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The Journal of Immunology, Vol 154, Issue 5 2257-2265, Copyright © 1995 by American Association of Immunologists


ARTICLES

Induction of tumor-reactive CTL from peripheral blood and tumor- infiltrating lymphocytes of melanoma patients by in vitro stimulation with an immunodominant peptide of the human melanoma antigen MART-1

L Rivoltini, Y Kawakami, K Sakaguchi, S Southwood, A Sette, PF Robbins, FM Marincola, ML Salgaller, JR Yannelli and E Appella
Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.

MART-1 is an Ag expressed on melanomas and melanocytes, and is recognized by the majority of HLA-A2-restricted tumor-specific tumor- infiltrating lymphocytes (TIL) from melanoma patients. In the present study we have analyzed 10 potential 9-mer epitopes containing the HLA- A2.1 binding motifs for their ability to induce melanoma-specific T cell lines. Antimelanoma CTL could be generated only with MART-1(27-35) peptide, which has been previously shown to be recognized by a majority of HLA-A2-restricted TIL. Anti-MART-1(35-43)-specific CTL could also be induced, but these T cells did not recognize melanoma cells. MART-1(27- 35)-specific CTL could be effectively generated from a total of 11 of 12 PBL and from 3 of 3 TIL derived from HLA-A2+ melanoma patients, as well as from 2 of 4 PBL from HLA-A2+ healthy donors by in vitro stimulation with autologous PBMC pulsed with the synthetic MART-1(27- 35) peptide. These CTL lines specifically lysed and release cytokines (TNF-alpha, IFN-gamma, and GM-CSF) in response to T2 cells pulsed with MART-1(27-35), as well as to HLA-A2+ MART-1+ melanoma cells. CTL generated with MART-1(27-35) also lysed uncultured HLA-A2+ melanoma cells derived from tumor biopsies, indicating that this MART-1 epitope is likely to be expressed in association with HLA-A2 on the surface of tumor cells in vivo. CTL lines generated with MART-1(27-35) mediated 25- to 100-fold higher lytic activity than MART-1-reactive CTL grown from TIL in the presence of high dose IL-2. These results demonstrate that MART-1(27-35) peptide may represent an ideal candidate for Ag-specific immunotherapy in melanoma patients.


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M. P. Bettinotti, C. J. Kim, K.-H. Lee, M. Roden, J. N. Cormier, M. Panelli, K. K. Parker, and F. M. Marincola
Stringent Allele/Epitope Requirements for MART-1/Melan A Immunodominance: Implications for Peptide-Based Immunotherapy
J. Immunol., July 15, 1998; 161(2): 877 - 889.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
K. F. Yoong, G. McNab, S. G. Hubscher, and D. H. Adams
Vascular Adhesion Protein-1 and ICAM-1 Support the Adhesion of Tumor-Infiltrating Lymphocytes to Tumor Endothelium in Human Hepatocellular Carcinoma
J. Immunol., April 15, 1998; 160(8): 3978 - 3988.
[Abstract] [Full Text] [PDF]


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