Transfection as an Approach to Understanding Membrane Glycoproteins

  • A. Fortunato
  • R. F. L. James
  • A. Mellor
  • N. A. Mitchison
Conference paper
Part of the Haematology and Blood Transfusion / Hämatologie und Bluttransfusion book series (HAEMATOLOGY, volume 28)


Gene transfection has much to contribute to our understanding of membrane glycoproteins. The technique is in principle simple: it consists of the transfer of a single gene from one cell to another, using the method of DNA recombination in plasmids to manipulate the gene during the transfer and to rescue it for analysis afterwards. This is valuable for several reasons. The first and simplest is that it generates a cell which has a single new gene product. As the functions of most gene products are still unknown, this should greatly help us to find out what these functions are. For example, the function of the great majority of membrane glycoproteins such as Thy 1, Lyt 1, and T5/T8 remains to be understood. Most membrane glycoproteins have so far been defined only as antigens, sometimes and to an increasing extent through the use of monoclonal antibodies. It turns out to be very difficult to find out what these glycoproteins do, even after quite a lot has been found out about their structure. The Thy 1 molecule is a case in point. It was discovered 18 years ago, it has been used as a marker in lymphocyte differentiation for 13 years, and its primary structure has now been unravelled [23], yet we still know next to nothing about its function. Up to a point the classical approaches of genetics can be applied to these problems: analysis by means of loss and temperature-sensitive mutations. Nowadays these may be supplemented by segregation analysis, in which a cell positive for a given glycoprotein and a given function is fused with a negative cell and the daughter cells analysed for co-expression of the glycoprotein and the function [3]. But progress using classical genetics has been slow, and the contribution to be exprected from transfection is accordingly great. With some justice one could argue that transfection is not a new departure in principle since it merely uses positive variants where classical genetics uses negative variants. However there are many reasons for expecting these positive variants to be far more valuable.


