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

Publication Type:

Journal Article


Blood, Volume 119, Issue 1, p.72-82 (2012)


2012, Adoptive Transfer, ANTIGEN, Antigens, CD19, Antigens, CD45, CD8-Positive T-Lymphocytes, Cell Proliferation, Center-Authored Paper, Clinical Research Division, Cultured, Cytokine Analysis Core Facility, Flow Cytometry Core Facility, Humans, Immunologic Memory, Immunotherapy, Adoptive, Lentivirus, LEUKEMIA, Lymphocyte Activation, Nov 11, November 2011, Receptors, Antigen, T-Cell, Tumor Cells


The adoptive transfer of donor T cells that have been genetically modified to recognize leukemia could prevent or treat leukemia relapse after allogeneic hematopoietic stem cell transplant (allo-HCT). However, adoptive therapy after allo-HCT should be performed with T cells that have a defined endogenous T cell receptor (TCR) specificity to avoid graft-versus-host disease. 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(+) central memory T (T(CM)) 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 antigen receptor (CAR). Virus-specific T(CM) 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-HCT, and for comparative studies of CAR and TCR signaling.