title: Sleeping Beauty transposon-mediated engineering of human primary T cells for therapy of CD19+ lymphoid malignancies.
abstract: 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.
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Pediatrics, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Huang
authors.forename: Hongfeng
authors.initials: H
authors.lastname: Guo
authors.forename: Johnthomas
authors.initials: J
authors.lastname: Kang
authors.forename: Suet
authors.initials: S
authors.lastname: Choi
authors.forename: Tom C
authors.initials: TC
authors.lastname: Zhou
authors.forename: Syam
authors.initials: S
authors.lastname: Tammana
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Lees
authors.forename: Zhong-Ze
authors.initials: ZZ
authors.lastname: Li
authors.forename: Michael
authors.initials: M
authors.lastname: Milone
authors.forename: Bruce L
authors.initials: BL
authors.lastname: Levine
authors.forename: Jakub
authors.initials: J
authors.lastname: Tolar
authors.forename: Carl H
authors.initials: CH
authors.lastname: June
authors.forename: R
authors.initials: R
authors.lastname: Scott McIvor
authors.forename: John E
authors.initials: JE
authors.lastname: Wagner
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Blazar
authors.forename: Xianzheng
authors.initials: X
authors.lastname: Zhou
mesh_terms: 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
chemical_list: D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D018941:Antigens, CD19; D018951:Antigens, CD20; D018626:Retroelements; D000069283:Rituximab