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Functional CD8+ T cells infiltrate into nonsmall cell lung carcinoma

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Abstract

Infiltration of CD3+CD8+ cytotoxic T cells was analyzed by multiparameter confocal laser microscopy in a panel of 16 randomly selected stage I nonsmall cell lung carcinomas. T-cell infiltration was observed in the stroma (range 57-,093 T cells/mm2) but also in the tumor epithelium (range 21-92 T cells/mm2) and showed wide variation between individual tumors. Interestingly, a significantly higher percentage of CD3+CD8+ T cells was detected in the tumor epithelium compared to the stroma illustrating that cytotoxic T cells may preferentially migrate into tumor epithelium. Aberrant HLA class I antigen expression was observed in 69% of the nonsmall-cell lung carcinoma (NSCLC) tumors. One tumor of a squamous cell lung carcinoma patient with the highest number of tumor infiltrating CD3+ and CD3+CD8+ cells was studied in detail and the majority (90%) of these cells were shown to be functionally activated granzyme B-positive cytotoxic T cells. DNA oligotyping of a lung carcinoma cell line established from this tumor revealed loss of one HLA haplotype corresponding with a translocation involving chromosome 6, as observed by COBRA-FISH. HLA class I-restricted tumor specific T cells could be isolated from PBMC. One further characterized cytotoxic CD8+ T cell clone, that released TNF-α, IFN-γ, and granzyme B upon co-incubation with the autologous tumor cells, was shown to be restricted by the remaining HLA-A11 allele, which was also shown to be expressed in the tumor tissue. Our data indicate that, despite HLA-haplotype loss a vigorous antitumor immune response mediated by CD8+ T-cells can be present in NSCLC offering possibilities for specific immunotherapy.

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References

  1. Ambrosini G, Adida C, Altieri DC (1997) A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nat Med 3:917–921

    Article  CAS  PubMed  Google Scholar 

  2. Berrieman HK, Ashman JN, Cowen ME, Greenman J, Lind MJ, Cawkwell L (2004) Chromosomal analysis of non-small-cell lung cancer by multicolour fluorescent in situ hybridisation. Br J Cancer 90:900–905

    Article  CAS  PubMed  Google Scholar 

  3. Bladergroen BA, Meijer CJ, ten Berge RL, Hack CE, Muris JJ, Dukers DF, Chott A, Kazama Y, Oudejans JJ, van Berkum O, Kummer JA (2002) Expression of the granzyme B inhibitor, protease inhibitor 9, by tumor cells in patients with non-Hodgkin and Hodgkin lymphoma: a novel protective mechanism for tumor cells to circumvent the immune system? Blood 99:232–237

    Article  CAS  PubMed  Google Scholar 

  4. Boldrini L, Faviana P, Pistolesi F, Gisfredi S, Di Quirico D, Lucchi M, Mussi A, Angeletti CA, Baldinotti F, Fogli A, Simi P, Basolo F, Fontanini G (2001) Alterations of Fas (APO-1/CD 95) gene and its relationship with p53 in non small cell lung cancer. Oncogene 20:6632–6637

    Article  CAS  PubMed  Google Scholar 

  5. Carr NJ (2000) M30 expression demonstrates apoptotic cells, correlates with in situ end-labeling, and is associated with Ki-67 expression in large intestinal neoplasms. Arch Pathol Lab Med 124:1768–1772

    CAS  PubMed  Google Scholar 

  6. Deveraux QL, Takahashi R, Salvesen GS, Reed JC (1997) X-linked IAP is a direct inhibitor of cell-death proteases. Nature 388:300–304

    Article  CAS  PubMed  Google Scholar 

  7. Eerola AK, Soini Y, Paakko P (2000) A high number of tumor-infiltrating lymphocytes are associated with a small tumor size, low tumor stage, and a favorable prognosis in operated small cell lung carcinoma. Clin Cancer Res 6:1875–1881

    CAS  PubMed  Google Scholar 

  8. Fisher ER, Anderson S, Tan-Chiu E, Fisher B, Eaton L, Wolmark N (2001) Fifteen-year prognostic discriminants for invasive breast carcinoma: National Surgical Adjuvant Breast and Bowel Project Protocol-06. Cancer 91:1679–1687

    Article  CAS  PubMed  Google Scholar 

  9. Garrido F, Ruiz-Cabello F, Cabrera T, Perez-Villar JJ, Lopez-Botet M, Duggan-Keen M, Stern PL (1997) Implications for immunosurveillance of altered HLA class I phenotypes in human tumours. Immunol Today 18:89–95

