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Selective Loss of Human Leukocyte Class I Allospecificities and Staining of Melanoma Cells by Monoclonal Antibodies Recognizing Monomorphic Determinants of Class I Human Leukocyte Antigens¹

Department of Dermatology, Kumamoto University Medical School, Kumamoto, Japan [T. Ka., A. Y., T. Ki., T. O.]; Department of Microbiology and Immunology, New York Medical College, Valhalla, New York 10595 [Z. W., S. F.]; and Department of Radiation Oncology, New England Medical Center, Boston, Massachusetts [L. C., S. G-C.]

Immunoperoxidase staining of frozen sections from surgically removed melanoma lesions showed that anti-human leukocyte antigen (HLA)-A2, A28 monoclonal antibody (mAb) stained keratinocytes, but did not stain melanoma cells in 21% of the 14 primary and 44% of the 9 metastatic lesions tested. The loss of HLA-A2 and/or A28 allospecificities did not affect the staining patterns with mAb recognizing monomorphic determinants of HLA Class I antigens, in terms of percentage of stained melanoma cells and intensity of staining. This finding is not likely to reflect the sensitivity of the immunoperoxidase technique, since cytofluorographic analysis detected no significant difference in the staining pattern by mAb to monomorphic determinants of HLA Class I antigens between a melanoma cell line and an autologous transfectant that had acquired HLA-A2 antigens following gene transfer. The results of the present study imply that the frequency of abnormalities in HLA Class I antigen expression by melanoma cells is higher than that described in the literature, since selective losses of HLA Class I allospecificities are not detected by staining of melanoma cells with mAb to monomorphic determinants of HLA Class I antigens. The latter reagents have been used in most of the published studies to characterize the expression of HLA Class I antigens in melanoma lesions. Furthermore, the present results provide a mechanism for the unexpected resistance to cytotoxic T-cell-mediated lysis and the unexpected poor clinical course of the disease in some patients despite a high expression of HLA Class I antigens as measured by staining of melanoma cells with mAb to monomorphic determinants.

¹ This investigation was supported by USPHS Grants CA39559 and CA51814 awarded by the National Cancer Institute, Department of Health and Human Services. L. C. was supported by an AIRC fellowship.

² To whom requests for reprints should be addressed, at Department of Microbiology and Immunology, New York Medical College, Valhalla, NY 10595.

Received 12/31/92. Accepted 5/ 6/93.

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