@article{2fc44e50416648ad9cb4cc3a347b08e2,
title = "Generation and application of TGFβ-educated human Vγ9Vδ2 T cells",
abstract = "Clinical trials that tested the antitumor activity of γδ T cells have been mostly unsuccessful. To address this, we expanded human Vγ9Vδ2 T cells in TGFβ1, a cytokine which enhances their viability, trafficking properties, and intrinsic antitumor activity. This protocol summarizes the production and in vitro functional characterization of TGFβ1 educated human Vγ9Vδ2 cells and highlights their compatibility with chimeric antigen receptor (CAR) engineering. We also describe in vivo testing of the antitumor activity of these CAR T cells in mice. For complete details on the use and execution of this protocol, please refer to Beatson et al. (2021).",
keywords = "Biotechnology and bioengineering, Cancer, Cell Biology, Cell culture, Cell-based Assays, Flow Cytometry/Mass Cytometry, Health Sciences, Immunology, Model Organisms, Molecular Biology",
author = "Parente-Pereira, {Ana C.} and Beatson, {Richard E.} and Davies, {David M.} and Caroline Hull and Whilding, {Lynsey M.} and Porter, {Joanna C.} and John Maher",
note = "Funding Information: This study was funded by Bloodwise (Programme 13007 ), Cancer Research UK (CRUK)/ Bayer ( C11499 / A19444 ), the Medical Research Council ( DPFS MR / M024733/1 and MR/R000026/1 ), a Medical Research Council Confidence in Concepts award, a CRUK Multidisciplinary Project Award ( C48390/A21153 ), the British Lung Foundation ( MESOUK18-2 ), the Experimental Cancer Medicine Centre at King{\textquoteright}s College London , the King{\textquoteright}s Health Partners/ King{\textquoteright}s College London Cancer Research UK Cancer Centre, Leucid Bio and by the National Institute for Health Research (NIHR) Biomedical Research Centre based at Guy{\textquoteright}s and St Thomas{\textquoteright} NHS Foundation Trust , and King{\textquoteright}s College London (grant number IS-BRC-1215-20006 ). The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health. We thank many colleagues for generously providing cell lines. Funding Information: This study was funded by Bloodwise (Programme 13007), Cancer Research UK (CRUK)/ Bayer (C11499/A19444), the Medical Research Council (DPFS MR/ M024733/1 and MR/R000026/1), a Medical Research Council Confidence in Concepts award, a CRUK Multidisciplinary Project Award (C48390/A21153), the British Lung Foundation (MESOUK18-2), the Experimental Cancer Medicine Centre at King's College London, the King's Health Partners/ King's College London Cancer Research UK Cancer Centre, Leucid Bio and by the National Institute for Health Research (NIHR) Biomedical Research Centre based at Guy's and St Thomas{\textquoteright} NHS Foundation Trust, and King's College London (grant number IS-BRC-1215-20006). The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health. We thank many colleagues for generously providing cell lines. A.P-P. R.B. D.D. C.H. and L.W. conducted experimental work. A.P-P. R.B. D.D. C.H. L.W. and J.M. designed and optimized the approach and interpreted the data. R.B. and J.M. wrote the first draft of the manuscript. All authors reviewed and edited the manuscript. J.M. is scientific founder, chief scientific officer, and shareholder in Leucid Bio which is a spin-out company focused on development of cellular therapeutic agents. J.M. A.P-P. and R.B. are named inventors on granted patents pertaining to this work. L.W. and D.M.D. were both formerly consultants to Leucid Bio. J.M. and L.W. are named inventors on a granted patent submitted in relation to this work while D.D. C.H. L.W. and J.M. are named inventors on submitted patents that relate to pCAR technology. D.M.D. is a shareholder in Leucid Bio. J.C.P. declares no competing financial interests. Publisher Copyright: {\textcopyright} 2022 The Author(s)",
year = "2022",
month = jun,
day = "17",
doi = "10.1016/j.xpro.2022.101319",
language = "English",
volume = "3",
pages = "101319",
journal = "STAR Protocols",
publisher = "Cell Press",
number = "2",
}
