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. 1998 Jun 15;101(12):2720–2729. doi: 10.1172/JCI498

beta2-Microglobulin mutations, HLA class I antigen loss, and tumor progression in melanoma.

D J Hicklin 1, Z Wang 1, F Arienti 1, L Rivoltini 1, G Parmiani 1, S Ferrone 1
PMCID: PMC508863  PMID: 9637706

Abstract

The potential negative impact of HLA class I antigen abnormalities on the outcome of T cell-based immunotherapy of melanoma has prompted us to investigate the mechanisms underlying lack of HLA class I antigen expression by melanoma cell lines Me18105, Me9923, and Me1386. Distinct mutations in the beta2-microglobulin (beta2m) gene were identified in each cell line which result in loss of functional beta2m. In Me18105 cells, an aberrant splicing mechanism caused by an A--> G point mutation in the splice acceptor site of intron 1 of the beta2m gene, deletes 11 bp from the beta2m mRNA creating a shift in the reading frame. In Me9923 cells a 14-bp deletion in exon 2 and in Me1386 cells a CT deletion in exon 1 of the beta2m gene produce a frameshift mutation. The beta2m gene mutations identified in Me18105, Me9923, and Me1386 cells were also detected in the surgically removed melanoma lesions from which the cell lines originated. Transfection of each melanoma cell line with a wild-type beta2m gene restored HLA class I antigen expression and, in Me18105 cells, recognition by Melan-A/MART-1-specific, HLA-A2-restricted cytotoxic T lymphocytes. Interestingly, the beta2m mutation present in Me9923 cells that were derived from a metastatic lesion was also found in the Me9923P cell line that originated from the autologous primary lesion. These data suggest that beta2m mutations in melanoma cells may be an early event in progression to the malignant phenotype.

