Generation of CD19-chimeric antigen receptor modified CD8+ T cells derived from virus-specific central memory T cells.
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(+) 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 Ag 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-HSCT, and for comparative studies of CAR and TCR signaling.
Program in Immunology, Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024, USA.
Seitaro
S
Terakura
Tori N
TN
Yamamoto
Rebecca A
RA
Gardner
Cameron J
CJ
Turtle
Michael C
MC
Jensen
Stanley R
SR
Riddell
D019264:Adoptive Transfer; D018941:Antigens, CD19 / Q000235:genetics / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D049109:Cell Proliferation; D005434:Flow Cytometry; D006801:Humans; D007156:Immunologic Memory / Q000276:immunology*; D016219:Immunotherapy, Adoptive; D016086:Lentivirus / Q000235:genetics / Q000276:immunology*; D007938:Leukemia / Q000276:immunology / Q000628:therapy*; D017493:Leukocyte Common Antigens / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D011946:Receptors, Antigen / Q000235:genetics / Q000276:immunology*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology*; D014407:Tumor Cells, Cultured
D018941:Antigens, CD19; D011946:Receptors, Antigen; D011948:Receptors, Antigen, T-Cell; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human