    Article  CAS  PubMed  Google Scholar 

  10. Griffioen M, Borghi M, Schrier PI, Osanto S (2001) Detection and quantification of CD8(+) T cells specific for HLA-A*0201-binding melanoma and viral peptides by the IFN-gamma-ELISPOT assay. Int J Cancer 93:549–555

    Article  CAS  PubMed  Google Scholar 

  11. Hadden JW (1999) The immunology and immunotherapy of breast cancer: an update. Int J Immunopharmacol 21:79–101

    Article  CAS  PubMed  Google Scholar 

  12. Hilders CG, Munoz IM, Nooyen Y, Fleuren GJ (1995) Altered HLA expression by metastatic cervical carcinoma cells as a factor in impaired immune surveillance. Gynecol Oncol 57:366–375

    Article  CAS  PubMed  Google Scholar 

  13. Hiraki A, Ikeda K, Yoshino T, Kaneshige T, Kiura K, Kunisada T, Fujiwara K, Tanimoto M, Harada M (2001) Tumor-specific cytotoxic T lymphocyte responses against chondrosarcoma with HLA haplotype loss restricted by the remaining HLA class I allele. Biochem Biophys Res Commun 286:786–791

    Article  CAS  PubMed  Google Scholar 

  14. Ikeda H, Lethe B, Lehmann F, van Baren N, Baurain JF, de Smet C, Chambost H, Vitale M, Moretta A, Boon T, Coulie PG (1997) Characterization of an antigen that is recognized on a melanoma showing partial HLA loss by CTL expressing an NK inhibitory receptor. Immunity 6:199–208

    Article  CAS  PubMed  Google Scholar 

  15. Johnson SK, Kerr KM, Chapman AD, Kennedy MM, King G, Cockburn JS, Jeffrey RR (2000) Immune cell infiltrates and prognosis in primary carcinoma of the lung. Lung Cancer 27:27–35

    Article  CAS  PubMed  Google Scholar 

  16. Kataoka T, Shinohara N, Takayama H, Takaku K, Kondo S, Yonehara S, Nagai K (1996) Concanamycin A, a powerful tool for characterization and estimation of contribution of perforin- and Fas-based lytic pathways in cell-mediated cytotoxicity. J Immunol 156:3678–3686

    CAS  PubMed  Google Scholar 

  17. Kloor M, Becker C, Benner A, Woerner SM, Gebert J, Ferrone S, von Knebel DM (2005) Immunoselective pressure and human leukocyte antigen class I antigen machinery defects in microsatellite unstable colorectal cancers. Cancer Res 65:6418–6424

    Article  CAS  PubMed  Google Scholar 

  18. Kondratiev S, Sabo E, Yakirevich E, Lavie O, Resnick MB (2004) Intratumoral CD8+ T lymphocytes as a prognostic factor of survival in endometrial carcinoma. Clin Cancer Res 10:4450–4456

    Article  PubMed  Google Scholar 

  19. Lee SH, Shin MS, Park WS, Kim SY, Kim HS, Han JY, Park GS, Dong SM, Pi JH, Kim CS, Kim SH, Lee JY, Yoo NJ (1999) Alterations of Fas (Apo-1/CD95) gene in non-small cell lung cancer. Oncogene 18:3754–3760

    Article  CAS  PubMed  Google Scholar 

  20. Leers MP, Kolgen W, Bjorklund V, Bergman T, Tribbick G, Persson B, Bjorklund P, Ramaekers FC, Bjorklund B, Nap M, Jornvall H, Schutte B (1999) Immunocytochemical detection and mapping of a cytokeratin 18 neo-epitope exposed during early apoptosis. J Pathol 187:567–572

    Article  CAS  PubMed  Google Scholar 

  21. Menon AG, Janssen-van Rhijn CM, Morreau H, Putter H, Tollenaar RA, van de Velde CJ, Fleuren GJ, Kuppen PJ (2004) Immune system and prognosis in colorectal cancer: a detailed immunohistochemical analysis. Lab Invest 84:493–501

    Article  CAS  PubMed  Google Scholar 

  22. Menon AG, Morreau H, Tollenaar RA, Alphenaar E, Van Puijenbroek M, Putter H, Janssen-van Rhijn CM, van de Velde CJ, Fleuren GJ, Kuppen PJ (2002) Down-regulation of HLA-A expression correlates with a better prognosis in colorectal cancer patients. Lab Invest 82:1725–1733