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Selected References

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

  1. Anichini A., Maccalli C., Mortarini R., Salvi S., Mazzocchi A., Squarcina P., Herlyn M., Parmiani G. Melanoma cells and normal melanocytes share antigens recognized by HLA-A2-restricted cytotoxic T cell clones from melanoma patients. J Exp Med. 1993 Apr 1;177(4):989–998. doi: 10.1084/jem.177.4.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barnstable C. J., Bodmer W. F., Brown G., Galfre G., Milstein C., Williams A. F., Ziegler A. Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens-new tools for genetic analysis. Cell. 1978 May;14(1):9–20. doi: 10.1016/0092-8674(78)90296-9. [DOI] [PubMed] [Google Scholar]
  3. Bicknell D. C., Rowan A., Bodmer W. F. Beta 2-microglobulin gene mutations: a study of established colorectal cell lines and fresh tumors. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4751–4755. doi: 10.1073/pnas.91.11.4751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Chen H. L., Gabrilovich D., Virmani A., Ratnani I., Girgis K. R., Nadaf-Rahrov S., Fernandez-Viña M., Carbone D. P. Structural and functional analysis of beta2 microglobulin abnormalities in human lung and breast cancer. Int J Cancer. 1996 Sep 17;67(6):756–763. doi: 10.1002/(SICI)1097-0215(19960917)67:6<756::AID-IJC2>3.0.CO;2-Q. [DOI] [PubMed] [Google Scholar]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Cordon-Cardo C., Fuks Z., Drobnjak M., Moreno C., Eisenbach L., Feldman M. Expression of HLA-A,B,C antigens on primary and metastatic tumor cell populations of human carcinomas. Cancer Res. 1991 Dec 1;51(23 Pt 1):6372–6380. [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. D'Urso C. M., Wang Z. G., Cao Y., Tatake R., Zeff R. A., Ferrone S. Lack of HLA class I antigen expression by cultured melanoma cells FO-1 due to a defect in B2m gene expression. J Clin Invest. 1991 Jan;87(1):284–292. doi: 10.1172/JCI114984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ferrone S., Marincola F. M. Loss of HLA class I antigens by melanoma cells: molecular mechanisms, functional significance and clinical relevance. Immunol Today. 1995 Oct;16(10):487–494. doi: 10.1016/0167-5699(95)80033-6. [DOI] [PubMed] [Google Scholar]
  11. Fleischhauer K., Tanzarella S., Russo V., Sensi M. L., van der Bruggen P., Bordignon C., Traversari C. Functional heterogeneity of HLA-A*02 subtypes revealed by presentation of a MAGE-3-encoded peptide to cytotoxic T cell clones. J Immunol. 1997 Sep 1;159(5):2513–2521. [PubMed] [Google Scholar]
  12. Gattoni-Celli S., Kirsch K., Timpane R., Isselbacher K. J. Beta 2-microglobulin gene is mutated in a human colon cancer cell line (HCT) deficient in the expression of HLA class I antigens on the cell surface. Cancer Res. 1992 Mar 1;52(5):1201–1204. [PubMed] [Google Scholar]
  13. Goodfellow P. N., Jones E. A., Van Heyningen V., Solomon E., Bobrow M., Miggiano V., Bodmer W. F. The beta2-microglobulin gene is on chromosome 15 and not in the HL-A region. Nature. 1975 Mar 20;254(5497):267–269. doi: 10.1038/254267a0. [DOI] [PubMed] [Google Scholar]
  14. Gotch F., McMichael A., Smith G., Moss B. Identification of viral molecules recognized by influenza-specific human cytotoxic T lymphocytes. J Exp Med. 1987 Feb 1;165(2):408–416. doi: 10.1084/jem.165.2.408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Güssow D., Rein R., Ginjaar I., Hochstenbach F., Seemann G., Kottman A., Ploegh H. L. The human beta 2-microglobulin gene. Primary structure and definition of the transcriptional unit. J Immunol. 1987 Nov 1;139(9):3132–3138. [PubMed] [Google Scholar]
  16. Hicklin D. J., Kageshita T., Ferrone S. Development and characterization of rabbit antisera to human MHC-linked transporters associated with antigen processing. Tissue Antigens. 1996 Jul;48(1):38–46. doi: 10.1111/j.1399-0039.1996.tb02603.x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Lehmann F., Marchand M., Hainaut P., Pouillart P., Sastre X., Ikeda H., Boon T., Coulie P. G. Differences in the antigens recognized by cytolytic T cells on two successive metastases of a melanoma patient are consistent with immune selection. Eur J Immunol. 1995 Feb;25(2):340–347. doi: 10.1002/eji.1830250206. [DOI] [PubMed] [Google Scholar]
