Generation of CD19-chimeric antigen receptor modified CD8+ T cells derived from virus-specific central memory T cells

S Terakura, TN Yamamoto, RA Gardner… - Blood, The Journal …, 2012 - ashpublications.org
S Terakura, TN Yamamoto, RA Gardner, CJ Turtle, MC Jensen, SR Riddell
Blood, The Journal of the American Society of Hematology, 2012ashpublications.org
The adoptive transfer of donor T cells that have been genetically modified to recognize
leukemia could prevent or treat leukemia relapse after allogeneic HSCT (allo-HSCT).
However, adoptive therapy after allo-HSCT should be performed with T cells that have a
defined endogenous TCR specificity to avoid GVHD. Ideally, T cells selected for genetic
modification would also have the capacity to persist in vivo to ensure leukemia eradication.
Here, we provide a strategy for deriving virus-specific T cells from CD45RA− CD62L+ CD8+ …
Abstract
The adoptive transfer of donor T cells that have been genetically modified to recognize leukemia could prevent or treat leukemia relapse after allogeneic HSCT (allo-HSCT). However, adoptive therapy after allo-HSCT should be performed with T cells that have a defined endogenous TCR specificity to avoid GVHD. Ideally, T cells selected for genetic modification would also have the capacity to persist in vivo to ensure leukemia eradication. Here, we provide a strategy for deriving virus-specific T cells from CD45RACD62L+CD8+ central memory T (TCM) cells purified from donor blood with clinical grade reagents, and redirect their specificity to the B-cell lineage marker CD19 through lentiviral transfer of a gene encoding a CD19-chimeric Ag receptor (CAR). Virus-specific TCM were selectively transduced by exposure to the CD19 CAR lentivirus after peptide stimulation, and bi-specific cells were subsequently enriched to high purity using MHC streptamers. Activation of bi-specific T cells through the CAR or the virus-specific TCR elicited phosphorylation of downstream signaling molecules with similar kinetics, and induced comparable cytokine secretion, proliferation, and lytic activity. These studies identify a strategy for tumor-specific therapy with CAR-modified T cells after allo-HSCT, and for comparative studies of CAR and TCR signaling.
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