Skip to main content
PMC home page
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 2002 Feb;93(2):198–208. doi: 10.1111/j.1349-7006.2002.tb01259.x

Serine Proteinase Inhibitor 9 Can Be Recognized by Cytotoxic T Lymphocytes of Epithelial Cancer Patients

Koji Tanaka 1, Nanae Harashima 1, Fumihiko Niiya 2, Yoshiaki Miyagi 2, Naoya Hida 1, Mika Ochi 1, Nobue Imai 1, Mamoru Harada 1, Kyogo Itoh 1, Shigeki Shichijo 1,✉
PMCID: PMC5926951  PMID: 11856484

Abstract

Serine proteinase inhibitor 9 (PI–9) inhibits granzyme B‐mediated apoptosis and interleukin–lβ‐converting enzyme activity. In this study, we report that the PI–9 gene encodes antigenic epitopes recognized by the HLA‐A24–restricted and tumor‐reactive cytotoxic T lymphocytes (CTLs) of epithelial cancer patients. Screening of an autologous cDNA library using a CTL line recognizing HLA‐A24+ tumor cells resulted in the isolation of a cDNA, which had an identical coding region to the previously described PI–9 genes. PI–9 gene was expressed in approximately three‐fourths of epithelial cancer cell lines and all leukemic cell lines tested. It was also expressed in normal peripheral blood mononuclear cells (PBMCs), but not in a normal fibroblast cell line. CTL sublines contained T cells capable of recognizing the PI–9292–300 and PI–9348–356 peptides among 13 different peptides having the HLA‐A24 binding motifs. These two peptides were recognized by the CTL line in a dose‐dependent and HLA class‐I‐restricted manner, and also possessed the ability to induce HLA class I‐restricted and tumor‐reactive CTLs in PBMCs from HLA‐A24+ cancer patients. These results demonstrate that PI–9 is recognized by HLA class I‐restricted and tumor‐reactive CTLs of epithelial cancer patients.

Keywords: CTLs, PI–, 9, Serpin, Peptide

Full Text

The Full Text of this article is available as a PDF (400.4 KB).

