Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 Jun 11;93(12):5915–5919. doi: 10.1073/pnas.93.12.5915

Serological analysis of Melan-A(MART-1), a melanocyte-specific protein homogeneously expressed in human melanomas.

Y T Chen 1, E Stockert 1, A Jungbluth 1, S Tsang 1, K A Coplan 1, M J Scanlan 1, L J Old 1
PMCID: PMC39162  PMID: 8650193

Abstract

Recent progress in the structural identification of human melanoma antigens recognized by autologous cytotoxic T cells has led to the recognition of a new melanocyte differentiation antigen, Melan-A(MART-1). To determine the properties of the Melan-A gene product, Melan-A recombinant protein was produced in Escherichia coli and used to generate mouse monoclonal antibodies (mAbs). Two prototype mAbs, A103 and A355, were selected for detailed study. Immunoblotting results with A103 showed a 20-22-kDa doublet In Melan-A mRNA positive melanoma cell lines and no reactivity with Melan-A mRNA-negative cell lines. A355, in addition to the 20-22-kDa doublet, recognized several other protein species in Melan-A mRNA-positive cell lines. Immunocytochemical assays on cultured melanoma cells showed specific and uniform cytoplasmic staining in Melan-A mRNA-positive cell lines. Immunohistochemical analysis of normal human tissues with both mAbs showed staining of adult melanocytes and no reactivity with the other normal tissues tested. Analysis of 21 melanoma specimens showed homogenous staining of tumor cell cytoplasm in 16 of 17 Melan-A mRNA-positive cases and no reactivity with the three Melan-A mRNA-negative cases.

