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Generation of human-melanoma-specific T lymphocyte clones defining novel cytolytic targets with panels of newly established melanoma cell lines

  • Original Article
  • Melanoma, Melanoma-Associated Antigens, Cytolytic T Cell Clones, HLA-A2, Immunogenicity
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Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Melanoma is a cancer where the immune system is believed to play an important role in the control of malignant cell growth. To study the variability of the immune response in melanoma patients, we derived melanoma cell lines from several HLA-A2+ and HLA-A2− patients. The melanoma cell lines studied were designated FM3, FM6, FM9, FM28, FM37, FM45, FM55P, FM55M1 and FM55M2 and were established from eight metastatic tumors as well as from one primary tumor from a total of seven different patients. On the basis of the ability of tumor cells to induce specific cytotoxic T lymphocytes (CTL) from peripheral blood lymphocytes (PBL) in mixed lymphocyte/tumor culture with HLA-A2+ melanoma cells, the FM3 cell line was characterized as highly immunogenic. To investigate the expression of different melanoma-associated antigens recognized by CTL on different melanoma cell lines, we selected the cell line FM3 for restimulation and further T cell cloning experiments. The lytic activity of CTL clones with good proliferative activity was examined using a panel of HLA-A2+ and HLA-A2− melanoma cell lines. None of the tested HLA-A2− melanoma cell lines were susceptible to lysis by the CTL clones, whereas allogeneic HLA-A2+ melanoma cell lines were lysed only by a few CTL clones. On the basis of their reactivity with different melanoma cell lines, it was possible to divide the present CTL clones into at least four groups suggesting the recognition of at least four different antigens. Three of these target structures probably are different from already-described HLA-A2-restricted melanoma-associated antigens, because their expression in the different melanoma cell lines do not correlate with the recognition of melanoma cells by these CTL. The results first indicate that poorly immunogenic melanoma cells may express melanoma-associated antigens, and also suggest that, by using CTL clones obtained against different HLA-class-I-matched melanoma cells, it is possible to define such antigens.

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References

  1. Altomonte M, Gloghini A, Bertola G, Gasparollo A, Carbone A, Ferrone S, Maio M (1993) Differential expression of cell adhesion molecules CD54/CD11a and CD58/CD2 by human melanoma cells and functional role in their interaction with cytotoxic cells. Cancer Res 53:3343

    PubMed  Google Scholar 

  2. Anichini A, Fossati G, Parmiani G (1985) Clonal analysis of cytotoxic T-lymphocytes response to autologous human metastatic melanoma. Int J Cancer 35:683

    PubMed  Google Scholar 

  3. Anichini A, Maccalli C, Mortarini R, Salvi S, Mazzocchi A, Squarcina P, Herlyn M, Parmiani G (1993) Melanoma cells and normal melanocytes share antigens recognized by HLA-A2-restricted cytotoxic T cell clones from melanoma patients. J Exp Med 177:989

    PubMed  Google Scholar 

  4. Anichini A, Mortarini R, Alberti S, Mantovani A, Parmiani G (1993) T-cell-receptor engagement and tumor ICAM-1 upregulation are required to by-pass low susceptibility of melanoma cells to autologous CTL-mediated lysis. Int J Cancer 53:994

    PubMed  Google Scholar 

  5. Bakker ABH, Schreurs MWJ, Boer AJ de, Kawakami Y, Rosenberg SA, Adema GJ, Figdor CG (1994) Melanocyte lineagespecific antigen gp100 is recognized by melanoma-derived tumor-infiltrating lymphocytes. J Exp Med 179:1005

    PubMed  Google Scholar 

  6. Becker JC, Brabletz T, Czerny C, Termeer C, Bröcker EB (1993) Tumor escape mechanisms from immunosurveillance-induction of unresponsiveness in a specific MHC-restricted CD4+ human T-cell clone by the autologous MHC class-II+ melanoma. Int Immunol 5:1501

