Vγ9Vδ2 Tcells are non-canonical Tcells that use their Tcell receptor to detect phosphoantigens bound to the internal domain of the HMBPP receptor (butyrophilin 3/2A1 complex). This protocol describes the expansion and purification of human effector Vγ9Vδ2 Tcells from human buffy coat and describes how to assess their activation by antigen-containing target cells. While specifically focused on cytokine production, this protocol can be readily adapted to evaluate other effector functions of activated Vγ9Vδ2 Tcells. For complete details on the use and execution of this protocol, please refer to Hsiao etal. (2022) and Hsiao and Wiemer (2018).

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