Transfer of a TCR gene derived from a patient with a marked antitumor response conveys highly active T-cell effector functions

Marybeth S. Hughes, Yik Y.L. Yu, Mark E. Dudley, Zhili Zheng, Paul F. Robbins, Yong Li, John Wunderlich, Robert G. Hawley, Morvarid Moayeri, Steven A. Rosenberg, Richard A. Morgan

Research output: Contribution to journalArticle

Abstract

The genes for the α and β chains of a highly reactive anti-MART-1 T-cell receptor were isolated from T-lymphocytes that mediated in vivo regression of tumor in a patient with metastatic melanoma. These genes were cloned and inserted into MSCV-based retroviral vectors. After transduction, greater than 50% gene transfer efficiency was demonstrated in primary T-lymphocytes stimulated by an anti-CD3 antibody. The specificity and biologic activity of TCR gene-transduced T-cells was determined by cytokine production after coculture of T-cells with stimulator cells pulsed with MART-1 peptide. The production of interferon-γ and granulocyte macrophage-colony stimulating factor (GM-CSF) was comparable to highly active MART-1 specific peripheral blood lymphocytes (PBL) in the amount of cytokine produced and transduced cells recognized peptide pulsed cells at dilutions similar to cytotoxic T lymphocyte (CTL) clones. Human leukocyte antigen (HLA) class I restricted recognition was demonstrated by mobilization of degranulation marker CD107a, by cell lysis, by cytokine production, and by proliferation in the presence of HLA-A2-positive but not HLA-A2-negative melanoma cell lines. Similar data was obtained when tumor-infiltrating lymphocytes (TIL) were transduced with the TCR genes, converting previously nonreactive cells to tumor reactive cells. TCR-transduced T-cells are thus attractive candidates for evaluation in cell transfer therapies of patients with cancer.

Original languageEnglish (US)
Pages (from-to)457-472
Number of pages16
JournalHuman gene therapy
Volume16
Issue number4
DOIs
StatePublished - Apr 1 2005

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ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Hughes, M. S., Yu, Y. Y. L., Dudley, M. E., Zheng, Z., Robbins, P. F., Li, Y., Wunderlich, J., Hawley, R. G., Moayeri, M., Rosenberg, S. A., & Morgan, R. A. (2005). Transfer of a TCR gene derived from a patient with a marked antitumor response conveys highly active T-cell effector functions. Human gene therapy, 16(4), 457-472. https://doi.org/10.1089/hum.2005.16.457