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Expression of Melanoma-associated Antigens in Rapidly Dividing Human Melanocytes in Culture¹

The Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104

Conditions were established to induce rapid clonal growth of melanocytes from newborn foreskin. Surface antigen expression was analyzed using monoclonal antibodies derived by immunization of mice with melanoma cell, melanocyte, and placental membrane preparations. Unlike resting melanocytes in normal skin, cultured melanocytes expressed most major melanoma-associated antigens tested, e.g., nerve growth factor receptor, proteoglycan, transferrin-related M_r 97,000 protein antigen, M_r 120,000 protein, and gangliosides 9-O-acetyl G_D3 and G_D3. HLA-DR antigen and ganglioside G_D2 were expressed at very low levels or not expressed. After several subpassages, most melanocyte cultures, including clones and melanocytes, initially sorted by rosetting with monoclonal antibody to nerve growth factor receptor, lost their characteristic bipolar morphology and expression of nerve growth factor receptor and M_r 97,000 antigen but continued to express high molecular weight proteins such as proteoglycan, M_r 130,000/105,000 and 120,000 antigen. The few melanocyte cultures that did maintain their characteristic bipolar to spindle morphology continued to express all melanoma-associated antigens and even began to express HLA-DR antigens. Melanocytes cultured in the presence of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate also maintained their bipolar morphology, were often pigmented, and continued to express melanoma-associated antigens for several passages; they did not express HLA-DR antigen. Our studies indicate that rapidly proliferating melanocytes in culture undergo antigenic changes associated with malignancy.

¹ These studies were supported by grants from the NIH, CA-25874, CA-10815, and CA-29200, and by Grant IM-402 of the American Cancer Society.

² To whom requests for reprints should be addressed, at The Wistar Institute, 36th Street at Spruce, Philadelphia, PA 19104.

Received 6/30/86. Revised 10/23/86. Revised 3/ 5/87. Accepted 3/10/87.

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