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. 2020 Aug 28;11:1868. doi: 10.3389/fimmu.2020.01868

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Copyright © 2020 Aehnlich, Carnaz Simões, Skadborg, Holmen Olofsson and thor Straten.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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IL-2/IL-15-expanded Vγ9Vδ2 T cells show enhanced cytotoxicity against various cancer cell lines in a 72-h xCELLigence real-time cell analysis assay. (A) Representative cytolysis plot, comparing Vγ9Vδ2 T cell ability to kill FM-28/ZOL between the two expansion conditions. The experiment was conducted at three different effector–target cell ratios (3:1, 1:1, 1:3). Black symbols belong to IL-2-expanded cultures, whereas red symbols indicate IL-2/IL-15-expanded cultures. Lines represent means of three technical replicates. (B) Time to kill 50% of the target tumor cells (KT50) was compared between both expansion conditions at an effector–target cell ratio of 1:1. Horizontal lines depict the median KT50 of the respective expansion conditions (n = 6, compiled from two independent experiments, γδHD7–12). (C) Comparison of percentage of cytolysis at time points 2, 4, and 24 h after addition of IL-2-expanded vs. IL-2/IL-15-expanded Vγ9Vδ2 T cells. Percentage of cytolysis is shown for different effector–target cell ratios. Horizontal lines indicate median cytolysis (n = 6, compiled from two independent experiments, γδHD7–12). (D) Cytolysis plots comparing Vγ9Vδ2 T cell ability to kill FM-55M1/ZOL (left plot), K562 (middle plot), and PC-3/ZOL (right plot) between expansion conditions. Cytotoxicity was assessed at an effector–target cell ratio of 1:1 for FM-55M1/ZOL and PC-3/ZOL and at a ratio of 3:1 for K562. Black, filled symbols indicate IL-2-expanded cultures, whereas matching red, empty symbols indicate the associated IL-2/IL-15-expanded cultures (n = 3 with n = 2 per cell line, compiled from two independent experiments; γδHD9,10,12). % cytolysis and KT50 were calculated using the xCELLigence RTCA Software Pro (ACEA Biosciences). Statistical significance was determined with the Wilcoxon matched-pairs signed-rank test. *p < 0.05. FM-28, FM-55M1 = melanoma cell lines. K562 = chronic myelogenous leukemia cell line. PC-3 = prostate cancer cell line. /ZOL = 24-h-pre-stimulation with ZOL.