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Induction of Melanoma Reactive T Cells by Stimulator Cells Expressing Melanoma Epitope-Major Histocompatibility Complex Class I Fusion Proteins

Surgery Branch, Division of Clinical Sciences, National Cancer Institute, NIH, Bethesda, MD 20892

Several epitopes in the human melanoma antigens recognized by HLA-A2-restricted CTLs have a relatively low MHC-binding affinity and as a result may be expressed at very low densities on the cell surface, indicating that these epitopes may not be efficient immunogens. To express these epitopes at higher densities on the surface of antigen-presenting cells and therefore improve their immunogenicity, a DNA construct in which a cDNA fragment encoding the melanoma epitope MART-1_27–35 or gp100_280–288 was inserted between sequences encoding the leader and the HLA-A*0201 protein. Cells transfected with these epitope-HLA fusion constructs were recognized by HLA-A2-restricted melanoma-reactive CTLs specific for the MART-1 or gp100 epitope. In addition, tumor-reactive CTLs could be induced from PBMCs of patients with metastatic melanoma by in vitro stimulation with HMY-C1R B-cell lines expressing the MART-1 or gp100 epitope-HLA-A*0201 fusion protein. These epitope-HLA fusion constructs may be useful for the development of immunotherapies for patients with melanoma.

¹ To whom requests for reprints should be addressed, at Surgery Branch, Division of Clinical Sciences, Building 10, Room 2B42, 10 Center Drive, MSC 1502, National Cancer Institute, NIH, Bethesda, MD 20892.

Received 11/ 5/96. Accepted 12/ 3/96.

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