Skip to main content
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1995 May;86(5):477–483. doi: 10.1111/j.1349-7006.1995.tb03081.x

Functional and T Cell Receptor Gene Usage Analysis of Cytotoxic T Lymphocytes in Fresh Tumor‐infiltrating Lymphocytes from Human Head and Neck Cancer

Kazuaki Chikamatsu 1,, Masao Eura 1, Koji Nakano 1, Keisuke Masuyama 1, Takeru Ishikawa 1
PMCID: PMC5920848  PMID: 7790320

Abstract

Twenty‐one cytotoxic T lymphocyte (CTL) clones or lines that killed autologous tumor cells, but not allogeneic tumor, K562, or Daudi cells, were established from fresh tumor‐infiltrating lymphocytes of two individuals (HP‐1 and HP‐2) with head and neck cancer by limiting dilution in the presence of recombinant interleukin‐2. Sixteen (76%) of these 21 clones or lines comprised CD4+ CTLs and the other five comprised CD8+ CTLs. These observations suggest that autologous tumor cell‐specific CD4+ CD8 and CD4 CD8+ CTLs are present in vivo at the tumor site in head and neck cancer. Analysis of T cell receptor (TCR) gene arrangements in 20 of the 21 CTL isolates with reverse transcriptase and the polymerase chain reaction revealed that five of 12 and five of eight isolates from HP‐1 and HP‐2, respectively, were clones, the other isolates being lines comprised of two or more clones. Each CTL clone showed a different combination of Vα and Vβ gene expression, suggesting that more than five different tumor‐associated antigens may be expressed on head and neck cancer cells. In spite of the diversity of TCR αβ combinations, TCR Vα1, Vα3, Vα8, Vα10, Vβ8, and Vβ9, and Vβ17 were also frequently expressed in both patients. These data suggest that specific CTLs proliferate oligoclonally and contribute to the specific immune response against head and neck cancer in vivo.

Keywords: T cell receptor, ytotoxic T lymphocyte, quamous cell carcinoma, umor‐associated antigen, Gene usage

Full Text

The Full Text of this article is available as a PDF (409.2 KB).

