Cytokine-independent growth and clonal expansion of a primary human CD8+ T-cell clone following retroviral transduction with the IL-15 gene

Cary Hsu, Stephanie A. Jones, Cyrille J. Cohen, Zhili Zheng, Keith Kerstann, Juhua Zhou, Paul F. Robbins, Peter D. Peng, Xinglei Shen, Theotonius J. Gomes, Cynthia E. Dunbar, David J. Munroe, Claudia Stewart, Kenneth Cornetta, Danny Wangsa, Thomas Ried, Steven A. Rosenberg, Richard A. Morgan

Research output: Contribution to journalArticle

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Abstract

Malignancies arising from retrovirally transduced hematopoietic stem cells have been reported in animal models and human gene therapy trials. Whether mature lymphocytes are susceptible to insertional mutagenesis is unknown. We have characterized a primary human CD8+ Tcell clone, which exhibited logarithmic ex vivo growth in the absence of exogenous cytokine support for more than 1 year after transduction with a murine leukemia virus-based vector encoding the T-cell growth factor IL-15. Phenotypically, the clone was CD28 -, CD45RA-, CD45RO+, and CD62L-, a profile consistent with effector memory T lymphocytes. After gene transfer with tumor-antigen-specific T-cell receptors, the clone secreted IFN-γ upon encountering tumor targets, providing further evidence that they derived from mature lymphocytes. Gene-expression analyses revealed no evidence of insertional activation of genes flanking the retroviral insertion sites. The clone exhibited constitutive telomerase activity, and the presence of autocrine loop was suggested by impaired cell proliferation following knockdown of IL-15Rα expression. The generation of this cell line suggests that nonphysiologic expression of IL-15 can result in the long-term in vitro growth of mature human T lymphocytes. The cytokine-independent growth of this line was a rare event that has not been observed in other IL-15 vector transduction experiments or with any other integrating vector system. It does not appear that the retroviral vector integration sites played a role in the continuous growth of this cell clone, but this remains under investigation.

Original languageEnglish
Pages (from-to)5168-5177
Number of pages10
JournalBlood
Volume109
Issue number12
DOIs
StatePublished - Jun 15 2007

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Interleukin-15
T-cells
Clone Cells
Genes
Cytokines
T-Lymphocytes
Lymphocytes
Growth
Insertional Mutagenesis
Clone cells
Gene transfer
Gene therapy
Mutagenesis
Telomerase
Cell proliferation
Neoplasm Antigens
T-Cell Antigen Receptor
Murine Leukemia Viruses
Stem cells
Viruses

ASJC Scopus subject areas

  • Hematology

Cite this

Cytokine-independent growth and clonal expansion of a primary human CD8+ T-cell clone following retroviral transduction with the IL-15 gene. / Hsu, Cary; Jones, Stephanie A.; Cohen, Cyrille J.; Zheng, Zhili; Kerstann, Keith; Zhou, Juhua; Robbins, Paul F.; Peng, Peter D.; Shen, Xinglei; Gomes, Theotonius J.; Dunbar, Cynthia E.; Munroe, David J.; Stewart, Claudia; Cornetta, Kenneth; Wangsa, Danny; Ried, Thomas; Rosenberg, Steven A.; Morgan, Richard A.

In: Blood, Vol. 109, No. 12, 15.06.2007, p. 5168-5177.

Research output: Contribution to journalArticle

Hsu, C, Jones, SA, Cohen, CJ, Zheng, Z, Kerstann, K, Zhou, J, Robbins, PF, Peng, PD, Shen, X, Gomes, TJ, Dunbar, CE, Munroe, DJ, Stewart, C, Cornetta, K, Wangsa, D, Ried, T, Rosenberg, SA & Morgan, RA 2007, 'Cytokine-independent growth and clonal expansion of a primary human CD8+ T-cell clone following retroviral transduction with the IL-15 gene', Blood, vol. 109, no. 12, pp. 5168-5177. https://doi.org/10.1182/blood-2006-06-029173
Hsu, Cary ; Jones, Stephanie A. ; Cohen, Cyrille J. ; Zheng, Zhili ; Kerstann, Keith ; Zhou, Juhua ; Robbins, Paul F. ; Peng, Peter D. ; Shen, Xinglei ; Gomes, Theotonius J. ; Dunbar, Cynthia E. ; Munroe, David J. ; Stewart, Claudia ; Cornetta, Kenneth ; Wangsa, Danny ; Ried, Thomas ; Rosenberg, Steven A. ; Morgan, Richard A. / Cytokine-independent growth and clonal expansion of a primary human CD8+ T-cell clone following retroviral transduction with the IL-15 gene. In: Blood. 2007 ; Vol. 109, No. 12. pp. 5168-5177.
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AU - Hsu, Cary

AU - Jones, Stephanie A.

AU - Cohen, Cyrille J.

AU - Zheng, Zhili

AU - Kerstann, Keith

AU - Zhou, Juhua

AU - Robbins, Paul F.

AU - Peng, Peter D.

AU - Shen, Xinglei

AU - Gomes, Theotonius J.

AU - Dunbar, Cynthia E.

AU - Munroe, David J.

AU - Stewart, Claudia

AU - Cornetta, Kenneth

AU - Wangsa, Danny

AU - Ried, Thomas

AU - Rosenberg, Steven A.

AU - Morgan, Richard A.

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N2 - Malignancies arising from retrovirally transduced hematopoietic stem cells have been reported in animal models and human gene therapy trials. Whether mature lymphocytes are susceptible to insertional mutagenesis is unknown. We have characterized a primary human CD8+ Tcell clone, which exhibited logarithmic ex vivo growth in the absence of exogenous cytokine support for more than 1 year after transduction with a murine leukemia virus-based vector encoding the T-cell growth factor IL-15. Phenotypically, the clone was CD28 -, CD45RA-, CD45RO+, and CD62L-, a profile consistent with effector memory T lymphocytes. After gene transfer with tumor-antigen-specific T-cell receptors, the clone secreted IFN-γ upon encountering tumor targets, providing further evidence that they derived from mature lymphocytes. Gene-expression analyses revealed no evidence of insertional activation of genes flanking the retroviral insertion sites. The clone exhibited constitutive telomerase activity, and the presence of autocrine loop was suggested by impaired cell proliferation following knockdown of IL-15Rα expression. The generation of this cell line suggests that nonphysiologic expression of IL-15 can result in the long-term in vitro growth of mature human T lymphocytes. The cytokine-independent growth of this line was a rare event that has not been observed in other IL-15 vector transduction experiments or with any other integrating vector system. It does not appear that the retroviral vector integration sites played a role in the continuous growth of this cell clone, but this remains under investigation.

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