Sleeping Beauty transposon-mediated engineering of human primary T cells for therapy of CD19+ lymphoid malignancies.
We have reported earlier that the non-viral Sleeping Beauty (SB) transposon system can mediate genomic integration and long-term reporter gene expression in human primary peripheral blood (PB) T cells. In order to test whether this system can be used for genetically modifying both PB T cells and umbilical cord blood (UCB) T cells as graft-versus-leukemia effector cells, an SB transposon was constructed to coexpress a single-chain chimeric antigen receptor (CAR) for human CD19 and CD20. PB and UCB were nucleofected with the transposon and a transposase plasmid, activated and then expanded in culture using anti-CD3/CD28 beads. Stable dual-gene expression was confirmed in both T-cell types, permitting enrichment by positive selection with Rituxan. The engineered CD4(+) T cells and CD8(+) T cells both exhibited specific cytotoxicity against CD19(+) leukemia and lymphoma cell lines, as well as against CD19 transfectants, and produced high-levels of antigen-dependent Th1 (but not Th2) cytokines. The in vivo adoptive transfer of genetically engineered T cells significantly reduced tumor growth and prolonged the survival of the animal. Taken together, these data indicate that T cells from PB and UCB can be stably modified using a non-viral DNA transfer system, and that such modified T cells may be useful in the treatment of refractory leukemia and lymphoma.
Huang
Xin
X
Division of Blood and Marrow Transplantation, Department of Pediatrics, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
Guo
Hongfeng
H
Kang
Johnthomas
J
Choi
Suet
S
Zhou
Tom C
TC
Tammana
Syam
S
Lees
Christopher J
CJ
Li
Zhong-Ze
ZZ
Milone
Michael
M
Levine
Bruce L
BL
Tolar
Jakub
J
June
Carl H
CH
Scott McIvor
R
R
Wagner
John E
JE
Blazar
Bruce R
BR
Zhou
Xianzheng
X
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D058846:Antibodies, Monoclonal, Murine-Derived; D018941:Antigens, CD19 / Q000235:genetics*; D018951:Antigens, CD20 / Q000235:genetics; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006801:Humans; D016219:Immunotherapy, Adoptive; D007938:Leukemia / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D008223:Lymphoma / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D008957:Models, Genetic; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D018626:Retroelements / Q000235:genetics*; D000069283:Rituximab; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D014162:Transfection / Q000379:methods
D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D018941:Antigens, CD19; D018951:Antigens, CD20; D018626:Retroelements; D000069283:Rituximab