Production of Human T-Cell Hybridomas by Electrofusion

  • C. Gravekamp
  • S. J. L. Bol
  • A. Hagemeijer
  • R. L. H. Bolhuis


The somatic cell hybridization technique provides a means for the immortalization of specific cellular functions. Köhler and Milstein (1975) were the first to be successful in the immortalization of the production of specific immunoglobulins by cell fusion. They prepared a continuously immunoglobulin-secreting hybridoma cell line by fusion of mouse myeloma cells with spleen cells of an immunized mouse in the presence of inactivated Sendai virus. In addition to Sendai virus, polyethylene glycol (PEG) has also been widely used for cell fusion. During the past 10 years, an explosive development of hybridoma technology has taken place in many laboratories for the in vitro production of mouse monoclonal antibodies [for a review, see Reading (1982)]. It was later demonstrated that other cellular functions can also be immortalized by cell fusion. For example, mouse T-cell hybridomas that secrete immunoregulatory molecules such as interleukin 2 (IL-2) or T-cell replacing factor (TRF) have been produced. Other mouse T-cell hybrids expressing helper or suppressor functions have been established [for a review, see Malissen and Zeurthen (1982)]. More recently, several laboratories have directed their attention to xenogeneic and human—human hybrids exerting specific immunologic functions with a view to their potential clinical application. However, the preparation of human hybrids has generally appeared to be more difficult than that of murine hybrids. Initial growth of the fused cells (for human T-T and for some human B—B cell hybridomas) is difficult to achieve and often it takes more than a month before proliferation of fused cells can be observed (Greene et al., 1982; Grillot-Courvalin and Brouet, 1981; Sikora et al., 1982; Chiorazzi et al.,1982). Moreover, human—human hybridomas frequently show chromosomal instability (Foung et al., 1982; Kozbor and Roder, 1983). In this respect, the choice of fusion agent could be an important factor. PEG has a number of disadvantages that might be drawbacks in the production of a larger repertoire of hybrid cells.