    PubMed  Google Scholar 

  7. Becker JC, Schwinn A, Dummer R, Burg G, Bröcker EB (1993) Lesion-specific activation of cloned human tumor-infiltrating lymphocytes by autologous tumor cells: induction of proliferation and cytokine production. J Invest Dermatol 101:15

    PubMed  Google Scholar 

  8. Benathan M, Pararas C, Frenk E (1991) Modulatory growth effects of 3T3 fibroblasts on cocultivated human melanoma cells. Anticancer Res 11:203

    PubMed  Google Scholar 

  9. Bodmer H, Ogg G, Cotch F, McMichael A (1989) Anti-HLA-A antibody-enhancement of peptide association with HLA-A2 as detected by cytotoxic T lymphocytes. Nature 342:443

    PubMed  Google Scholar 

  10. Boël P, Wildman C, Sensi ML, Brasseur R, Renauld J-C, Coulie P, Boon T, Van der Bruggen P (1985) BAGE: a new gene encoding an antigen recognized on human melanomas by cytolytic T lymphocytes. Immunity 2:167

    Google Scholar 

  11. Brichard V, Van Pel A, Wölfel T, Wölfel C, De Plaen E, Lethé B, Coulie P, Boon T (1993) The tyrosinase gene codes for an antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J Exp Med 178:489

    PubMed  Google Scholar 

  12. Browning MJ, Krausa P, Rowan A, Bicknell DC, Bodmer JG, Bodmer WF (1993) Tissue typing of the HLA-A from genomic DNA by sequence specific PCR: Comparison of HLA genotype genotype and surface expression on colorectal tumor lines. Proc Natl Acad Sci USA 90:2842

    PubMed  Google Scholar 

  13. Castelli C, Storkus WJ, Maeurer MJ, Martin DM, Huang EC, Pramanik BN, Nagabhushan TL, Parmiani G, Lotze MT (1995) Mass spectrometric identification of a naturally processed melanoma peptide recognized by CD8(+) cytotoxic T lymphocytes. J Exp Med 181:363

    PubMed  Google Scholar 

  14. Chen Q, Hersey P (1992) MHC-restricted responses of CD8+ and CD4+ T-cell clones from regional lymph nodes of melanoma patients. Int J Cancer 51:218

    PubMed  Google Scholar 

  15. Chen LP, McGowan P, Ashe S, Johnston J, Li YW, Hellström I, Hellström KE (1994) Tumor immunogenicity determines the effect of B7 costimulation on T cell-mediated tumor immunity. J Exp Med 179:523

    PubMed  Google Scholar 

  16. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156

    PubMed  Google Scholar 

  17. Coulie PG, Somville M, Lehmann F, Hainaut P, Brasseur F, Devos R, Roon T (1992) Precursor frequency analysis of human cytolytic T lymphocytes directed against autologous melanoma cells. Int J Cancer 50:289

    PubMed  Google Scholar 

  18. Coulie PG, Brichard V, Van Pel A, Wölfel T, Schneider J, Traversari C, Mattei S, De Plaën E, Lurquin C, Szikora J-P, Renauld J-C, Boon T (1994) A new gene coding for a differentiation antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J Exp Med 180:35

    PubMed  Google Scholar 

  19. Crowley NJ, Slingluff CL, Darrow TL, Seigler HF (1990) Generation of human autologous melanoma-specific cytotoxic T-cells using HLA-A2 matched allogeneic melanomas. Cancer Res 50:492

    PubMed  Google Scholar 

  20. Crowley NJ, Darrow TL, Quinn-Allen MA, Seigler HF (1991) MHC-restricted recognition of autologous melanoma by tumor-specific cytotoxic T cells. Evidence for restriction by a dominant HLA-A allele. J Immunol 146:1692

    PubMed  Google Scholar 

  21. Darrow TL, Slingluff CL, Seigler HF (1989) The role of HLA class I antigens in recognition of melanoma cells by tumorspecific cytotoxic T lymphocytes. J Immunol 142:3329