REFERENCES

  • 1.van der Bruggen , P. , Traversari , C. , Chomez , P. , Lurqin , C. , Plaen , E. D. , van der Eynde , B. , Knuth , A. and Boon , T.A gene encoding an antigen recognized by cytotoxic T lymphocytes on a human melanoma . Science , 254 , 1643 – 1647 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 2.Kawakami , Y. , Eliyahu , S. , Sakaguchi , K. , Robbins , P. F. , Rivoltini , L. , Yannelli , J. B. , Appella , E. and 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. , 180 , 347 – 352 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Kawakami , Y. , Eliyahu , S. , Delgado , C. H. , Robbins , P. F. , Sakaguchi , K. , Appella , E. and Rosenberg , S. A.Identification of a human melanoma antigen recognized by tumor‐ infiltrating lymphocytes associated with in vivo tumor rejection . Proc. Natl. Acad. Set. USA , 91 , 6465 – 6462 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Brichard , V. , van Pel , A. , Wolfel , T. , de Plaen , E. , Lethe , B. , Coulie , P. and Boon , T.The tyrosinase gene codes for an antigen recognized by autologous cytolytic T lymphocytes on HLA‐A2 melanomas . J. Exp. Med. , 178 , 489 – 495 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Robbins , P. F. , EI‐Gamil , M. , Li , Y. F. , Kawakami , Y. , Loftus , D. , Appella , E. and Rosenberg , S. A.A mutated β‐catenin gene encodes a melanoma‐specific antigen recognized by tumor infiltrating lymphocytes . J. Exp. Med. , 183 , 1185 – 1192 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Guilloux , Y. , Lucas , S. , Brichard , V. G. , van Pel , A. , Viret , C. , Plaen , E. D. , Brasseur , F. , Lethe , B. , Joutereau , F. and Boon , T.A peptide recognized by human cytolytic T lymphocytes on HLA‐A2 melanoma is encoded by an intron sequence of the N‐acetylglucosaminyltranferase V gene . J. Exp. Med. , 183 , 1173 – 1183 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.van del Eynde , B. , Peeters , O. , De Backer , O. , Gauglar , B. , Lucas , S. and Boon , T.A new family of genes coding for an antigen recognized by autologous cytolytic T lymphocytes on a human melanoma . J. Exp. Med. , 182 , 689 – 697 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Lupetti , R. , Pisarra , P. , Verrecchia , A. , Farina , C. , Nicolini , G. , Anichini , A. , Bordignon , C. , Sensi , M. , Parmiani , G. and Traversari , C.Translation of a retained intron in tyrosinase‐related protein (TRP) 2 mRNA generates a new cytotoxic T lymphocytes (CTL)‐defined and shared human melanoma antigen not expressed in normal cells of the melanocytic lineage . J. Exp. Med. , 188 , 1005 – 1016 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Peoples , G. E. , Goedegebuure , P. S. , Smith , R. , Linehan , D. C. , Yoshino , I. and Eberlein , T. J.Breast and ovarian cancer‐specific cytotoxic T lymphocytes recognized the same HER–2/neu‐derived peptide . Proc. Natl. Acad. Sci. USA , 92 , 432 – 436 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Fisk , B. , Blevins , T. L. , Wharton , J. and loannides , C.Identification of an immunodominant peptide of HER–2/neu proto‐oncogene recognized by ovarian tumor‐specific cytotoxic T lymphocytes lines . J. Exp. Med. , 181 , 2109 – 2117 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Correale , P. , Walmsley , K. , Zaremba , S. , Zhu , M. , Schlom , J. and Tsang , K. Y.Generation of human cytolytic T lymphocyte lines directed against prostate‐specific antigen (PSA) employing a PSA oligoepitope peptide . J. Immunol. , 161 , 3186 – 3194 ( 1998. ). [PubMed] [Google Scholar]
  • 12.Correale , P. , Walmsley , K. , Nieroda , C. , Zaremba , S. , Zhu , M. , Schlom , J. and Tsang , K. Y.In vitro generation of human cytotoxic T lymphocytes specific for peptides derived from prostate‐specific antigen . J. Natl. Cancer Inst. , 89 , 293 – 300 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 13.Domenech , N. , Henderson , R. A. and Finn , O. J.Identification of an HLA‐A11–restricted epitope from the tandem repeat domain of the epithelial tumor antigen mucin . J. Immunol. , 155 , 4766 – 4774 ( 1995. ). [PubMed] [Google Scholar]
  • 14.Tsang , K. Y. , Zaremba , S. , Nieroda , C. A. , Zhu , M. Z. , Hamilton , J. M. and Schlom , J.Generation of human cytotoxic T cells specific for human carcinoembryonic antigen epitopes from patients immunized with recombinant vaccinia‐CEA vaccine . J. Natl. Cancer Inst. , 87 , 982 – 990 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 15.Shichijo , S. , Nakao , M. , Imai , Y. , Takasu , H. , Kawamoto , M. , Niiya , F. , Yang , D. , Tou , U. , Yamana , H. and Itoh , K.A gene encoding antigenic peptide of human squamous cell carcinoma recognized by cytotoxic T lymphocytes . J. Exp. Med. , 187 , 277 – 288 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Nakao , M. , Shichijo , S. , Imaizumi , T. , Inoue , Y. , Matsunaga , K. , Yamada , A. , Kikuchi , M. , Tsuda , N. , Ohta , K. , Takamori , S. , Yamana , H. , Fujita , H. and Itoh , K.Identification of a gene coding for a new squamous cell carcinoma antigen recognized by the CTL . J. Immunol. , 164 , 2565 – 2574 ( 2000. ). [DOI] [PubMed] [Google Scholar]
  • 17.Yang , D. , Nakao , M. , Shichijo , S. , Sasatomi , T. , Takasu , H. , Matsumoto , H. , Mori , K. , Hayashi , A. , Yamana , H. , Shirouzu , K. and Itoh , K.Identification of a gene coding for a protein possessing shared tumor epitopes capable of inducing HLA‐A24–restricted cytotoxic T lymphocytes in cancer patients . Cancer Res. , 59 , 4056 – 4063 ( 1999. ). [PubMed] [Google Scholar]