    CAS  PubMed  Google Scholar 

  23. Muris JJ, Meijer CJ, Cillessen SA, Vos W, Kummer JA, Bladergroen BA, Bogman MJ, MacKenzie MA, Jiwa NM, Siegenbeek van Heukelom LH, Ossenkoppele GJ, Oudejans JJ (2004) Prognostic significance of activated cytotoxic T-lymphocytes in primary nodal diffuse large B-cell lymphomas. Leukemia 18:589–596

    Article  CAS  PubMed  Google Scholar 

  24. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139:271–279

    Article  CAS  PubMed  Google Scholar 

  25. Parham P, Barnstable CJ, Bodmer WF (1979) Use of a monoclonal antibody (W6/32) in structural studies of HLA-A,B,C, antigens. J Immunol 123:342–349

    CAS  PubMed  Google Scholar 

  26. Perosa F, Luccarelli G, Prete M, Favoino E, Ferrone S, Dammacco F (2003) Beta 2-microglobulin-free HLA class I heavy chain epitope mimicry by monoclonal antibody HC-10-specific peptide. J Immunol 171:1918–1926

    CAS  PubMed  Google Scholar 

  27. Piras F, Colombari R, Minerba L, Murtas D, Floris C, Maxia C, Corbu A, Perra MT, Sirigu P (2005) The predictive value of CD8, CD4, CD68, and human leukocyte antigen-D-related cells in the prognosis of cutaneous malignant melanoma with vertical growth phase. Cancer 104:1246–1254

    Article  CAS  PubMed  Google Scholar 

  28. Rajendra S, Ackroyd R, Karim N, Mohan C, Ho JJ, Kutty KM (2006) Loss of HLA class I and gain of class II expression are early events in carcinogenesis—clues from a study of Barrett’s oesophagus. J Clin Pathol (in press)

  29. Ramnath N, Tan D, Li Q, Hylander BL, Bogner P, Ryes L, Ferrone S (2006) Is downregulation of MHC class I antigen expression in human non-small cell lung cancer associated with prolonged survival? Cancer Immunol Immunother 55:891–899

    Article  CAS  PubMed  Google Scholar 

  30. Rebai N, Malissen B (1983) Structural and genetic analyses of HLA class I molecules using monoclonal xenoantibodies. Tissue Antigens 22:107–117

    Article  CAS  PubMed  Google Scholar 

  31. Redondo M, Concha A, Oldiviela R, Cueto A, Gonzalez A, Garrido F, Ruiz-Cabello F (1991) Expression of HLA class I and II antigens in bronchogenic carcinomas: its relationship to cellular DNA content and clinical–pathological parameters. Cancer Res 51:4948–4954

    CAS  PubMed  Google Scholar 

  32. Riemann D, Wenzel K, Schulz T, Hofmann S, Neef H, Lautenschlager C, Langner J (1997) Phenotypic analysis of T lymphocytes isolated from non-small-cell lung cancer. Int Arch Allergy Immunol 114:38–45

    Article  CAS  PubMed  Google Scholar 

  33. Roy N, Deveraux QL, Takahashi R, Salvesen GS, Reed JC (1997) The c-IAP-1 and c-IAP-2 proteins are direct inhibitors of specific caspases. EMBO J 16:6914–6925

    Article  CAS  PubMed  Google Scholar 

  34. Sandel MH, Speetjens FM, Menon AG, Albertsson PA, Basse PH, Hokland M, Nagelkerke JF, Tollenaar RA, van de Velde CJ, Kuppen PJ (2005) Natural killer cells infiltrating colorectal cancer and MHC class I expression. Mol Immunol 42:541–546

    Article  CAS  PubMed  Google Scholar 

  35. Schumacher K, Haensch W, Roefzaad C, Schlag PM (2001) Prognostic significance of activated CD8(+) T cell infiltrations within esophageal carcinomas. Cancer Res 61:3932–3936

    CAS  PubMed  Google Scholar 

  36. Seliger B (2005) Strategies of tumor immune evasion. BioDrugs 19:347–354

    Article  CAS  PubMed  Google Scholar 

  37. Sernee MF, Ploegh HL, Schust DJ (1998) Why certain antibodies cross-react with HLA-A and HLA-G: epitope mapping of two common MHC class I reagents. Mol Immunol 35:177–188

    Article  CAS  PubMed  Google Scholar 

  38. Stam NJ, Spits H, Ploegh HL (1986) Monoclonal antibodies raised against denatured HLA-B locus heavy chains permit biochemical characterization of certain HLA-C locus products. J Immunol 137:2299–2306