Cell Fusion Hybrid Cell Fusion Product Cytolytic Activity Parental Cell Line 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Abo, T., and Balch, T., 1981, A differentiation antigen of human NK and K cells identified by a monoclonal antibody (HNK-1), J. Immunol. 127: 1024–1029.PubMedGoogle Scholar
  2. Ahkong, Q. F., Fisher, D., Tampion, W., and Lucy, A. J., 1975, Mechanisms of cell fusion, Nature 253: 194–196.PubMedCrossRefGoogle Scholar
  3. Berebbi, M., Foa, C., Imbert, E., Fabre, I., and Lidcey, C., 1983, Cytolytically active murine T lymphocyte/polyoma virus transformed fibroblast hybrids, Exp. Cell Res. 145: 357–368.PubMedCrossRefGoogle Scholar
  4. Berke, G., 1983, Cytotoxic T lymphocytes, how do they function? Immunol. Rev. 72: 5–42.PubMedCrossRefGoogle Scholar
  5. Burwen, S. J., and Satir, B. H., 1977, A freeze-fracture study of early membrane events during mast cell secretion, J. Cell. Biol. 73: 660–671.PubMedCrossRefGoogle Scholar
  6. Chiorazzi, N., Wasserman, R. J., and Kunkel, H. G., 1982, Use of Epstein—Barr-virus transformed B cell lines for the generation of immunoglobulin-producing human B cell hybridoma, J. Exp. Med. 156: 930–935.PubMedCrossRefGoogle Scholar
  7. Foung, S. H. K., Sasaki, D. T., Grumet, F. C., and Engleman, E. G., 1982, Production of functional human T-T cell hybridomas in selection medium lacking aminopterin and thymidine, Proc. Natl. Acad. Sci. USA 79: 7484–7488.PubMedCrossRefGoogle Scholar
  8. Greene, W. C., Fleisher, A., Nelson, D. L., and Waldmann, T. A., 1982, Production of human T suppressor hybridomas, J. Immunol. 129 (5): 1986–1992.PubMedGoogle Scholar
  9. Grillot-Courvalin, C., and Brouet, J. C., 1981, Establishment of a human T cell hybrid line with suppressive activity, Nature 292 (27): 844–845.PubMedCrossRefGoogle Scholar
  10. Hayata, M., Oshimura, J., Minowada, J., and Sandberg, A. A., 1975, Chromosomal banding of cultured T and B lymphocytes, In Vitro 11 (6): 361–368.PubMedCrossRefGoogle Scholar
  11. Kaufmann, Y., Berke, G., and Eshhar, Z., 1981, Functional cytotoxic T lymphocyte hybridomas, Transplant. Proc. XIII (1): 1170–1174.Google Scholar
  12. Köhler, G., and Milstein, C., 1975, Continuous cultures of fused cells secreting antibodies of predefined specificity, Nature 256: 495.PubMedCrossRefGoogle Scholar
  13. Kozbor, D., and Roder, J. C., 1983, The production of monoclonal antibodies from human lymphocytes, Immunol. Today 4 (3): 72–79.CrossRefGoogle Scholar
  14. Malissen, B., and Zeurthen, J., 1982, Immortalizing T-cell function, Immunol. Today 3 (4): 94–95.CrossRefGoogle Scholar
  15. Mukherji, B., and Cieplinski, W., 1983, Functional hybrids between human cytotoxic T and mouse myeloma cells, Hybridoma 2 (4): 383–392.PubMedCrossRefGoogle Scholar
  16. Nabholz, M., Cianfriglia, M., Acuto, O., Conzelmann, A., Haas, W., Boehmer, H. V., McDonald, H. R., Pohlit, H., and Johnson, J. P., 1980, Cytolytically active murine T cell hybrids, Nature 287: 437–439.PubMedCrossRefGoogle Scholar
  17. Oi, V. T., and Herzenberg, L. A., 1980, Immunoglobulin-producing hybrid cell lines, in: Selected Methods in Cellular Immunology ( B. B. Mishell and S. M., Shigii, eds.), Freeman, San Francisco, pp. 351–372.Google Scholar
  18. Reading, C. L., 1982, Theory and methods for immunization in culture and monoclonal antibody production, J. Immunol. Meth. 53: 261–291.CrossRefGoogle Scholar
  19. Roos, S. D., Robinson, J. M., and Davidson, R. L., 1983, Cell fusion and intramembrane particle distribution in polyethelene glycol resistant cells, J. Cell Biol. 97: 909–917.PubMedCrossRefGoogle Scholar
  20. Sikora, K., Alderson, T., Phillips, J., and Watson, J. V., 1982, Human hybridomas from malignant gliomas, Lancet 1: 11–14.PubMedCrossRefGoogle Scholar
  21. Uchiyama, T., Broder, S., and Waldman, T. A., 1981, A monoclonal antibody (anti-TAC) reactive with activated and functionally mature human T cells, J. Immunol. 126: 1393.PubMedGoogle Scholar
  22. Van de Griend, R.J., and Bolhuis, R. L. H., 1984, Rapid expansion of allospecific cytotoxic T cell clones using nonspecific feeder cell lines without further addition of exogenous IL-2, Transplantation, 38 (4): 401–406.PubMedCrossRefGoogle Scholar
  23. Vienken, J., Zimmermann, U., Fouchard, M., and Zagury, D., 1983, Electrofusion of myeloma cells on the single level, FEBS Lett. 163 (1): 54–56.PubMedCrossRefGoogle Scholar
  24. Zimmermann, U., 1982, Electric field mediated fusion and related electrical phenomena, Biochim. Biophys. Acta 694: 227–277.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • C. Gravekamp
    • 1
  • S. J. L. Bol
    • 1
  • A. Hagemeijer
    • 2
  • R. L. H. Bolhuis
    • 1
  1. 1.Rotterdam Radiotherapeutic InstituteRotterdamThe Netherlands
  2. 2.Department of Cell Biology and GeneticsErasmus UniversityRotterdamThe Netherlands

Personalised recommendations