    PubMed  Google Scholar 

  22. Gaugler B, Van den Eynde B, Van der Bruggen P, Romero P, Gaforio JJ, De Olaen E, Lethé B, Brasseur F, Boon T (1994) Human gene MAGE-3 codes for an antigen recognized on a melanoma by autologous cytolytic T lymphocytes. J Exp Med 179:921

    PubMed  Google Scholar 

  23. Hérin M, Lemoine C, Weynants P, Vessière F, Knuth A, Devos R, Boon T (1987) Production of stable cytolytic T-cell clones directed against autologous human melanoma. Int J Cancer 39:390

    PubMed  Google Scholar 

  24. Hermann GG, Zeuthen J, Claesson MH (1992) LAK cell mediated cytotoxicity against tumor cell targets used to monitor the stimulatory effect of interleukin-2. Cytotoxicity, target cell recognition and phenotype of effector cells lysing the Daudi, T24 and K562 tumor cell lines. Nat Immun Cell Growth Regul 11:7

    Google Scholar 

  25. Hersey P, Si ZY, Smith MJ, Thomas WD (1994) Expression of the co-stimulatory molecule B7 on melanoma cells. Int J Cancer 58:527

    PubMed  Google Scholar 

  26. Itoh K, Platsoucas CD, Balch CM (1988) Autologous tumorspecific cytotoxic T-lymphocytes in the infiltrate of human metastatic melanomas: activation by interleukin-2 and autologous tumor cells, and involvement of the T cell receptor. J Exp Med 168:1419

    PubMed  Google Scholar 

  27. Johnston RF, Pickett SC, Barker DL (1990) Autoradiography using storage phosphor technology. Electrophoresis 11:355

    PubMed  Google Scholar 

  28. Kawakami Y, Eliyahu S, Delgado CH, Robbins PF, Rivoltini L, Topalian SL, Miki T Rosenberg SA (1994) Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor. Proc Natl Acad Sci USA 91:3515

    PubMed  Google Scholar 

  29. Kawakami Y, Eliyahu S, Delgado CH, Robbins PH, Sakaguchi K, Appella E, Yannelli JH, Adema GH, Miki T, Rosenberg SA (1994) Identification of a human melanoma antigen recognized by tumor-infiltrating lymphocytes associated with in vivo tumor rejection. Proc Natl Acad Sci USA 91:6458

    PubMed  Google Scholar 

  30. Knuth A, Wölfel T, Klehmann E, Boon T, Meyer zum Büschenfelde KH (1989) Cytolytic T-cell clones against an autologous human melanoma: specificity study and definition of three antigens by immunoselection. Proc Natl Acad Sci USA 86:2804

    PubMed  Google Scholar 

  31. Kwon BS, Haq AK, Pomerantz SH, Halaban R (1987) Isolation and sequence of a cDNA clone for human tyrosinase that maps at the mouse c-albino locus. Proc Natl Acad Sci USA 84:7473

    PubMed  Google Scholar 

  32. Muul LM, Spiess PJ, Director EP, Rosenberg SA (1987) Identification of specific cytolytic immune response against autologous tumor in humans bearing malignant melanoma. J Immunol 138:989

    PubMed  Google Scholar 

  33. Olsen AC, Fossum B, Kirkin AF, Zeuthen J, Gaudernack G (1995) A human melanoma cell line, recognized by both HLA class I and class II restricted T cells, is capable of initiating both primary and secondary responses. Scand J Immunol 41:357

    PubMed  Google Scholar 

  34. O'Neil BH, Kawakami Y, Restifo NP, Bennink JR, Yewdell JW, Rosenberg SA (1993) Detection of shared MHC-restricted human melanoma antigens after vaccinia virus-mediated transduction of genes coding for HLA. J Immunol 151:1410

    PubMed  Google Scholar 

  35. Ramarathinam L, Castle M, Wu Y, Liu Y (1994) T-cell costimulation by B7/BB1 induces CD8 T-cell-dependent tumor rejection — an important role of B7/BB1 in the induction, recruitment, and effector function of anti-tumor T-cells. J Exp Med 179:1205