    CAS  PubMed  Google Scholar 

  39. Sy SM, Wong N, Lee TW, Tse G, Mok TS, Fan B, Pang E, Johnson PJ, Yim A (2004) Distinct patterns of genetic alterations in adenocarcinoma and squamous cell carcinoma of the lung. Eur J Cancer 40:1082–1094

    Article  CAS  PubMed  Google Scholar 

  40. Toomey D, Smyth G, Condron C, Kelly J, Byrne AM, Kay E, Conroy RM, Broe P, Bouchier-Hayes D (2003) Infiltrating immune cells, but not tumour cells, express FasL in non-small cell lung cancer: no association with prognosis identified in 3-year follow-up. Int J Cancer 103:408–412

    Article  CAS  PubMed  Google Scholar 

  41. Treilleux I, Blay JY, Bendriss-Vermare N, Ray-Coquard I, Bachelot T, Guastalla JP, Bremond A, Goddard S, Pin JJ, Barthelemy-Dubois C, Lebecque S (2004) Dendritic cell infiltration and prognosis of early stage breast cancer. Clin Cancer Res 10:7466–7474

    Article  CAS  PubMed  Google Scholar 

  42. van de Vijver MJ, Peterse JL, Mooi WJ, Wisman P, Lomans J, Dalesio O, Nusse R (1988) Neu-protein overexpression in breast cancer. Association with comedo-type ductal carcinoma in situ and limited prognostic value in stage II breast cancer. N Engl J Med 319:1239–1245

    Article  PubMed  Google Scholar 

  43. Verdegaal EM, Huinink DB, Hoogstraten C, Marijnissen AK, Gorsira MB, Claas FH, Osanto S (1999) Isolation of broadly reactive, tumor-specific, HLA Class-I restricted CTL from blood lymphocytes of a breast cancer patient. Hum Immunol 60:1195–1206

    Article  CAS  PubMed  Google Scholar 

  44. Verduyn W, Doxiadis II, Anholts J, Drabbels JJ, Naipal A, D’Amaro J, Persijn GG, Giphart MJ, Schreuder GM (1993) Biotinylated DRB sequence-specific oligonucleotides. Comparison to serologic HLA-DR typing of organ donors in eurotransplant. Hum Immunol 37:59–67

    Article  CAS  PubMed  Google Scholar 

  45. Wiegant J, Bezrookove V, Rosenberg C, Tanke HJ, Raap AK, Zhang H, Bittner M, Trent JM, Meltzer P (2000) Differentially painting human chromosome arms with combined binary ratio-labeling fluorescence in situ hybridization. Genome Res 10:861–865

    Article  CAS  PubMed  Google Scholar 

  46. Yang SY, Morishima Y, Collins NH, Alton T, Pollack MS, Yunis EJ, Dupont B (1984) Comparison of one-dimensional IEF patterns for serologically detectable HLA-A and B allotypes. Immunogenetics 19:217–231

    Article  CAS  PubMed  Google Scholar 

  47. Zhang L, Conejo-Garcia JR, Katsaros D, Gimotty PA, Massobrio M, Regnani G, Makrigiannakis A, Gray H, Schlienger K, Liebman MN, Rubin SC, Coukos G (2003) Intratumoral T cells, recurrence, and survival in epithelial ovarian cancer. N Engl J Med 348:203–213

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

Financial support for this study was granted by the Dutch Cancer Foundation, RUL19990-2027. We thank Prof. C.J M. Melief for critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Clinical Oncology, Leiden University Medical Center, P.O. Box 9600, 2300 RC, Leiden, The Netherlands

    Elizabeth M. E. Verdegaal, Conny Hoogstraten & Susanne Osanto

  2. Department of Surgery, Leiden University Medical Center, Leiden, The Netherlands

    Maro H. Sandel & Peter J. K. Kuppen

  3. Laboratory for Cytochemistry and Cytometry, Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands

    Antoinette A. T. P. Brink

  4. Department of Immunohaematology and Bloodbank, Leiden University Medical Center, Leiden, The Netherlands

    Frans H. J. Claas

  5. Department of Pathology, Diaconessenhuis Hospital, Leiden, The Netherlands

    Michael C. B. Gorsira & J. Frans Graadt van Roggen

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  1. Elizabeth M. E. Verdegaal
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Correspondence to Elizabeth M. E. Verdegaal.

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Verdegaal, E.M.E., Hoogstraten, C., Sandel, M.H. et al. Functional CD8+ T cells infiltrate into nonsmall cell lung carcinoma. Cancer Immunol Immunother 56, 587–600 (2007). https://doi.org/10.1007/s00262-006-0214-y

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  • DOI: https://doi.org/10.1007/s00262-006-0214-y

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