Abstract
We have succeeded in immortalizing human lymphocytes derived from the peripheral blood of a healthy donor and of an atopic patient, and from the lymph node of a cancer patient by oncogene transfection (Alam et al., 1996). All immortalized human lymphocytes were shown to be CD3+ and CD19−, indicating that these immortalized human lymphocytes were all T cells. We established 317, 154 and 692 individual immortalized human T cell lines derived from the healthy donor, the atopic patient and the cancer patient, respectively. The ratios of CD4+ and CD8+ subpopulations within the set containing immortalized T cells derived from the healthy donor were shown to be varied depending on the combinations of transfected oncogenes used. However, CD8+ cells were found to be the dominant subpopulation of immortalized T cells derived from the atopic patient and the cancer patient. These immortalized T cells showed different proliferative responses in the presence of exogenous human IL–2 depending on their origin, and was consistent with the surface expression of the IL–2 receptor. Furthermore, the cytokine secretion patterns of these immortalized T cells stimulated with mitogen were investigated. The results showed that the immortalized T cells from the healthy donor is able to secrete various kinds of cytokines such as IL–2, IL–10, β-IFN and GM-CSF. However, immortalized T cells from the cancer patient was shown to only secrete IL–2 and GM-CSF. These results suggest that depending on the origin, the immortalized T cells came from different subsets or from cells in different activated states. Mixed lymphocytes reactions demonstrated that these immortalized T cells are able to proliferate in the presence of allogenic or xenogenic stimulator cells, suggesting that they maintain the ability to recognize specific antigens on the stimulator cells and can proliferate even after the immortalization. Furthermore, immortalized T cells derived from the healthy donor and the cancer patient strongly responded to K562 cells, suggesting that MHC-nonrestricted killer T cells were also immortalized.
Abbreviations IL–2R – interleukin 2 receptor; MLR – mixed lymphocyte reaction
Similar content being viewed by others
References
Alam S, Katakura Y, Yoshida H, Shirahata S and Murakami H (1996) In vitro immortalization of human T lymphocytes by oncogenes. Animal Cell Technology: Basic and Applied Aspects Vol. 8 (ed. by Funatsu K et al.) (in press), Kluwer Academic Press, Dordrecht, The Netherlands.
Biesinger B, Müller-Fleckenstein I, Simmer B, Lang G, Wittmann S, Platzer E, Desrosiers RC and Fleckenstein B (1992) Stable growth transformation of human T lymphocytes by Herpesvirus saimiri. Proc Natl Acad Sci USA 89: 3116–3119.
Borek C (1980) X-ray-induced in vitro neoplasmic transformation of human diploid cells. Nature 283: 776–778.
De Carli M, Berthold S, Fickenscher H, Fleckenstein IM, D'Elios MM, Gao Q, Biagiotti R, Giudizi MG, Kalden JR, Fleckenstein B, Romagnani S and Del Prete G (1993) Immortalization with Herpesvirus saimiri modulates the cytokine secretion profile of established Th1 and Th2 human T cell clones. J Immunol 151: 5022–5030.
Karasuyama H, Tohyama N and Toda T (1989) Autocrine growth and tumorigenicity of interleukin 2-dependent helper T cells transfected with IL-2 gene. J Exp Med 169: 13–25.
Murakami H, Okazaki Y, Yamada K and Omura H (1988) Egg yolk lipoprotein, a new supplement for the growth of mammalian cells in serum-free medium. Cytotechnology 1: 159–169.
Okano F, Tachibana H, Akiyama K, Shirahata S and Murakami H (1993) Immortalization of human T lymphocytes by oncogenes. Cytotechnology 11: 205–211.
Pisa P, Halapi E, Pisa EK, Gerdin E, Hising C, Bucht A, Gerdin B and Kiessling R (1992) Selective expression of interleukin 10, interferon γ, and granulocyte-macrophage colony-stimulating factor in ovarian cancer biopsies. Proc Natl Acad Sci USA 89: 7708–7712.
Russo J, Reina D, Frederick J and Russo IH (1988) Expression of phenotypical changes by human breast epithelial cells treated with carcinogens in vivo. Cancer Res 48: 2837–2857.
Schwartz RH (1992) Costimulation of T lymphocytes: The role of CD28, CTLA-4, and B7/BB1 in interleukin 2 production and immunotherapy. Cell 71: 1065–1068.
Shay JW and Wright WE (1989) Quantitation of the frequency of immortalization of normal human diploid fibroblasts by SV40 large T-antigen. Exp Cell Res 184: 109–118.
Stevenson M, Volsky B, Hedenskog M and Volsky DJ (1986) Immortalization of human T lymphocytes after transfection of Epstein-Barr virus DNA. Science 233: 981–983.
Troidl B, Simmer B, Fickenscher H, Muller-Fleckenstein I, Emmrich F, Fleckenstein B and Gebhart E (1994) Karyotypic characterization of human T cell lines immortalized by Herpes virus saimiri. Int J Cancer 56: 433–438.
van Neerven RJJ, van 't Hof W, Ringrose JH, Jansen HM, Aalberse RC, Wierenga EA and Kapsenberg ML (1993) T cell epitopes of house dust mite major allergen Der pII. J Immunol 151: 2326–2335.
Yano T, Teruya K, Shirahata S, Watanabe J, Osada K, Tachibana H, Ohashi H, Kim E-H and Murakami H (1994) Ras oncogene enhances the production of a recombinant protein regulated by the cytomegalovirus promoter in BHK-21 cells. Cytotechnology 16: 167–178.
Zupo S, Azzoni L, Massara R, D'Amato A, Perussia B and Ferrarini M (1993) Coexpression of Fcγ receptor IIIA and interleukin 2 receptor β chain by a subset of human CD3+/CD8+/CD11b+ lymphocytes. J Clin Immunol 13: 228–236.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Alam, S., Katakura, Y., Yoshida, H. et al. Functional characterization of human T cells immortalized by oncogene transfection. Cytotechnology 23, 185–192 (1997). https://doi.org/10.1023/A:1007928021139
Issue Date:
DOI: https://doi.org/10.1023/A:1007928021139