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Analysis of T cell receptor β and γ genes from peripheral blood, regional lymph node and tumor-infiltrating lymphocyte clones from melanoma patients

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Summary

A total of 199 T cell clones from two melanoma patients were derived from progenitor T cells from recurrent melanoma, regional lymph nodes (either involved or uninvolved with malignancy) and peripheral blood by inoculating single cells directly into the wells of microtiter plates before in vitro expansion. The surface marker phenotype of most clones was CD4+CD8−, although some were CD4−CD8+. Genomic DNA prepared from all clones was analyzed by Southern blot hybridization using T cell receptor (TCR) β and γ gene probes, seeking clones with identical TCR gene rearrangement patterns as direct evidence for in vivo progenitor T cell clonal amplification. ProbingHindIII-digested DNA with TCRβ and TCRγ probes revealed several clones with identical TCR gene rearrangement patterns. These clones had subsequent probing ofBamHI-digested DNA with TCRβ and TCRγ probes, which showed all but 2 clones to have distinct rearrangement patterns. These analyses provide clear molecular evidence for in vivo polyclonal CD4+ T cell populations in each of several separate immune compartments in these patients.

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References

  1. Acha-Orbea H, Mitchell D, Timmermann L, Wraith D, Tausch G, Waldor M, Zamvil S, McDevitt H, Steinman L (1988) Limited heterogeneity of T-cell receptors from lymphocytes mediating autoimmune encephalomyelitis allows specific immune intervention. Cell 54: 263–273

    Google Scholar 

  2. Albertini RJ (1985) Somatic gene mutations in vivo as indicated by the 6-thioguanine-resistant T-lymphocytes in human blood. Mutat Res 150: 411–422

    Google Scholar 

  3. Anichini A, Fossati G, Parmianio G (1987) Clonal analysis of the cytolytic T-cell response to human tumors. Immunol Today 8: 385–389

    Google Scholar 

  4. Eynde VC, Hainaut P, Herin M, Knuth A, Lemoine C, Weynants P, Bruggen PVD, Fauchet R, Boon T (1989) Presence on a human melanoma of multiple antigens recognized by autologous CTL. Int J Cancer 44: 634–640

    Google Scholar 

  5. Fishleder AJ, Finke JH, Tubbs R, Bukowski RM (1990) Induction by interleukin-2 of oligoclonal expansion of cultured tumor-infiltrating lymphocytes. J Natl Cancer Inst 82: 124–128

    Google Scholar 

  6. Fleischer B, Bogdahn U (1983) Growth of antigen specific, HLA restricted T-lymphocyte clones from cerebrospinal fluid. Clin Exp Immunol 52: 38–44

    Google Scholar 

  7. Fleischer B, Marquardt P, Poser S, Kreth H (1984) Phenotypic markers and functional characteristics of T-lymphocyte clones from cerebrospinal fluid in multiple sclerosis. J Neuroimmunol 7: 151–162

    Google Scholar 

  8. Fliedner V, Qiao L, Whiteside T, Leyvraz S, Barras C, Miescher S (1987) Clonogenic and functional potential of T-lymphocytes infiltrating human solid tumors. In: Truitt R, Gale R, Bortin M (eds) Cellular immunotherapy of cancer. Liss, New York, pp 223–232

    Google Scholar 

  9. Galili U, Vanky F, Rodriguez L, Klein E (1979) Activated T-lymphocytes within human solid tumors. Cancer Immunol Immunother 6: 129–133

    Google Scholar 

  10. Hamaoka T, Fujiwara H (1987) Phenotypically and functionally distinct T-cell subsets in antitumor responses. Immunol Today 8: 267–269

    Google Scholar 

  11. Hersey P, Bolhuis R (1987) ‘Nonspecific’ MHC-unrestricted killer cells and their receptors. Immunol Today 8: 233–239

    Google Scholar 

  12. Itoh K, Tilden A, Balch C (1986) Interleukin-2 activation of cytotoxic T-lymphocytes infiltrating into human metastatic melanomas. Cancer Res 46: 3011–3017

