The Journal of Immunology, Vol 154, Issue 2 762-771, Copyright © 1995 by American Association of Immunologists

ARTICLES

Generation of tumor-specific CTLs from melanoma patients by using peripheral blood stimulated with allogeneic melanoma tumor cell lines. Fine specificity and MART-1 melanoma antigen recognition

EJ Stevens, L Jacknin, PF Robbins, Y Kawakami, M el Gamil, SA Rosenberg and JR Yannelli
Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.

PBLs were isolated from 13 patients with metastatic melanoma. Mixed lymphocyte tumor cell cultures (ML TCs) were established (15 times) by using irradiated HLA-matched (one class I locus) allogeneic melanoma tumor cell lines (13 times) or autologous melanoma tumor cell lines (two times) in medium containing 120 IU/ml IL-2 and 100 IU/ml IL-4. PBLs grew to levels that could be assessed for functional reactivity 9 of 15 times. In seven of nine cases, CD3+CD8+ CTLs grew from MLTCs that were tumor specific; five were restricted by HLA-A2 and two were restricted by HLA-A24. Four of the tumor-specific CTL lines lysed autologous fresh tumor cells. Tumor-specific CTLs from two of three patients had cytolytic activity identical with tumor-infiltrating lymphocytes (TIL) derived from tumor biopsies removed earlier and grown in high concentrations (6000 IU/ml) of IL-2. Three of the HLA-A2- restricted tumor-specific CTLs were shown to recognize 293 cells transfected with HLA-A2.1 cDNA and the gene encoding the melanoma Ag, MART-1. In addition, these CTLs recognized the T2 cell line pulsed exogenously with the peptide MART-1(27-35), which is the nine-amino acid immunodominant epitope of the MART-1 Ag recognized on melanoma tumor cells by nearly all HLA-A2-restricted TIL. Thus, we have demonstrated the ability to generate tumor-specific CTLs from PBLs that are similar in their reactivity to TIL. This technique obviates the need for autologous tumor tissue and suggests that PBLs contain sufficient CTL precursors for use in generating antitumor CTLs for cellular immunotherapy trials.

This article has been cited by other articles:

				T. Zhang, B. A. Lemoi, and C. L. Sentman Chimeric NK-receptor-bearing T cells mediate antitumor immunotherapy Blood, September 1, 2005; 106(5): 1544 - 1551. [Abstract] [Full Text] [PDF]

				J. J. Roszkowski, G. E. Lyons, W. M. Kast, C. Yee, K. Van Besien, and M. I. Nishimura Simultaneous Generation of CD8+ and CD4+ Melanoma-Reactive T Cells by Retroviral-Mediated Transfer of a Single T-Cell Receptor Cancer Res., February 15, 2005; 65(4): 1570 - 1576. [Abstract] [Full Text] [PDF]

				M. Salio, D. Shepherd, P. R. Dunbar, M. Palmowski, K. Murphy, L. Wu, and V. Cerundolo Mature Dendritic Cells Prime Functionally Superior Melan-A-Specific CD8+ Lymphocytes as Compared with Nonprofessional APC J. Immunol., August 1, 2001; 167(3): 1188 - 1197. [Abstract] [Full Text] [PDF]

				J. T. Kurnick, T. Ramirez-Montagut, L. A. Boyle, D. M. Andrews, F. Pandolfi, P. J. Durda, D. Butera, I. S. Dunn, E. M. Benson, S. J. P. Gobin, et al. A Novel Autocrine Pathway of Tumor Escape from Immune Recognition: Melanoma Cell Lines Produce a Soluble Protein That Diminishes Expression of the Gene Encoding the Melanocyte Lineage Melan-A/MART-1 Antigen Through Down-Modulation of Its Promoter J. Immunol., August 1, 2001; 167(3): 1204 - 1211. [Abstract] [Full Text] [PDF]

				P. R. Dunbar, C. L. Smith, D. Chao, M. Salio, D. Shepherd, F. Mirza, M. Lipp, A. Lanzavecchia, F. Sallusto, A. Evans, et al. A Shift in the Phenotype of Melan-A-Specific CTL Identifies Melanoma Patients with an Active Tumor-Specific Immune Response J. Immunol., December 1, 2000; 165(11): 6644 - 6652. [Abstract] [Full Text] [PDF]

				N. Gervois, N. Labarriere, S. Le Guiner, M.-C. Pandolfino, J.-F. Fonteneau, Y. Guilloux, E. Diez, B. Dreno, and F. Jotereau High Avidity Melanoma-reactive Cytotoxic T Lymphocytes Are Efficiently Induced from Peripheral Blood Lymphocytes on Stimulation by Peptide-pulsed Melanoma Cells Clin. Cancer Res., April 1, 2000; 6(4): 1459 - 1467. [Abstract] [Full Text]

				H. M. Zarour, J. M. Kirkwood, L. S. Kierstead, W. Herr, V. Brusic, C. L. Slingluff Jr., J. Sidney, A. Sette, and W. J. Storkus Melan-A/MART-151-73 represents an immunogenic HLA-DR4-restricted epitope recognized by melanoma-reactive CD4+ T cells PNAS, January 4, 2000; 97(1): 400 - 405. [Abstract] [Full Text] [PDF]

				F. Wang, E. Bade, C. Kuniyoshi, L. Spears, G. Jeffery, V. Marty, S. Groshen, and J. Weber Phase I Trial of a MART-1 Peptide Vaccine with Incomplete Freund's Adjuvant for Resected High-Risk Melanoma Clin. Cancer Res., October 1, 1999; 5(10): 2756 - 2765. [Abstract] [Full Text] [PDF]

				T. M. Clay, M. C. Custer, J. Sachs, P. Hwu, S. A. Rosenberg, and M. I. Nishimura Efficient Transfer of a Tumor Antigen-Reactive TCR to Human Peripheral Blood Lymphocytes Confers Anti-Tumor Reactivity J. Immunol., July 1, 1999; 163(1): 507 - 513. [Abstract] [Full Text] [PDF]

				Y. Men, I. Miconnet, D. Valmori, D. Rimoldi, J.-C. Cerottini, and P. Romero Assessment of Immunogenicity of Human Melan-A Peptide Analogues in HLA-A*0201/Kb Transgenic Mice J. Immunol., March 15, 1999; 162(6): 3566 - 3573. [Abstract] [Full Text] [PDF]

				D. Valmori, N. Gervois, D. Rimoldi, J.-F. Fonteneau, A. Bonelo, D. Lienard, L. Rivoltini, F. Jotereau, J.-C. Cerottini, and P. Romero Diversity of the Fine Specificity Displayed by HLA-A*0201-Restricted CTL Specific for the Immunodominant Melan-A/MART-1 Antigenic Peptide J. Immunol., December 15, 1998; 161(12): 6956 - 6962. [Abstract] [Full Text] [PDF]

				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]