Full text

PDF
5915

Images in this article

5916Fig. 1
on p.5916
5916Fig. 2
on p.5916
5917Fig. 3
on p.5917

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boon T., Cerottini J. C., Van den Eynde B., van der Bruggen P., Van Pel A. Tumor antigens recognized by T lymphocytes. Annu Rev Immunol. 1994;12:337–365. doi: 10.1146/annurev.iy.12.040194.002005. [DOI] [PubMed] [Google Scholar]
  2. Boël P., Wildmann C., Sensi M. L., Brasseur R., Renauld J. C., Coulie P., Boon T., van der Bruggen P. BAGE: a new gene encoding an antigen recognized on human melanomas by cytolytic T lymphocytes. Immunity. 1995 Feb;2(2):167–175. doi: 10.1016/s1074-7613(95)80053-0. [DOI] [PubMed] [Google Scholar]
  3. Brasseur F., Marchand M., Vanwijck R., Hérin M., Lethé B., Chomez P., Boon T. Human gene MAGE-1, which codes for a tumor-rejection antigen, is expressed by some breast tumors. Int J Cancer. 1992 Nov 11;52(5):839–841. doi: 10.1002/ijc.2910520528. [DOI] [PubMed] [Google Scholar]
  4. Brichard V., Van Pel A., Wölfel T., Wölfel C., De Plaen E., Lethé B., Coulie P., Boon T. The tyrosinase gene codes for an antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J Exp Med. 1993 Aug 1;178(2):489–495. doi: 10.1084/jem.178.2.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carey T. E., Lloyd K. O., Takahashi T., Travassos L. R., Old L. J. AU cell-surface antigen of human malignant melanoma: solubilization and partial characterization. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2898–2902. doi: 10.1073/pnas.76.6.2898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chen Y. T., Stockert E., Chen Y., Garin-Chesa P., Rettig W. J., van der Bruggen P., Boon T., Old L. J. Identification of the MAGE-1 gene product by monoclonal and polyclonal antibodies. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):1004–1008. doi: 10.1073/pnas.91.3.1004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen Y. T., Stockert E., Tsang S., Coplan K. A., Old L. J. Immunophenotyping of melanomas for tyrosinase: implications for vaccine development. Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8125–8129. doi: 10.1073/pnas.92.18.8125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coulie P. G., Brichard V., Van Pel A., Wölfel T., Schneider J., Traversari C., Mattei S., De Plaen E., Lurquin C., Szikora J. P. A new gene coding for a differentiation antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J Exp Med. 1994 Jul 1;180(1):35–42. doi: 10.1084/jem.180.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dippold W. G., Lloyd K. O., Li L. T., Ikeda H., Oettgen H. F., Old L. J. Cell surface antigens of human malignant melanoma: definition of six antigenic systems with mouse monoclonal antibodies. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6114–6118. doi: 10.1073/pnas.77.10.6114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Garin-Chesa P., Fellinger E. J., Huvos A. G., Beresford H. R., Melamed M. R., Triche T. J., Rettig W. J. Immunohistochemical analysis of neural cell adhesion molecules. Differential expression in small round cell tumors of childhood and adolescence. Am J Pathol. 1991 Aug;139(2):275–286. [PMC free article] [PubMed] [Google Scholar]
  11. Gaugler B., Van den Eynde B., van der Bruggen P., Romero P., Gaforio J. J., De Plaen E., Lethé B., Brasseur F., Boon T. Human gene MAGE-3 codes for an antigen recognized on a melanoma by autologous cytolytic T lymphocytes. J Exp Med. 1994 Mar 1;179(3):921–930. doi: 10.1084/jem.179.3.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gown A. M., Vogel A. M., Hoak D., Gough F., McNutt M. A. Monoclonal antibodies specific for melanocytic tumors distinguish subpopulations of melanocytes. Am J Pathol. 1986 May;123(2):195–203. [PMC free article] [PubMed] [Google Scholar]
  13. Kawakami Y., Eliyahu S., Delgado C. H., Robbins P. F., Rivoltini L., Topalian S. L., Miki T., Rosenberg S. A. Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3515–3519. doi: 10.1073/pnas.91.9.3515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kawakami Y., Eliyahu S., Delgado C. H., Robbins P. F., Sakaguchi K., Appella E., Yannelli J. R., Adema G. J., Miki T., Rosenberg S. A. Identification of a human melanoma antigen recognized by tumor-infiltrating lymphocytes associated with in vivo tumor rejection. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6458–6462. doi: 10.1073/pnas.91.14.6458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kawakami Y., Eliyahu S., Sakaguchi K., Robbins P. F., Rivoltini L., Yannelli J. R., Appella E., Rosenberg S. A. Identification of the immunodominant peptides of the MART-1 human melanoma antigen recognized by the majority of HLA-A2-restricted tumor infiltrating lymphocytes. J Exp Med. 1994 Jul 1;180(1):347–352. doi: 10.1084/jem.180.1.347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kocher T., Schultz-Thater E., Gudat F., Schaefer C., Casorati G., Juretic A., Willimann T., Harder F., Heberer M., Spagnoli G. C. Identification and intracellular location of MAGE-3 gene product. Cancer Res. 1995 Jun 1;55(11):2236–2239. [PubMed] [Google Scholar]
  17. Kwon B. S. Pigmentation genes: the tyrosinase gene family and the pmel 17 gene family. J Invest Dermatol. 1993 Feb;100(2 Suppl):134S–140S. doi: 10.1111/1523-1747.ep12465022. [DOI] [PubMed] [Google Scholar]
  18. Rettig W. J., Chesa P. G., Beresford H. R., Feickert H. J., Jennings M. T., Cohen J., Oettgen H. F., Old L. J. Differential expression of cell surface antigens and glial fibrillary acidic protein in human astrocytoma subsets. Cancer Res. 1986 Dec;46(12 Pt 1):6406–6412. [PubMed] [Google Scholar]
  19. Rettig W. J., Garin-Chesa P., Healey J. H., Su S. L., Ozer H. L., Schwab M., Albino A. P., Old L. J. Regulation and heteromeric structure of the fibroblast activation protein in normal and transformed cells of mesenchymal and neuroectodermal origin. Cancer Res. 1993 Jul 15;53(14):3327–3335. [PubMed] [Google Scholar]
  20. Rettig W. J., Su S. L., Fortunato S. R., Scanlan M. J., Raj B. K., Garin-Chesa P., Healey J. H., Old L. J. Fibroblast activation protein: purification, epitope mapping and induction by growth factors. Int J Cancer. 1994 Aug 1;58(3):385–392. doi: 10.1002/ijc.2910580314. [DOI] [PubMed] [Google Scholar]
  21. Rimoldi D., Romero P., Carrel S. The human melanoma antigen-encoding gene, MAGE-1, is expressed by other tumour cells of neuroectodermal origin such as glioblastomas and neuroblastomas. Int J Cancer. 1993 May 28;54(3):527–528. doi: 10.1002/ijc.2910540329. [DOI] [PubMed] [Google Scholar]
  22. Robbins P. F., el-Gamil M., Kawakami Y., Stevens E., Yannelli J. R., Rosenberg S. A. Recognition of tyrosinase by tumor-infiltrating lymphocytes from a patient responding to immunotherapy. Cancer Res. 1994 Jun 15;54(12):3124–3126. [PubMed] [Google Scholar]
  23. Salvi S., Segalla F., Rao S., Arienti F., Sartori M., Bratina G., Caronni E., Anichini A., Clemente C., Parmiani G. Overexpression of the T-cell receptor beta-chain variable region TCRBV14 in HLA-A2-matched primary human melanomas. Cancer Res. 1995 Aug 1;55(15):3374–3379. [PubMed] [Google Scholar]
  24. Schultz-Thater E., Juretic A., Dellabona P., Lüscher U., Siegrist W., Harder F., Heberer M., Zuber M., Spagnoli G. C. MAGE-1 gene product is a cytoplasmic protein. Int J Cancer. 1994 Nov 1;59(3):435–439. doi: 10.1002/ijc.2910590324. [DOI] [PubMed] [Google Scholar]
  25. Sensi M., Traversari C., Radrizzani M., Salvi S., Maccalli C., Mortarini R., Rivoltini L., Farina C., Nicolini G., Wölfel T. Cytotoxic T-lymphocyte clones from different patients display limited T-cell-receptor variable-region gene usage in HLA-A2-restricted recognition of the melanoma antigen Melan-A/MART-1. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5674–5678. doi: 10.1073/pnas.92.12.5674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Thomson T. M., Mattes M. J., Roux L., Old L. J., Lloyd K. O. Pigmentation-associated glycoprotein of human melanomas and melanocytes: definition with a mouse monoclonal antibody. J Invest Dermatol. 1985 Aug;85(2):169–174. doi: 10.1111/1523-1747.ep12276608. [DOI] [PubMed] [Google Scholar]
  27. Van den Eynde B., Peeters O., De Backer O., Gaugler B., Lucas S., Boon T. A new family of genes coding for an antigen recognized by autologous cytolytic T lymphocytes on a human melanoma. J Exp Med. 1995 Sep 1;182(3):689–698. doi: 10.1084/jem.182.3.689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wang R. F., Robbins P. F., Kawakami Y., Kang X. Q., Rosenberg S. A. Identification of a gene encoding a melanoma tumor antigen recognized by HLA-A31-restricted tumor-infiltrating lymphocytes. J Exp Med. 1995 Feb 1;181(2):799–804. doi: 10.1084/jem.181.2.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. van der Bruggen P., Traversari C., Chomez P., Lurquin C., De Plaen E., Van den Eynde B., Knuth A., Boon T. A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science. 1991 Dec 13;254(5038):1643–1647. doi: 10.1126/science.1840703. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES