Sleeping beauty transposon-mediated engineering of human primary T cells for therapy of CD19+ lymphoid malignancies

Xin Huang, Hongfeng Guo, Johnthomas Kang, Suet Choi, Tom C. Zhou, Syam Tammana, Christopher J. Lees, Zhong Ze Li, Michael Milone, Bruce L. Levine, Jakub Tolar, Carl H. June, R. Scott McIvor, John E. Wagner, Bruce R. Blazar, Xianzheng Zhou

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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.

Original languageEnglish (US)
Pages (from-to)580-589
Number of pages10
JournalMolecular Therapy
Volume16
Issue number3
DOIs
StatePublished - Jan 1 2008

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Beauty
Human Engineering
Cell- and Tissue-Based Therapy
T-Lymphocytes
Neoplasms
Fetal Blood
Blood Cells
Leukemia
Lymphoma
Transposases
Gene Expression
Antigen Receptors
Adoptive Transfer
Reporter Genes
Plasmids
Cytokines
Transplants
Antigens
Cell Line

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Sleeping beauty transposon-mediated engineering of human primary T cells for therapy of CD19+ lymphoid malignancies. / Huang, Xin; Guo, Hongfeng; Kang, Johnthomas; Choi, Suet; Zhou, Tom C.; Tammana, Syam; Lees, Christopher J.; Li, Zhong Ze; Milone, Michael; Levine, Bruce L.; Tolar, Jakub; June, Carl H.; McIvor, R. Scott; Wagner, John E.; Blazar, Bruce R.; Zhou, Xianzheng.

In: Molecular Therapy, Vol. 16, No. 3, 01.01.2008, p. 580-589.

Research output: Contribution to journalArticle

Huang, Xin ; Guo, Hongfeng ; Kang, Johnthomas ; Choi, Suet ; Zhou, Tom C. ; Tammana, Syam ; Lees, Christopher J. ; Li, Zhong Ze ; Milone, Michael ; Levine, Bruce L. ; Tolar, Jakub ; June, Carl H. ; McIvor, R. Scott ; Wagner, John E. ; Blazar, Bruce R. ; Zhou, Xianzheng. / Sleeping beauty transposon-mediated engineering of human primary T cells for therapy of CD19+ lymphoid malignancies. In: Molecular Therapy. 2008 ; Vol. 16, No. 3. pp. 580-589.
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