Transfection and expression of a gene coding for a human-melanoma antigen recognized by autologous cytolytic T lymphocytes
(1992) Immunogenetics — Vol. 35, n° 3, p. 145-152 (1992)
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- Abstract
- Human melanoma line MZ2-MEL expresses several antigens recognized by autologous cytolytic T lymphocytes (CTL). As a first step towards the cloning of the gene coding for one of these antigens, we tried to obtain transfectants expressing the antigen. The DNA recipient cell was a variant of MZ2-MEL which had been selected with a CTL clone for the loss of antigen E. It was cotransfected with genomic DNA of the original melanoma line and with selective plasmid pSVtkneo-beta. Geneticin-resistant transfectants were obtained at a frequency of 2 x 10(-4). These transfectants were then screened for their ability to stimulate the production of tumor necrosis factor by the anti-E CTL clone. One transfectant expressing antigen E was identified among 70 000 drug-resistant transfectants. Its sensitivity to lysis by the anti-E CTL was equal to that of the original melanoma cell line. When this transfectant was submitted to immunoselection with the anti-E CTL clone, the resulting antigen-loss variants were found to have lost several of the transfected pSVtkneo-beta sequences. This indicated that the gene coding for the antigen had been integrated in the vicinity of pSVtkneo-beta sequences, as expected for cotransfected DNA.
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Citations
Traversari, C., van der Bruggen, P., Van den Eynde, B., Hainaut, P., Lemoine, C., Ohta, N., Old, L., & Boon, T. (1992). Transfection and expression of a gene coding for a human-melanoma antigen recognized by autologous cytolytic T lymphocytes. Immunogenetics, 35(3), 145-152. https://doi.org/10.1007/BF00185107 (Original work published 1992)