Abstract
The anti-tumor efficacy of adoptively transferred T cells requires their in vivo persistence and memory polarization. It is unknown if human chimeric antigen receptor (CAR)-expressing T cells can also undergo memory polarization. We examined the functional status of CAR CD8+ T cells, re-directed to Lewis Y antigen (LeY-T), throughout a period of ex vivo expansion. Immediately before culture CD8+ T cells comprised a mixture of phenotypes including naive (CD45RA+/CCR7+/CD27+/CD28+/perforin−), central memory (CM, CD45RA−/CCR7lo/CD27+/CD28+/perforinlo), effector memory (EM, CD45RA−/CCR7−/CD27+/CD28+/perforinmod) and effector (Eff, CD45RA+/CCR7−/CD27−/CD28−/perforinhi) cells. After transduction and expansion culture of peripheral blood mononuclear cells from normal donors or multiple myeloma patients, CD8+ LeY-T cells polarized to EM- and CM-like phenotype. CD8+ LeY-T cells differed from starting CD8+ CM and EM T cells in that CD27, but not CD28, was downregulated. In addition, CD8+ LeY-T cells expressed high levels of perforin, similar to starting CD8+ Eff. CD8+ LeY-T cells also showed hallmarks of both memory and Eff function, underwent homeostatic proliferation in response to interleukin (IL)-15, and showed interferon (IFN)-γ production and cytotoxicity in response to Le-Y antigen on OVCAR-3 (human ovarian adenocarcinoma) cells. This study confirms CD8+ LeY-T cells have a CM- and EM-like phenotype and heterogeneous function consistent with potential to persist in vivo after adoptive transfer.
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Acknowledgements
Funding for this study came from the National Health and Medical Research Council of Australia, Program Grant no. 454569, the Peter MacCallum Cancer Center Morris Family Grant and the Leukemia Lymphoma Society (6073-06). MH Kershaw and PK Darcy were supported by a NHMRC Senior Research Fellowship and Career Development Award respectively. MJ Smyth and JA Trapani were supported by NHMRC Senior Principal Research Fellowships.
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Neeson, P., Shin, A., Tainton, K. et al. Ex vivo culture of chimeric antigen receptor T cells generates functional CD8+ T cells with effector and central memory-like phenotype. Gene Ther 17, 1105–1116 (2010). https://doi.org/10.1038/gt.2010.59
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DOI: https://doi.org/10.1038/gt.2010.59
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