  19. Maio M., Altomonte M., Tatake R., Zeff R. A., Ferrone S. Reduction in susceptibility to natural killer cell-mediated lysis of human FO-1 melanoma cells after induction of HLA class I antigen expression by transfection with B2m gene. J Clin Invest. 1991 Jul;88(1):282–289. doi: 10.1172/JCI115289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marincola F. M., Hijazi Y. M., Fetsch P., Salgaller M. L., Rivoltini L., Cormier J., Simonis T. B., Duray P. H., Herlyn M., Kawakami Y. Analysis of expression of the melanoma-associated antigens MART-1 and gp100 in metastatic melanoma cell lines and in in situ lesions. J Immunother Emphasis Tumor Immunol. 1996 May;19(3):192–205. doi: 10.1097/00002371-199605000-00004. [DOI] [PubMed] [Google Scholar]
  21. Marincola F. M., Shamamian P., Alexander R. B., Gnarra J. R., Turetskaya R. L., Nedospasov S. A., Simonis T. B., Taubenberger J. K., Yannelli J., Mixon A. Loss of HLA haplotype and B locus down-regulation in melanoma cell lines. J Immunol. 1994 Aug 1;153(3):1225–1237. [PubMed] [Google Scholar]
  22. Matsui M., Temponi M., Ferrone S. Characterization of a monoclonal antibody-defined human melanoma-associated antigen susceptible to induction by immune interferon. J Immunol. 1987 Sep 15;139(6):2088–2095. [PubMed] [Google Scholar]
  23. Padgett R. A., Grabowski P. J., Konarska M. M., Seiler S., Sharp P. A. Splicing of messenger RNA precursors. Annu Rev Biochem. 1986;55:1119–1150. doi: 10.1146/annurev.bi.55.070186.005351. [DOI] [PubMed] [Google Scholar]
  24. Pandolfi F., Boyle L. A., Trentin L., Kurnick J. T., Isselbacher K. J., Gattoni-Celli S. Expression of HLA-A2 antigen in human melanoma cell lines and its role in T-cell recognition. Cancer Res. 1991 Jun 15;51(12):3164–3170. [PubMed] [Google Scholar]
  25. Pellegrino M. A., Ng A. K., Russo C., Ferrone S. Heterogeneous distribution of the determinants defined by monoclonal antibodies on HLA-A and B antigens bearing molecules. Transplantation. 1982 Jul;34(1):18–23. doi: 10.1097/00007890-198207000-00004. [DOI] [PubMed] [Google Scholar]
  26. Restifo N. P., Marincola F. M., Kawakami Y., Taubenberger J., Yannelli J. R., Rosenberg S. A. Loss of functional beta 2-microglobulin in metastatic melanomas from five patients receiving immunotherapy. J Natl Cancer Inst. 1996 Jan 17;88(2):100–108. doi: 10.1093/jnci/88.2.100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rivoltini L., Barracchini K. C., Viggiano V., Kawakami Y., Smith A., Mixon A., Restifo N. P., Topalian S. L., Simonis T. B., Rosenberg S. A. Quantitative correlation between HLA class I allele expression and recognition of melanoma cells by antigen-specific cytotoxic T lymphocytes. Cancer Res. 1995 Jul 15;55(14):3149–3157. [PMC free article] [PubMed] [Google Scholar]
  28. Stam N. J., Spits H., Ploegh H. L. Monoclonal antibodies raised against denatured HLA-B locus heavy chains permit biochemical characterization of certain HLA-C locus products. J Immunol. 1986 Oct 1;137(7):2299–2306. [PubMed] [Google Scholar]
  29. Versteeg R., Noordermeer I. A., Krüse-Wolters M., Ruiter D. J., Schrier P. I. c-myc down-regulates class I HLA expression in human melanomas. EMBO J. 1988 Apr;7(4):1023–1029. doi: 10.1002/j.1460-2075.1988.tb02909.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wang Z., Cao Y., Albino A. P., Zeff R. A., Houghton A., Ferrone S. Lack of HLA class I antigen expression by melanoma cells SK-MEL-33 caused by a reading frameshift in beta 2-microglobulin messenger RNA. J Clin Invest. 1993 Feb;91(2):684–692. doi: 10.1172/JCI116249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wang Z., Margulies L., Hicklin D. J., Ferrone S. Molecular and functional phenotypes of melanoma cells with abnormalities in HLA class I antigen expression. Tissue Antigens. 1996 May;47(5):382–390. doi: 10.1111/j.1399-0039.1996.tb02573.x. [DOI] [PubMed] [Google Scholar]
  32. Wölfel T., Klehmann E., Müller C., Schütt K. H., Meyer zum Büschenfelde K. H., Knuth A. Lysis of human melanoma cells by autologous cytolytic T cell clones. Identification of human histocompatibility leukocyte antigen A2 as a restriction element for three different antigens. J Exp Med. 1989 Sep 1;170(3):797–810. doi: 10.1084/jem.170.3.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yang S. Y., Morishima Y., Collins N. H., Alton T., Pollack M. S., Yunis E. J., Dupont B. Comparison of one-dimensional IEF patterns for serologically detectable HLA-A and B allotypes. Immunogenetics. 1984;19(3):217–231. doi: 10.1007/BF00364765. [DOI] [PubMed] [Google Scholar]
  34. van Duinen S. G., Ruiter D. J., Broecker E. B., van der Velde E. A., Sorg C., Welvaart K., Ferrone S. Level of HLA antigens in locoregional metastases and clinical course of the disease in patients with melanoma. Cancer Res. 1988 Feb 15;48(4):1019–1025. [PubMed] [Google Scholar]

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