    PubMed  Google Scholar 

  36. Schwartz RH (1989) Acquisition of immunological self-tolerance. Cell 57:1073

    PubMed  Google Scholar 

  37. Topalian S, Solomon D, Rosenberg SA (1989) Tumor-specific cytolysis by lymphocytes infiltrating human melanomas. J Immunol 142:3714

    PubMed  Google Scholar 

  38. Topalian SL, Hom SS, Kawakami Y, Mancini M, Schwartzentruber DJ, Zakut R, Rosenberg SA (1992) Recognition of shared melanoma antigens by human tumor-infiltrating lymphocytes. J Immunother 12:203

    PubMed  Google Scholar 

  39. Townsend SE, Allison JP (1993) Tumor rejection after direct costimulation of CD8+ T cells by B7-transfected melanoma cells. Science 259:368

    PubMed  Google Scholar 

  40. Van der Bruggen P, Traversari C, Chomez P, Lurquin C, De Plaen E, Van den Eynde B, Knuth A, Boon T (1991) A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science 254:1643

    PubMed  Google Scholar 

  41. Van der Bruggen P, Bastin J, Gajewski T, Coulie PG, Boël P, Desmet C, Traversari C, Townsend A, Boon T (1994) A peptide encoded by human gene MAGE-3 and presented by HLA-A2 induces cytolytic T lymphocytes that recognize tumor cells expressing MAGE-3. Eur J Immunol 24:3038

    PubMed  Google Scholar 

  42. Vessière-Louveaux F, Darville M, Knuth A, Boon T (1985) Use of irradiated mouse fibroblasts to improve the cloning and adaptation to culture of human melanoma cells. Int J Cancer 35:231

    PubMed  Google Scholar 

  43. Viret C, Davodeau F, Guilloux Y, Bignon JD, Semana G, Breathlach R, Jotereau F (1993) Recognition of shared melanoma antigen by HLA-A2-restricted cytolytic T cell clones derived from human tumor-infiltrating lymphocytes. Eur J Immunol 23:141

    PubMed  Google Scholar 

  44. Wang RF, Robbins PF, Kawakami Y, Kang XQ, Rosenberg SA (1995) Identification of a gene encoding a melanoma tumor antigen recognized by HLA-A31-restricted tumor-infiltrating lymphocytes. J Exp Med 181:799

    PubMed  Google Scholar 

  45. Wölfel T, Klehmann E, Müller C, Schütt K-H, zum Büschenfelde KHM, Knuth A (1989) Lysis of human melanoma cells by an autologous cytolytic T cell clones. Identification of human histocompatibility leukocyte antigen A2 as a restriction element for three different antigens. J Exp Med 170:797

    PubMed  Google Scholar 

  46. Wölfel T, Van Pel A, Brichard V, Schneider J, Seliger B, zum Büschenfelde KHM, Boon T (1994) Two tyrosinase nonapeptides recognized on HLA-A2 melanomas by autologous cytolytic T lymphocytes. Eur J Immunol 24:759

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Tumor Cell Biology, Division of Cancer Biology, Danish Cancer Society, Strandboulevarden 49, Bldg. 7.1, DK-2100, Copenhagen, Denmark

    Alexei F. Kirkin, Troels Reichert Petersen, Anna Catharina Olsen, Li Li, Per thor Straten & Jesper Zeuthen

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  1. Alexei F. Kirkin
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  2. Troels Reichert Petersen
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  3. Anna Catharina Olsen
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  4. Li Li
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  5. Per thor Straten
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  6. Jesper Zeuthen
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Kirkin, A.F., Petersen, T.R., Olsen, A.C. et al. Generation of human-melanoma-specific T lymphocyte clones defining novel cytolytic targets with panels of newly established melanoma cell lines. Cancer Immunol Immunother 41, 71–81 (1995). https://doi.org/10.1007/BF01527402

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  • DOI: https://doi.org/10.1007/BF01527402

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