Payload-delivering engineered γδ T cells display enhanced cytotoxicity, persistence, and efficacy in preclinical models of osteosarcoma

T cell-based cancer immunotherapy has typically relied on membrane-bound cytotoxicity enhancers such as chimeric antigen receptors expressed in autologous αβ T cells. These approaches are limited by tonic signaling of synthetic constructs and costs associated with manufacturing. γδ T cells are an emerging alternative for cellular therapy, possessing innate anti-tumor activity, potent antibody-dependent cellular cytotoxicity, and minimal alloreactivity. We present an immunotherapeutic platform technology built around the innate properties of the Vγ9Vδ2 T cell, harnessing specific characteristics of this cell type and offering an allo-compatible cellular therapy that recruits bystander immunity. We engineered γδ T cells to secrete synthetic tumor-targeting opsonins in the form of an scFv-Fc fusion protein and a mitogenic IL-15Ra–IL-15 fusion protein (stIL15). Using GD2 as a model antigen, we show that GD2-specific opsonin-secreting Vγ9Vδ2 T cells (stIL15-OPS-γδ T cells) have enhanced cyto..., , , # γδ T cells engineered to secrete immune-active payloads display enhanced persistence and efficacy in preclinical models of osteosarcoma

[https://doi.org/10.5061/dryad.q2bvq83t1](https://doi.org/10.5061/dryad.q2bvq83t1)

2 datasets are provided,  one pertaining to the main figures (Dataset 1) and one to the supplementary. figures (Dataset 2).  The datasets comprise tabulated data for all of the figures shown.

## Description of the data and file structure

Data is shown in tabulated Excel files, with each tab pertaining to a panel in the figures

**Data File S1**

1B: Dead cell % for SUP-T1-wt targets or SUP-T1-GD2 targets co-cultured with Unmodified gdT cells or OPS-gdT cells, as determined by flow cytometry

1C: Absolute numbers of gdT cells ± engineering at different days of expansion in the conditions indicated in the columns

1D-E: Differences in expression or indicated markers between unmodified-gdT cells or 14G2a stIL15-OPS-gdT cells expressed as Earth Movers Distance values, d...