    Google Scholar 

  13. Itoh K, Platsoucas C, Balch C (1988) Autologous tumor-specific cytotoxic T-lymphocytes in the infiltrate of human metastatic melanoma. J Exp Med 168: 1419–1441

    Google Scholar 

  14. Knuth A, Wolfel T, Klehmann E, Boon T, Buschenfelde K (1989) Cytolytic T-cell clones against an autologous melanoma: specificity study and definition of three antigens by immunoselection. Proc Natl Acad Sci USA 86: 2804–2808

    Google Scholar 

  15. Lanier L, Phillips J (1986) Evidence for three types of human cytotoxic lymphocytes. Immunol Today 7: 132–134

    Google Scholar 

  16. Lee SJ, Benjamin D, Weiner H, Seidman J, Duby A, Ang S, Hafler D (1988) Clonality and T-cell receptor γ chains in immune compartments: examination of cerebrospinal fluid derived T-cell clones. In: Davis M, Kappler J (eds) The T-cell receptor. Liss, New York, pp 87–97

    Google Scholar 

  17. Lefranc M, Forster A, Rabbitts T (1986) Rearrangement of two distinct T-cell γ-chain variable-region genes in human DNA. Nature 319: 420–422

    Google Scholar 

  18. Marrack P, Kappler J (1986) The antigen-specific, major histocompatibilitycomplex-restrictedreceptoron T-cells.AdvImmunol38: 1–30

    Google Scholar 

  19. Marrack P, Kappler J (1987) The T-cell receptor. Science 238: 1073–1078

    Google Scholar 

  20. Miescher S, Stoeck M, Qigo L, Barras C, Barrelet L, Fliedner V (1988) Preferential clonogenic deficit of CD8-positive T-lymphocytes infiltrating human solid tumors. Cancer Res 48: 6992–6998

    Google Scholar 

  21. Modlin R, Brenner M, Krangel M, Duby A, Bloom B (1987) T-cell receptors of human suppressor cells. Nature 329: 541–545

    Google Scholar 

  22. Moller D, Komishi K, Kirby M, Balbi B, Crystal R (1988) Bias toward use of a specific T-cell receptor β-chain variable region in a subgroup of individuals with sarcoidosis. J Clin Invest 82: 1183–1191

    Google Scholar 

  23. Mukherji B, Guha A, Chakraborty N, Sivanandham M, Nashed A, Sporn J, Ergin M (1989) Clonal analysis of cytotoxic and regulatory T-cell responses against human melanoma. J Exp Med 169: 1961–1979

    Google Scholar 

  24. Nicklas JA, Hunter T, Sullivan L, Berman J, O'Neill J, Albertini R (1987) Molecular analysis of in vivo hprt mutations in human T-lymphocytes: I. Studies of low frequency ‘spontaneous’ mutants by Southern blots. Mutagenesis 2: 341–347

    Google Scholar 

  25. Nicklas JA, Hunter T, O'Neill J, Albertini R (1989) Molecular analyses of in vivo hprt mutations in human T-lymphocytes: III. Longitudinal study of hprt gene structural alterations and T-cell clonal origins. Mutation Res 215: 147–160

    Google Scholar 

  26. Nitta T, Oksenberg JR, Rao NA, Steinman L (1990) Predominant expression of T-cell receptor Vα7 in tumor-infiltrating lymphocytes of uveal melanoma. Science 249: 672–674

    Google Scholar 

  27. O'Neill JP, McGinniss M, Berman J, Sullivan L, Nicklas J, Albertini R (1987) Refinement of a T lymphocyte cloning assay to quantify the in vivo thioguanine-resistant mutant frequency in humans. Mutagenesis 2: 87–94

    Google Scholar 

  28. Padula SJ, Sjroi D, Lingenheld E, Love J Jr, Chou C, Clark R (1988) T-Cell receptor beta chain gene rearrangement shared by murine T-cell lines derived from a site of autoimmune inflammation. J Clin Invest 81: 1810–1818

