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 language | English (US) |
|---|---|
| Pages (from-to) | 580-589 |
| Number of pages | 10 |
| Journal | Molecular Therapy |
| Volume | 16 |
| Issue number | 3 |
| DOIs | |
| State | Published - Jan 1 2008 |
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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 journal › Article
}
TY - JOUR
T1 - Sleeping beauty transposon-mediated engineering of human primary T cells for therapy of CD19+ lymphoid malignancies
AU - Huang, Xin
AU - Guo, Hongfeng
AU - Kang, Johnthomas
AU - Choi, Suet
AU - Zhou, Tom C.
AU - Tammana, Syam
AU - Lees, Christopher J.
AU - Li, Zhong Ze
AU - Milone, Michael
AU - Levine, Bruce L.
AU - Tolar, Jakub
AU - June, Carl H.
AU - McIvor, R. Scott
AU - Wagner, John E.
AU - Blazar, Bruce R.
AU - Zhou, Xianzheng
PY - 2008/1/1
Y1 - 2008/1/1
N2 - 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.
AB - 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.
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U2 - 10.1038/sj.mt.6300404
DO - 10.1038/sj.mt.6300404
M3 - Article
VL - 16
SP - 580
EP - 589
JO - Molecular Therapy
JF - Molecular Therapy
SN - 1525-0016
IS - 3
ER -