The Journal of Experimental Medicine
VeriKine-HS Human IFN-Beta
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© The Rockefeller University Press, 0022-1007/1997/2/453/ $5.00
The Journal of Experimental Medicine, Volume 185, Number 3, February 3, 1997 453-460

Articles

Identification of Tyrosinase-related Protein 2 as a Tumor Rejection Antigen for the B16 Melanoma

Matthew B. Bloom, Donna Perry-Lalley, Paul F. Robbins, Yong Li, Mona El-Gamil, Steven A. Rosenberg, and James C. Yang

From the Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892

Recently, major advances have been made in the identification of antigens from human melanoma which are recognized by T cells. In spite of this, little is known about the optimal ways to use these antigens to treat patients with cancer. Progress in this area is likely to require accurate preclinical animal models, but the availability of such models has lagged behind developments in human tumor immunology. Whereas many of the identified human melanoma antigens are normal tissue differentiation proteins, analogous murine tumor antigens have not yet been identified. In this paper we identify a normal tissue differentiation antigen, tyrosinaserelated protein 2 (TRP-2), expressed by the murine B16 melanoma which was found by screening a cDNA library from B16 with tumor-reactive cytotoxic T lymphocytes (CTL). A peptide conforming to the predicted MHC class I H2-Kb binding motif, TRP-2181-188, was identified as the major reactive epitope within TRP-2 recognized by these anti-B16 CTLs. By site-directed mutagenesis, it was shown that alteration of this epitope eliminated recognition of TRP-2. It was further demonstrated that a CTL line raised from splenocytes by repeated stimulation in vitro with this peptide could recognize B16 tumor and was therapeutic against 3-d-old established pulmonary metastases. The use of TRP-2 in a preclinical model of tumor immunotherapy may be helpful in suggesting optimal vaccination strategies for cancer therapy in patients.

Address correspondence to Dr. James C. Yang, Surgery Branch, National Cancer Institute, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892.

M. Bloom is a Howard Hughes Medical Institute–National Institutes of Health Research Scholar.

1Abbreviations used in this paper: CM, complete media; gal, galactosidase; MART, melanoma antigen reactive with T cells; TRP, tyrosinase-related protein.


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