    Google Scholar 

  29. Pardoll D, Kruisbeek A, Fowlkes B, Coligan J, Schwartz R (1987) The unfolding story of T-cell receptor γ. FASEB J 1: 103–109

    Google Scholar 

  30. Poljak R (1987) Hypothesis: a molecular model for the MHC-restricted recognition of antigens by the T-cell receptor. Ann Inst Pasteur Immunol 138: 175–180

    Google Scholar 

  31. Reinherz E (1987) T-Cell receptors: who needs more? Nature 660–663

  32. Rosenberg SA, Packard B, Aebersold P, Solomon D, Topalian S, Toy S, Simon P, Lotze M, Yang J, Seipp C, Simpson C, Carter C, Bock S, Schwartzentruber D, Wei J, White D (1988) Use of tumorinfiltrating lymphocytes and interleukin-2 in the immunotherapy of patients with metastatic melanoma. N Engl J Med 319: 1676–1680

    Google Scholar 

  33. Royer D, Reinherz E (1987) T-lymphocytes: ontogeny, function, and relevance to clinical disorders. N Engl J Med 317: 1136–1142

    Google Scholar 

  34. Sondel PM, Hank J, Kohler P, Chen B, Sosman J (1989) Lymphokine activated killing as treatment for human cancer: clinical extrapolations from laboratory studies with interleukin-2 expanded leukocytes. In: Mitchell M (ed) Immunity to cancer; II. pp. 151–160, Liss, New York, pp 151–161

    Google Scholar 

  35. Stamenkovic I, Stegagno M, Wright K, Krane S, Amento E, Colvin R, Duquesnoy R, Kurnick J (1988) Clonal dominance among T-lymphocyte infiltrates in arthritis. Proc Natl Acad Sci USA 85: 1179–1183

    Google Scholar 

  36. Thompson D, Sutherland M, Scanzano R, Shenouda G (1987) Recognition of a purifiedM r-40,000 organ specific immunogen of human lung cancer by class II-restricted T-cells. JNCI 79: 653–661

    Google Scholar 

  37. Yanagi Y, Yoshikai Y, Leggett K, Clark S, Alekgander I, Mak T (1984) A human T-cell-specific cDNA clone encodes a protein having extensive homology to immunoglobulin chains. Nature 308: 145–149

    Google Scholar 

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Author notes

  1. Mark R. Albertini

    Present address: K4/450, Clinical Science Center, 600 Highland Avenue, 53 792, Madison, WI, USA

Authors and Affiliations

  1. Department of Human Oncology, University of Wisconsin, 53 792, Madison, WI, USA

    Mark R. Albertini, Steven S. Clark, Jacquelyn A. Hank & Paul M. Sondel

  2. Department of Pediatrics, University of Wisconsin, 53 792, Madison, WI, USA

    Paul M. Sondel

  3. Department of Genetics, University of Wisconsin, 53 792, Madison, WI, USA

    Paul M. Sondel

  4. Genetics Laboratory, University of Vermont, 05 401, Burlington, Vermont, USA

    Janice A. Nicklas, Bettejayne F. Chastenay, Timothy C. Hunter & Richard J. Albertini

Authors
  1. Mark R. Albertini
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  2. Janice A. Nicklas
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  3. Bettejayne F. Chastenay
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  4. Timothy C. Hunter
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  5. Richard J. Albertini
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  6. Steven S. Clark
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  7. Jacquelyn A. Hank
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  8. Paul M. Sondel
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Additional information

This investigation was supported by National Institutes of Health, National Research Service Award CA-08 397 from the National Cancer Institute as well as NIH CA-32 685, CA-30 688, DOE FG028 760 502 and American Cancer Society Grant ACS CH-237

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Albertini, M.R., Nicklas, J.A., Chastenay, B.F. et al. Analysis of T cell receptor β and γ genes from peripheral blood, regional lymph node and tumor-infiltrating lymphocyte clones from melanoma patients. Cancer Immunol Immunother 32, 325–330 (1991). https://doi.org/10.1007/BF01789051

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  • DOI: https://doi.org/10.1007/BF01789051

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