Role of F1-ATPase and NKG2D in the target recognition by γ9δ2TCR-transduced αβT cells. (A) Typical FACS histograms of Daudi cells and PBMCs stained with specific Abs for F1-ATPase α and β subunit followed with secondary goat–anti-mouse–PE. Secondary Ab alone was used as a negative control staining. (B-C) Daudi cells were incubated with blocking Abs either for the α subunit or β subunit of F1-ATPase (B) or for NKG2D (C) and γ9δ2TCR-transduced αβT cells for 48 hours with or without pamidronate (100μM). IFNγ secretion was measured by ELISPOT. The experiment was performed in duplicate and statistical analyses were performed using the Student t test. Similar results were obtained in at least 3 independent experiments; *P < .01; **P < .001. (D-E) γ9δ2TCR-transduced NKG2D⁻ and NKG2D⁺ γ9δ2TCR- or mock-transduced αβT cells were incubated with Daudi in the absence (□) or presence of 10μM pamidronate () for 48 hours. Then IFNγ and TNFα secretion was measured by ELISPOT. Shown are representative results of 3 independent experiments.