Figure 3.

Phenotype and functional characterisation of expanded γδ-T cells. characterisation of the effector/memory, activation, and exhaustion (A) profile of CD40L/pp65 expanded γδ-T cells in the CD4⁺, CD8⁺, and CD4^neg/CD8^neg subpopulations. (B) show the cytolytic activity by ⁵¹Cr release assay at different E:T ratios of γδ aAPC/pp65 or aAPC/CD40L/pp65 (black line) compared to polyclonally activated αβ-T cells (black dotted line) against Daudi tumour cells and aCD19⁺ B cells. (C) shows long-term cytotoxic activity of γδ CD40L/pp65 T cells against Daudi tumour cells in 3 or 6 days of co-culture at E:T ratios of 1:1 and 5:1 compared to polyclonally activated αβ-T cells. 6 day co-culture with Daudi tumour cells and γδ CD40L/pp65 T cells in presence of cytokines (IL2/IL15). Degranulation assay evaluating the CD107a expression of γδ CD40L/pp65 in co-culture with a panel of different tumour cell lines (D). ELISA assay measuring the IFN-γ release in the supernatant collected from co-culture experiments (E). (F) shows the number of spot forming units per 1 × 10⁶ γδ-T cells in IFN-γ Elispot when co-cultured with aCD19⁺ B cells or EBV infected B cells. Data are summarised as average ± SEM of 4 donors. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.