American Association for Cancer Research
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FIGURE 2 from Exercise-induced β2-adrenergic Receptor Activation Enhances the Antileukemic Activity of Expanded γδ T-Cells via DNAM-1 Upregulation and PVR/Nectin-2 Recognition

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posted on 2024-05-13, 14:20 authored by Forrest L. Baker, Kyle A. Smith, Preetesh L. Mylabathula, Tiffany M. Zúñiga, Douglass M. Diak, Helena Batatinha, Grace M. Niemiro, Michael D. Seckeler, Charles R. Pedlar, Daniel P. O'Connor, Jamie Colombo, Emmanuel Katsanis, Richard J. Simpson

Acute exercise enhances the antileukemic activity of ex vivo expanded Vγ9Vδ2+ T-cells in vitro and in vivo. A, Schematic of the experimental design for in vitro and in vivo experiments. B, The total number (1 × 106) of Vγ9Vδ2+ cells isolated before (REST) and during the final 5 minutes of exercise (EX) at day 0, and the total number of Vγ9Vδ2+ T-cells generated in the expanded cell products after stimulation with ZOL+IL2 for 14 days. The number of Vγ9Vδ2+ cells generated at day 14 relative to the number of γδ cells in the PBMC fractions at day 0. The cellular composition of the expanded Vγ9Vδ2+ T-cell products at day 14. The proportions of naïve (N), CM, EM, and CD45RA+ EMRA cells among Vγ9Vδ2+ T-cells after 14 days of expansion (n = 10). C, The in vitro anti-tumor activity of ex vivo expanded Vγ9Vδ2+ T-cells against K562, Daudi, and U266 with and without ZOL sensitization. Target cells were exposed to 5 µmol/L of ZOL for 20 hours and cytotoxicity of Vγ9Vδ2+ cells was assessed via flow cytometry–based assays. D, To determine the in vivo GvL effect of exercise expanded Vγ9Vδ2+, NSG-IL15 mice were injected with REST or EX expanded Vγ9Vδ2+ (10 × 106) and challenged with 1 × 106 luciferase tagged human chronic myeloid leukemia cells (K562-luc). A subset of mice in each group were also provided weekly injections of ZOL (REST+ZOL; EX+ZOL; VEHICLE +ZOL). The BLI (photons/second) scores, TFS, change in BW, and overall probability of survival (OS) after injection of expanded Vγ9Vδ2+ T-cells with or without ZOL sensitization (n = 8–11/group). BLI significant difference from EX+ZOL and EX conditions indicated by * and #, respectively (all conditions were significantly different from vehicle controls). TFS, BW, and OS significant difference from vehicle controls were indicated by *. E, Representative bioluminescence images of leukemia-bearing mice that received (from left to right) vehicle, vehicle + ZOL, REST expanded Vγ9Vδ2+ T-cells, REST expanded Vγ9Vδ2+ T-cells + ZOL, EX expanded Vγ9Vδ2+ T-cells, and EX expanded Vγ9Vδ2+ T-cells + ZOL. BLI intensity on a scale from low (purple) to high (red). Data are represented as mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by Student two-tailed paired t test (B); repeated measures two-way ANOVA or LMM with Bonferroni post hoc test (C–D); log-rank (Mantel–Cox) test (D).


HHS | NIH | National Cancer Institute (NCI)

American College of Sports Medicine (ACSM)

University of Arizona Cancer Center (UACC)

People Acting Now Discover Awards (PANDA)

HHS | NIH | National Heart, Lung, and Blood Institute (NHLBI)



Exercise mobilizes cytotoxic lymphocytes to blood which may allow superior cell products to be harvested and manufactured for cancer therapy. Gamma-Delta (γδ) T-cells have shown promise for treating solid tumors, but there is a need to increase their potency against hematologic malignancies. Here, we show that human γδ T-cells mobilized to blood in response to just 20 minutes of graded exercise have surface phenotypes and transcriptomic profiles associated with cytotoxicity, adhesion, migration, and cytokine signaling. Following 14 days ex vivo expansion with zoledronic acid and IL2, exercise mobilized γδ T-cells had surface phenotypes and transcriptomic profiles associated with enhanced effector functions and demonstrated superior cytotoxic activity against multiple hematologic tumors in vitro and in vivo in leukemia-bearing xenogeneic mice. Infusing humans with the β1+β2-agonist isoproterenol and administering β1 or β1+β2 antagonists prior to exercise revealed these effects to be β2-adrenergic receptor (AR) dependent. Antibody blocking of DNAM-1 on expanded γδ T-cells, as well as the DNAM-1 ligands PVR and Nectin-2 on leukemic targets, abolished the enhanced antileukemic effects of exercise. These findings provide a mechanistic link between exercise, β2-AR activation, and the manufacture of superior γδ T-cell products for adoptive cell therapy against hematologic malignancies. Exercise mobilizes effector γδ T-cells to blood via β2-adrenergic signaling which allows for generation of a potent expanded γδ T-cell product that is highly cytotoxic against hematologic malignancies.