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Effects of H-2K^b Gene on Expression of Melanoma-associated Antigen and Lectinbinding Sites on BL6 Melanoma Cells¹

Pittsburgh Cancer Institute and Department of Pathology, University of Pittsburgh, Pittsburgh, Pennsylvania 15213 [E. G., M. K., A. D., J. P. M.]; Laboratory of Virology, American Red Cross, Rockville, Maryland 20855 [G. J.]; and Laboratory of Cell Biology, National Cancer Institute, Bethesda, Maryland 20892 [V. H.]

An H-2K^b negative BL6 melanoma clone (BL6–8) was transfected with plasmids containing either the class I H-2K^b or class II H-2IA^k gene in combination with the neo^r gene. The effects of the transfected genes on the expression of the melanoma-associated antigen (MAA) recognized by the monoclonal antibodies MM2-9B6 and MM2-3C6 and the cell surface carbohydrates recognized by 15 different lectins were studied. The original H-2K^b- clone or clones transfected with neo^r or class II H-2IA^k genes expressed high levels of MAA and very low levels of soybean agglutinin (SBA), Griffonia simplicifolia I-B₄ (GSIB₄), and peanut agglutinin (PNA) lectin-binding sites. In contrast, clones that expressed high levels of the transfected H-2K^b gene completely lost the expression of MAA. In addition, these clones were characterized by the appearance of high levels of expression of the sugars specifically reacting with SBA, GSIB₄, and PNA lectins. When the original BL6–8 clone was transfected with the H-2K^d gene, 25 clones subsequently isolated had relatively low expression of the transfected H-2K^d gene but high expression of the endogenous H-2K^b gene accompanied by an alteration in expression of the MAA and lectin binding identical with patterns common for H-2K^b+ melanoma cells. These changes were not due to the transfection, plasmid construction, or place of insertion, since similar phenotypic characteristics were found in H-2K^b+ but not H-2K^b- clones isolated from the N-methyl-N'-nitro-N-nitrosoguanidine-treated BL6T2 or parental BL6 melanoma lines. In total, 73 BL6 melanoma clones were investigated and all of the 41 H-2K^b+ clones displayed loss of MAA and appearance of SBA, GSIB₄, and PNA-binding sugars. None of the 32 H-2K^b- clones showed these changes.

This study indicates that the class I H-2K^b gene product might alter several phenotypic properties of BL6 melanoma cells. The mechanisms of these changes remain unknown. We consider that these effects of the class I H-2K^b gene are indirect, involving interactions with the B-tropic ecotropic retrovirus specific for melanomas of C57BL/6 mice origin.

¹ This study was supported by Grant IM 581 from the American Cancer Society.

² To whom requests for reprints should be addressed, at Pittsburgh Cancer Institute, Biomedical Science Tower, W954, O'Hara & De Soto Sts., Pittsburgh, PA 15213.

Received 3/22/91. Accepted 7/17/91.

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