Frequencies of polyfunctional alloreactive T cells are increased after stimulation with FL B cells with dual TNFRSF14 aberrations. (A) Proinflammatory cytokines generated from allogeneic T cells after primary (n = 9) and secondary (n = 7) coculture with TNFRSF14 WT or dual TNFRSF14 aberration FL B cells. (B) Intracellular cytokine accumulation in allogeneic CD4⁺ and CD8⁺ T cells after primary (n = 8) and secondary (n = 7) coculture with FL B cells. Cytokine-positive cell frequencies are expressed as a percentage of T-cell subsets. (C) Surface CD107a⁺ cell frequencies (expressed as a percentage of T-cell subsets) in allogeneic CD4⁺ and CD8⁺ T cells after primary (n = 8) and secondary (n = 7) coculture with FL B cells. (A-C) Mean values ± standard deviation are shown. (D) Frequencies of allogeneic bifunctional T cells (CD107a⁺ and cytokine⁺) after primary allogeneic coculture with FL B cells. Horizontal lines depict median values. (E) Multifunctional allogeneic T-cell effectors after primary coculture with FL B cells. Cytokine-secreting populations within the alloreactive CD107a⁺ T-cell compartment are shown. Arcs show frequencies of T cells positive for CD107a, IFN-γ, TNF-α, and IL-2, and slices show effector populations with combined expression patterns of CD107a, IFN-γ, TNF-α, and IL-2. Mean frequencies from 7 independent experiments are shown. P values are for 2-tailed Student t tests throughout.