10 μM 3PO was added to human CD3⁺ T cells stimulated with anti-CD3/anti-CD28-conjugated microbeads for 10 hours and cell aggregates were assessed using light microscopy. (F), Secreted TNF-α after addition of anti-CD3/anti-CD28-conjugated microbeads in the presence or absence of the indicated concentrations of 3PO for 5, 10 and 24 hours was determined by ELISA. Data are representative of three independent experiments. *p < 0.05 (relative to vehicle control).