CD4⁺ as well as CD8⁺ αβT cells are reprogrammed with a γ9δ2TCR. (A) Mock and γ9δ2TCR-transduced αβT cells were stained with γδTCR, CD4- and CD8-specific Abs, and analyzed by flow cytometry. Percentage of positive cells for each quadrant is indicated. (B) CD4⁺ and CD8⁺ αβT cells were isolated and incubated with Daudi cells with 10μM pamidronate for 48 hours. IFNγ secretion was measured by ELISPOT. The test was performed in duplicate for every condition in 3 independent experiments. (C) CD4⁺ (95% purity), CD8⁺ (90% purity), or mixed–total γ9δ2TCR-transduced αβT cells or mock cells were incubated with ⁵¹Cr-loaded Daudi or PBMCs at the indicated E:T ratios for 4 hours and the specific lysis was calculated. Shown are representative experiments of at least 3 independent repetitions. (D) Immature DCs were incubated with mock CD4⁺ or γ9δ2TCR-transduced CD4⁺ αβT cells without or with pamidronate (10μM) for 48 hours at ratio of 1:2 (iDC: αβT cells). Shown is a representative FACS analysis of CD80, CD86, HLA-DR, CD83, and CD40.