Functional consequence of variation in melanoma antigen expression |
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Authors: | Lee M Zehngebot Michael A Alexander DuPont Guerry IV Douglas B Cines Kenneth Mitchell and Meenhard Herlyn |
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Institution: | (1) Cancer Center, Hospital of the University of Pennsylvania, 19104 Philadelphia, PA, USA;(2) Glenolden Laboratory, E. I. DuPont de Nemours & Co. (Inc.), 500 South Ridgeway Avenue, 19036 Glenolden, PA, USA;(3) Wistar Institute, 19104 Philadelphia, PA, USA;(4) Present address: Division of Oncology, Albany Medical College, 12208 Albany, NY, USA |
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Abstract: | Summary Melanoma cells have been shown to express melanoma-associated antigens and, in many cases, the histocompatibility antigen, HLA-DR. We questioned whether the expression of these antigens was quantitatively altered during the serial passage of melanoma cells in culture. Therefore, we measured the binding of monoclonal antibodies specific for a melanoma-specific antigen and the HLA-DR antigen to melanoma cells from serial passages. Three cell lines were studied. We found that although both the melanoma-associated antigen and the HLA-DR antigen were qualitatively conserved, significant quantitative differences were seen. To study the functional consequences of these differences, we used fluorescence-activated cell sorting to create DR-enriched and DR-depleted populations from a single melanoma cell line heterogeneous for DR expression. We found that the proliferation of allogeneic T cells (measured by the 3H-TdR uptake) cultured with the DR-enriched and -depleted melanoma cell populations was directly related to the amount of the HLA-DR antigen expressed. These results indicate that in performance of experiments using melanoma cell lines quantitative assessment of antigenic expression is important, particularly if the function of a specific antigen is under examination. Further, our data clearly identify the HLA-DR antigen on melanoma cells as a participant in allogeneic lymphocyte stimulation.
Abbreviations used are: FACS, fluorescence activated cell sorter; FITC, fluorescein isothocyante; 3H-TdR, tritiated thymidine |
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