REFERENCES

  • 1).Itoh , K. , Platsoucas , C. D. and Balch , C. M.Autologous tumor‐specific cytotoxic T lymphocytes in the infiltrate of human metastatic melanomas . J. Exp. Med. , 168 , 1419 – 1441 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2).Gervois , N. , Heuze , F. , Diez , E. and Jotereau , F.Selective expansion of a specific anti‐tumor CD8+ cytotoxic T lymphocyte clone in the bulk culture of tumor‐infiltrating lymphocytes from a melanoma patient: cytotoxic activity and T cell receptor gene rearrangements . Eur. J. Immuno. , 20 , 825 – 831 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 3).Ioannides , C. G. , Freedman , R. S. , Platsoucas , C. D. , Rashed , S. and Kim , Y. P.Cytotoxic T cell clones isolated from ovarian tumor‐infiltrating lymphocytes recognize multiple antigenic epitopes on autologous tumor cells . J. Immunol , 146 , 1700 – 1707 ( 1991. ). [PubMed] [Google Scholar]
  • 4).Ikeda , H. , Sato , N. , Matsuura , A. and Kikuchi , K.Analysis of T‐cell receptor V region gene usage of cytotoxic T‐lymphocytes and tumor‐infiltrating lymphocytes derived from human autologous gastric signet ring cell carcinomas . Cancer Res. , 53 , 3078 – 3084 ( 1993. ). [PubMed] [Google Scholar]
  • 5).Shimizu , Y. , Weidmann , E. , Iwatsuki , S. , Herberman , R. B. and Whiteside , T. L.Characterization of human autotumor‐reactive T‐cell clones obtained from tumor‐infiltrating lymphocytes in liver metastasis of gastric carcinoma . Cancer Res , 51 , 6153 – 6162 ( 1991. ). [PubMed] [Google Scholar]
  • 6).Heo , D. S. , Whiteside , T. L. , Johnson , J. T. , Chen , K. , Barnes , E. L. and Herberman , R. B.Long‐term interleu‐kin 2‐dependent growth and cytotoxic activity of tumor‐infiltrating lymphocytes from human squamous cell carcinomas of the head and neck . Cancer Res. , 47 , 6353 – 6362 ( 1987. ). [PubMed] [Google Scholar]
  • 7).Letessier , E. M. , Heo , D. S. , Okarma , T. , Johnson , J. T. , Herberman , R. B. and Whiteside , T. L.Enrichment in tumor‐reactive CD8+ T‐lymphocytes by positive selection from the blood and lymph nodes of patients with head and neck cancer . Cancer Res. , 51 , 3891 – 3899 ( 1991. ). [PubMed] [Google Scholar]
  • 8).Yasumura , S. , Hirabayashi , H. , Schwartz , D. R. , Toso , J. F. , Johnson , J. T. , Herberman , R. B. and Whiteside , T. L.Human cytotoxic T‐cell lines with restricted specificity for squamous cell carcinoma of the head and neck . Cancer Res. , 53 , 1461 ( 1993. ). [PubMed] [Google Scholar]
  • 9).van der Bruggen , P. , Traversari , C. , Chomez , P. , Lurquin , C. , De Plaen , E. , Van den Eynde , B. , Knuth , A. and Boon , T.A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma . Science , 254 , 1643 – 1647 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 10).Traversari , C. , van der Bruggen , P. , Luescher , I. F. , Lurquin , C. , Chomez , P. , Van Pel , A. , De Plaen , E. , Amar‐Costesec , A. and Boon , T.A nonapeptide encoded by human gene MAGE‐1 is recognized on HLA‐A1 by cytolytic T lymphocytes directed against tumor antigen MZ2‐E . J. Exp. Med. , 176 , 1453 – 1457 ( 1992. ). [DOI] [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. , Lethe , B. , Brasseur , F. and Boon , T.Human gene MAGE‐3 codes for an antigen recognized on a melanoma by autologous cytolytic T lymphocytes . J. Exp. Med. , 179 , 921 – 930 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12).Brasseur , F. , Marchand , M. , Vanwijck , R. , Herin , M. , Lethe , B. , Chomez , P. and Boon , T.Human gene MAGE‐1, which codes for a tumor‐rejection antigen, is expressed by some breast tumors . Int. J. Cancer , 52 , 839 – 841 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 13).Rimoldi , D. , Romero , P. and 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 , 54 , 527 – 528 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 14).Weynants , P. , Lethe , B. , Brasseur , F. , Marchand , M. and Boon , T.Expression of MAGE genes by non‐small‐cell lung carcinomas . Int. J. Cancer , 56 , 826 – 829 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 15).Nitta , T. , Oksenberg , J. R. , Rao , N. A. and Steinman , L.Predominant expression of T cell receptor Vα7 in tumor‐infiltrating lymphocytes of uveal melanoma . Science , 249 , 672 – 674 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 16).Ferradini , L. , Roman‐Roman , S. , Azocar , J. , Avril , M. F. , Viel , S. , Triebel , F. and Hercend , T.Analysis of T‐cell receptor αβ variability in lymphocytes infiltrating a melanoma metastasis . Cancer Res , 52 , 4649 – 4654 ( 1992. ). [PubMed] [Google Scholar]
  • 17).Chikamatsu , K. , Eura , M. , Nakano , K. , Kanzaki , Y. , Matsuoka , H. , Masuyama , K. and Ishikawa , T.Analysis of T cell receptor variability in fresh tumor‐infiltrating lymphocytes from human head and neck cancer . Jpn. J. Cancer Res , 85 , 626 – 632 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18).Chikamatsu , K. , Eura , M. , Matsuoka , H. , Murakami , H. , Fukiage , T. and Ishikawa , T.The role of major histo‐compatibility complex expression on head and neck cancer cells in the induction of autologous cytotoxic T lymphocytes . Cancer Immunol. Immunother. , 38 , 358 – 364 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19).Ichino , Y. and Ishikawa , T.Effects of cryopreservation on human lymphocyte functions: comparison of programmed freezing method by a direct control system with a mechanical freezing method . J. Immunol. Methods , 77 , 283 – 290 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 20).Moretta , A. , Pantaleo , G. , Moretta , L. , Cerottini , J. C. and Mingari , M. C.Direct demonstration of the clonogenic potential of every human peripheral blood T cell. Clonal analysis of HLA‐DR expression and cytolytic activity . Exp. Med. , 157 , 743 – 754 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21).Schendel , D. J. , Gransbacher , B. , Oberneder , R. , Kriegmair , M. , Hofstetter , A. , Riethmuller , G. and Segurado , O. G.Tumor‐specific lysis of human renal cell carcinomas by tumor‐infiltrating lymphocytes. I. HLA‐A2‐restricted recognition of autologous and allogeneic tumor lines . J. Immunol. , 151 , 4209 – 4220 ( 1993. ). [PubMed] [Google Scholar]
  • 22).Herin , M. , Lemoine , C. , Weynants , P. , Vessiere , F. , Van Pel , A. , Knuth , A. , Devos , R. and Boon , T.Production of stable cytolytic T‐cell clones directed against autologous human melanoma . Int. J. Cancer , 39 , 390 – 396 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 23).Wolfel , T. , Klehmann , E. , Muller , C. , Schutt , K. H. , Meyer Zum Buschenfelde , K. H. and 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. , 170 , 797 – 810 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24).Wolfel , T. , Hauer , M. , Klehmann , E. , Brichard , V. , Ackermann , B. , Knuth , A. , Boon , T. and Meyer Zum Buschenfelde , K. H.Analysis of antigens recognized on human melanoma cells by A2‐restricted cytolytic T lymphocytes (CTL) . Int. J. Cancer , 55 , 237 – 244 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 25).Anichini , A. , Fossati , G. and Parmiani , G.Clonal analysis of the cytolytic T‐cell response to human tumors . Immunol. Today , 8 , 385 – 389 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 26).Peoples , G. E. , Davey , M. P. , Goedegebuure , P. S. , Schoof , D. D. and Eberlein , T. J.T cell receptor Vβ2 and Vβ6 mediate tumor‐specific cytotoxicity by tumor‐infiltrating lymphocytes in ovarian cancer . J. Immunol. , 151 , 5472 – 5480 ( 1993. ). [PubMed] [Google Scholar]
  • 27).Sensi , M. , Salvi , S. , Castelli , C. , Maccalli , C. , Mazzocchi , A. , Mortarini , R. , Nicolini , G. , Herlyn , M. , Parmiani , G. and Anichini , A.T cell receptor (TCR) structure of autologous melanoma‐reactive cytotoxic T lymphocyte (CTL) clones: tumor‐infiltrating lymphocytes overexpress in vivo the TCR β chain sequence used by an HLA‐A2‐restricted and melanocyte‐lineage‐specific CTL clone . J. Exp. Med. , 178 , 1231 – 1246 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28).Brichard , V. , Van Pel , A. , Wolfel , T. , Wolfel , C. , De Plaen , E. , Lethe , B. , Coulie , P. and Boon , T.The tyrosi‐nase gene codes for an antigen recognized by autologous cytolytic T lymphocytes on HLA‐A2 melanomas . J. Exp. Med. , 178 , 489 – 495 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES