Abstract
Although Epstein-Barr virus (EBV) infection elicits both CD4- and CD8-positive T-cell responses, a considerable body of experimental evidence indicates that the latter responses play a major role in controlling the number of EBV-infected cells and proliferation of EBV-transformed B-blasts in vivo (1,2). Recently, cytotoxic T-cells (CTLs) were introduced into clinical practice as an efficient tool for treatment of EBV-associated malignancies (3,4). This chapter is focused on describing the generation and characterization of EBV-specific CD8-positive cytotoxic T-cells. Primarily, techniques that are used to generate CTLs specific for EBV proteins expressed in EBV-transformed lymphoblastoid cell lines (LCLs) will be discussed.
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References
Masucci, M. G. and Ernberg, I. (1994) Epstein-Barr virus: adaptation to a life within the immune system. Trends Microbiol. 2, 125–130.
Rickinson, A. B. and Kieff, E. (1996) Epstein-Barr virus, in Fields Virology, vol. 2 (Fields, B. N., Knipe, D. M., and Howley, P. M., eds.), Lippincott-Raven, Philadelphia, pp. 2397–2446.
Heslop, H. E., Ng, C. Y. C., Li, C., Smith, C. A., Loftin, S. K., Krance, R. A., et al. (1996) Long-term restoration of immunity against Epstein-Barr virus infection by adoptive transfer of gene-modified virus-specific T lymphocytes. Nature Med. 2, 551–555.
Rooney, C. M., Smith, C. A., Ng, C. Y. C., Loftin, S., Li, C., Krance, R. A., Brenner, M. K., and Heslop, H. E. (1995) Use of gene-modified virus-specific T lymphocytes to control Epstein-Barr-virus-related lymphoproliferation. Lancet 345, 9–13.
Coligan, T. E., KruisBeck, A. M., Margulies, D. H., and Strobek, W. (1996) Current Protocols in Immunology, in 3 volumes, National Institutes of Health, John Wiley and Sons Inc., New York, NY.
Fries, K. L., Sculley, T. B., Webster-Cyriaque, J., Rajadurai, P., Sadler, R. H., and Raab-Traub, N. (1997) Identification of a novel protein encoded by the BamHI A region of the Epstein-Barr virus. J. Virol. 71, 2765–2771.
Rabin, H., Hopkins, R. F., Ruscetti, F. W., Neubawer, R. H., Brown, R. L., and Kawakami, T. G. (1981) Spontaneous release of a factor with properties of T-cell growth factor from a continuous line of primate tumor T cells. J. Immunol. 127, 1852–1856.
Levitsky, V., Zhang, Q.-J., Levitskaja, J., and Masucci, M. G. (1996) The life span of major histocompatibility complex-peptide complexes influences the efficiency of presentation and immunogenicity of two class I-restricted cytotoxic T lymphocyte epitopes in the Epstein-Barr virus nuclear antigen 4. J. Exp. Med. 183, 915–926.
Khanna, R., Burrows, S. R., Neisig, A., Neefjes, J., Moss, D. J., and Silins, S. L. (1997) Hierarchy of Epstein-Barr virus-specific cytotoxic T-cell responses in individuals carrying different subtypes of an HLA allele: implications for epitope-based antiviral vaccines. J. Virol. 71, 7429–7435.
Hill, A. B., Lee, S. P., Haurum, J. S., Murray, N., Yao, Q.-Y., Rowe, M., et al. (1995) Class I major histocompatibility complex-restricted cytotoxic T lymphocytes specific for Epstein-Barr virus (EBV) nuclear antigens fail to lyse the EBV-transformed B lymphoblastoid cell lines against which they were raised. J. Exp. Med. 181, 2221–2228.
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© 2001 Humana Press Inc., Totowa, NJ
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Levitsky, V., Frisan, T., Masucci, M. (2001). Generation of Polyclonal EBV-Specific CTL Cultures and Clones. In: Wilson, J.B., May, G.H.W. (eds) Epstein-Barr Virus Protocols. Methods in Molecular Biology™, vol 174. Humana Press. https://doi.org/10.1385/1-59259-227-9:203
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DOI: https://doi.org/10.1385/1-59259-227-9:203
Publisher Name: Humana Press
Print ISBN: 978-0-89603-690-1
Online ISBN: 978-1-59259-227-2
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