Abstract
Peripheral blood T cells transduced with a tumor-specific T-cell receptor (TCR) face problems of auto-reactivity and lack of efficacy caused by cross-pairing of exogenous and endogenous TCR chains, as well as short term in vivo survival due to activation and growth factor-induced differentiation. We here studied an alternative strategy for the efficient generation of naive CD8+ T cells with a single TCR. TCR-transduced human postnatal thymus-derived and adult mobilized blood-derived hematopoietic progenitor cells (HPCs) were differentiated to CD4+CD8+ double-positive T cells using OP9-Delta-like 1 (OP9-DL1) cultures. Addition of the agonist peptide induced double positive cells to cross-present the peptide, leading, in the absence of co-stimulation, to cell cycle arrest and differentiation into mature CD8+ T cells. Comprehensive phenotypic, molecular and functional analysis revealed the generation of naive and resting CD8+ T cells through a process similar to thymic positive selection. These mature T cells show a near complete inhibition of endogenous TCRA and TCRB rearrangements and express high levels of the introduced multimer-reactive TCR. Upon activation, specific cytokine production and efficient killing of tumor cells were induced. Using this strategy, large numbers of high-avidity tumor-specific naive T cells can be generated from readily available HPCs without TCR chain cross-pairing.
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Acknowledgements
The authors would like to thank the staff of the Cardiac Surgery and Hematology department and the Cell Processing Lab of Ghent University Hospital for providing thymus and MB samples. We are indebted to Katleen De Preter and Frank Speleman of the Center for Medical Genetics (Ghent University) for providing mRNA expression data of thymic subsets. Finally, we would like to thank Dr T Boterberg for the irradiation of the feeder cells, Karin Weening for help with the retroviral constructs, Sophie Vermaut for help with flow cytometry and cell sorting and Christian De Boever for the art work.
Disclosures
This work was supported by the Research Foundation - Flanders (Fonds voor Wetenschappelijk Onderzoek Vlaanderen, FWO), Stichting tegen Kanker, the geconcerteerde onderzoeksactiviteiten of Ghent University, and the Interuniversity Attraction Poles Program supported by the Belgian Science Policy. YVC, SV and GV are supported by the Instituut voor de Aanmoediging van Innovatie door Wetenschap en Technologie in Vlaanderen. SS, TT and TK are supported by the Research Foundation-Flanders (FWO).
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Snauwaert, S., Verstichel, G., Bonte, S. et al. In vitro generation of mature, naive antigen-specific CD8+ T cells with a single T-cell receptor by agonist selection. Leukemia 28, 830–841 (2014). https://doi.org/10.1038/leu.2013.285
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DOI: https://doi.org/10.1038/leu.2013.285
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