Membrane Glycoprotein Congenic Strain Associative Recognition Classical Genetic Congenic 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. 1.
    Bailey DW (1975) Genetics of histocompatibility in mice. I. New loci and congenic lines. Immunogenetics 2:249–256CrossRefGoogle Scholar
  2. 2.
    Boon T, Van Snick J, Van Pel A, Uyttenhove C, Marchand M (1980) Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815. II. T lymphocyte-mediated cytolysis. J Exp Med 152:1184–1193PubMedCrossRefGoogle Scholar
  3. 3.
    Bramwell ME, Harris H (1978) An abnormal membrane glycoprotein associated with malignancy in a wide range of different tumours. Proc R Soc Lond [Biol] 210:87–106CrossRefGoogle Scholar
  4. 4.
    Bromberg J, Brenan M, Clark E, Lake P, Mitchison NA, Nakashima I, Sainis K (1979) Associative recognition in the response to alloantigens (and xenogenisation to alloantigens). Gan 23:185–192Google Scholar
  5. 5.
    Butcher GW, Corvalan JR, Licence DR, Howard JC (1982) Immune response genes controlling responsiveness to major transplantation antigens. Specific major histocompatibility complex-linked defect for antibody response to Class I alloantigens. J Exp Med 155:303–320PubMedCrossRefGoogle Scholar
  6. 6.
    Clark EA, Lake P, Favila-Castillo L (1981) Modulation of Thy-1 alloantibody responses: donor cell-associated H-2 inhibition and augmentation without recipient Ir gene control. J Immunol 127:2135–2140PubMedGoogle Scholar
  7. 7.
    Dresser DW, Popham AM, Hunt R (1982) Differences in putative minor histocompatibility but not Igh genes can prevent T-cell priming and T-B cooperation in the response of mice to sheep erythrocytes. Immunology 46:643–651PubMedGoogle Scholar
  8. 8.
    Engelhard YH, Kaufman JF, Strominger JL, Burakoff SJ (1980) Specificity of mouse cytotoxic T lymphocytes stimulated with either HLA-A and -B or HLA-DR antigens reconstituted into phospholipid vesicles. J Exp Med 152:54s–64sPubMedCrossRefGoogle Scholar
  9. 9.
    Klein J, Juretic A, Baxevanis CN, Nagy ZA (1981) The traditional and a new version of the mouse H-2 complex. Nature 291:455–460PubMedCrossRefGoogle Scholar
  10. 10.
    Kripke M (1974) Antigenicity of murine skin tumours induced by ultraviolet light. J Natl Cancer Inst 53: 1333–1336PubMedGoogle Scholar
  11. 11.
    Lake P, Douglas TC (1978) Recognition and genetic control of helper determinant for cell surface antigen Thy-1. Nature 275:220–222PubMedCrossRefGoogle Scholar
  12. 12.
    Mellor AL, Golden L, Weiss E, Bullman H, Hurst J, Simpson E, James R, Townsend ARM, Taylor PM, Schmidt W, Ferluga J, Leben L, Santamaria M, Atfield G, Festenstein H, Flavell RA (1982) Expression of the murine H-2Kb histocompatibility antigen in cells transformed with cloned H-2 genes. Nature 298:529–533PubMedCrossRefGoogle Scholar
  13. 13.
    Mitchison NA (1979) Regulation of the response to cell surface antigens. In: Ferrone S, Gorini S, Herberman RB, Reisfeld RA (eds) Current trends in tumor immunology. Garland-STPM, New York, pp 111–118Google Scholar
  14. 14.
    Mitchison NA (1981a) Allospecific T cells. Cell Immunol 62:258–263CrossRefGoogle Scholar
  15. 15.
    Mitchison NA, (1981b) Information transfer between the minor antigen and T cell receptor repertoires. Scand J Immunol 14:631–635CrossRefGoogle Scholar
  16. 16.
    Mitchison NA, Kinlen L (1980) Present concepts in immune surveillance. In: Fougereau M, Dausset J (eds) Immunology 1980. Academic, London, pp 641–650Google Scholar
  17. 17.
    Mitchison NA, Lake P (1978) Latent help. In: Sercarz EE, Herzenberg LA, Fox CF (eds) Immune system: Genetics and regulation. ICN-UCLA Symposium Immune System, Park City, Utah, 1977. Academic, New York, pp 555–558Google Scholar
  18. 18.
    Simonsen M (1981) The major histocompatibility complex in a bird’s-eye view. In: Zaleski MB, Abeyounis CJ, Kano K (eds) Immunobiology of the major histocompatibility complex. 7th International Convocation of Immunology, 1980. Karger, Basel, pp 192–201Google Scholar
  19. 19.
    Simpson E, Matsunaga T, Brenan M, Brunner C, Benjamin D, Hetherington C, Hurme M, Chandler P (1980) H-Y antigen as a model for tumour antigens: the role of H-2-associative antigens in controlling anti-H-Y immune responses. Transplant Proc 12:103–106PubMedGoogle Scholar
  20. 20.
    Stern PL (1973) Theta alloantigen on mouse and rat fibroblasts. Nature 246:76–78Google Scholar
  21. 21.
    Von Boehmer H, Haas W (1979) Distinct Ir genes for helper and killer cells in the cytotoxic response to H-Y antigen. J Exp Med 150:1134–1142CrossRefGoogle Scholar
  22. 22.
    Wachtel SS, Ohno S (1979) The immunogenetics of sexual development. Prog Med Genet 3:109–142PubMedCrossRefGoogle Scholar
  23. 23.
    Williams AF, Gagnon J (1982) Neuronal cell thy-1 glycoprotein: homology with immunoglobulin. Science 216:696–702PubMedCrossRefGoogle Scholar
  24. 24.
    Yeh Ming, Czitrom AA, Mitchison NA (1982) Allospecific T-cell lines with functional activities. Immunology 46:281–287Google Scholar
  25. 25.
    Zaleski MB, Gorzynski TJ (1981) The Ir phenomenon and the MHC restriction: differences and similarities. In: Zaleski MB, Abeyounis CJ, Kano K (eds) Immunobiology of the major histocompatibility complex. 7th International Convocation of Immunology. Karger, Basel, pp 98–107Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1983

Authors and Affiliations

  • A. Fortunato
  • R. F. L. James
  • A. Mellor
  • N. A. Mitchison

There are no affiliations available

Personalised recommendations