Summary
Antigen-specific T cells play a key role in cellular immune response against cancer. The ability to isolate, maintain, and characterize tumor-specific T cells is a prerequisite to studying anticancer immune response and developing novel strategies for cancer immunotherapy. However, the life span of human T cells in vitro is usually short and is limited by the onset of cellular senescence. To establish long-term, antigen-specific T-cell lines and clones, we selectively immortalized antigen-responsive T cells from human peripheral blood mononuclear cells (PBMCs). PBMCs were stimulated with antigens, and then infected with a murine leukemia virus-based retroviral vector carrying an immortalizing gene, the human telomerase-reverse transcriptase gene. Since such vectors can only integrate in dividing cells, only antigen-activated T cells are efficiently transduced. Using this approach, we generated immortalized T-cell lines that maintained strictly IL-2-dependent growth and MHC-restricted, antigen-specific responsiveness, some of which have been in continuous culture for longer than 1 year, far in excess of the survival of parallel control nonimmortalized cultures. These lines showed antigen-specific proliferation with induced cytokine and chemokine production, and, in the case of CD8+ T-cell lines, antigen-specific cytolytic activity. When applied to the tumor antigen-specific T cells, the approach provides a convenient, reproducible means for generating a stable, continuously renewable source of antigen-specific T lymphocytes for a variety of studies on anticancer immunity.
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Acknowledgments
The author would like to thank David Ott, Claes Öhlén, and Jeffrey Lifson for their advice and encouragement and Matthew Trivett for excellent technical assistance. This project has been funded in whole or in part with Federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. NO1-CO-12400. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.
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Barsov, E.V. (2009). Selective Immortalization of Tumor-Specific T Cells to Establish Long-Term T-Cell Lines Maintaining Primary Cell Characteristics. In: Kozlov, S.V. (eds) Inflammation and Cancer. Methods in Molecular Biology™, vol 511. Humana Press. https://doi.org/10.1007/978-1-59745-447-6_6
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DOI: https://doi.org/10.1007/978-1-59745-447-6_6
Publisher Name: Humana Press
Print ISBN: 978-1-934115-14-5
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