  • 18.Nishizaka , S. , Gomi , S. , Harada , K. , Oizumi , K. , Itoh , K. and Shichijo , S.A new tumor‐rejection antigen recognized by cytotoxic T lymphocytes infiltrating into a lung adeno‐carcinoma . Cancer Res. , 60 , 4830 – 4837 ( 2000. ). [PubMed] [Google Scholar]
  • 19.Kawano , K. , Gomi , S. , Tanaka , K. , Tsuda , N. , Kamura , T. , Itoh , K. and Yamada , A.Identification of a new endoplasmic reticulum‐resident protein recognized by HLA‐A24–restricted tumor‐infiltrating lymphocytes of lung cancer . Cancer Res. , 60 , 3550 – 3558 ( 2000. ). [PubMed] [Google Scholar]
  • 20.Gomi , S. , Nakao , M. , Niiya , F. , Imamura , Y. , Kawano , K. , Nishizaka , S. , Hayashi , A. , Sabao , Y. , Oizumi , K. and Itoh , K.A cyclophilin B gene encodes antigenic epitopes recognized by HLA‐A24–restricted and tumor‐specific CTLs . J. Immunol. , 163 , 4994 – 5004 ( 1999. ). [PubMed] [Google Scholar]
  • 21.Dahlen , J. R. , Foster , D. C. and Kisiel , W.Human proteinase inhibitor 9 (PI9) is a potent inhibitor of subtilisin A . Biochem. Biophys. Res. Commun. , 238 , 329 – 333 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 22.Sun , J. , Bird , C. H. , Sutton , V. , McDonald , L. , Coughlin , P. B. , Jong , T. A. D. , Trapan , J. A. and Bird , P. I.A cytosolic granzyme B inhibitor related to the viral apoptotic regulator cytokine response modifier is a present in cytotoxic lymphocytes . J. Biol. Chem. , 271 , 27802 – 27809 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 23.Sun , J. , Ooms , L. , Bird , C. H. , Sutton , V. R. , Trapani , J. A. and Bird , P. I.A new family of 10 murine ovalbumin serpins induces two homologs of proteinase inhibitor 8 and two homologs of the granzyme B inhibitor (Proteinase inhibitor 9) . J. Biol. Chem. , 272 , 15434 – 15441 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 24.Kanamori , H. , Krieg , S. , Mao , C. , Pippo , V. A. D. , Wang , S. , Zajchowski , D. A. and Shapiro , D. J.Proteinase inhibitor 9, an inhibitor of granzyme B‐mediated apoptosis, is a primary estrogen‐inducible gene in human liver cells . J. Biol. Chem. , 275 , 5867 – 5873 ( 2000. ). [DOI] [PubMed] [Google Scholar]
  • 25.Bird , C. H. , Sutton , V. R. , Sun , J. , Hirst , C. , Novak , A. , Kumar , S. , Trapani , J. A. and Bird , P. I.Selective regulation of apoptosis: the cytotoxic lymphocyte serpin proteinase inhibitor 9 protects against granzyme B‐mediated apoptosis without perturbing the Fas cell death pathway . Mol. Cell. Biol. , 18 , 6387 – 6398 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Kikuchi , M. , Nakao , M. , Inoue , Y. , Matsunaga , K. , Shichijo , S. , Yamana , H. and Itoh , K.Identification of a S ART–1–derived peptide capable of inducing HLA‐A24–restricted and tumor‐specific cytotoxic T lymphocytes . Int. J. Cancer , 81 , 459 – 466 ( 1999. ). [DOI] [PubMed] [Google Scholar]
  • 27.Ibe , M. , Moore , Y. L , Kaneko , Y. , Yokota , S. and Takiguchi , M.Role of strong anchor residues in the effective binding of 10–mer and 11–mer peptides to HLA‐A2402 molecules . Immunogenetics , 44 , 233 – 241 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 28.Parker , K. C. , Shields , M. , DiBrino , M. , Brooks , A. and Cologan , J. E.Peptide binding to MHC class I molecules: implications for antigenic peptide predictions . Immunol. Res. , 14 , 34 – 57 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 29.Lee , S. P. , Tiemey , R. J. , Thomas , W. A. , Brooks , J. M. and Rickinson , A. B.Conserved CTL epitopes within EBV latent membrane protein 2 . J. Immunol. , 158 , 3325 – 3334 ( 1997. ). [PubMed] [Google Scholar]
  • 30.Schumacher , T. M. , Heemels , M. T. , Neefjes , J. J. , Kast , W.M. , Melief , C. J. M. and Ploegh , H. L.Direct binding of peptide to empty MHC class I molecules on intact cells and in vitro . Cell , 62 , 563 – 567 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 31.Young , J. L. , Sukhova , G. K. , Foster , D. , Kisiel , W. , Libby , P. and Schonbeck , U.The serpin proteinase inhibitor 9 is an endogenous inhibitor of interleukin–lβ‐converting enzyme (Caspase–1) activity is human vascular smooth muscle cells . J. Exp. Med. , 191 , 1535 – 1544 ( 2000. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Annand , R. R. , Dahlen , J. R. , Sprecher , C. A. , De Dreu , P. , Foster , D. C. , Mankovich , J. A. , Talanian , R. V. , Kisiel , W. and Giegel , D. A.Caspase–1 (interleukin–lβ‐converting enzyme) is inhibited by the human serpin analogue proteinase inhibitor 9 . Biochem. J. , 342 , 655 – 665 ( 1999. ). [PMC free article] [PubMed] [Google Scholar]
  • 33.Sprecher , C. A. , Morgenstern , K. A. , Mathewes , S. , Dahlen , J. R. , Schrader , S. K. , Foster , D. C. and Kisiel , W.Molecular cloning expression, and partial characterization of two novel members of the ovalbumin family of serine proteinase inhibitors . J. Biol. Chem. , 270 , 29854 – 29861 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 34.Bladergroen , B. A. , Strik , M. C. M. , Bovenschen , N. , van Berkum , O. , Scheffer , G. L. , Meijer , C. J. L. M. , Hack , C. E. and Kummer , J. A.The granzyme B inhibitor, protease inhibitor 9, is mainly expressed by dendritic cells and at immune‐privileged sites . J. Immunol. , 166 , 3218 – 3225 ( 2001. ). [DOI] [PubMed] [Google Scholar]
  • 35.Harashima , N. , Tanaka , K. , Sasatomi , T. , Shimizu , K. , Miyagi , Y. , Yamada , A. , Yamana , H. , Itoh , K. and Shichijo , S.Recognition of the Lck tyrosinase kinase as a tumor antigen by cytotoxic T lymphocytes of cancer patients with distant metastases . Eur. J. Immunol. , 31 , 323 – 332 ( 2001. ). [DOI] [PubMed] [Google Scholar]
  • 36.Schmitz , M. , Diestelkoetter , P. , Weigle , B. , Schmach‐tenberg , F. , Stevanovic , S. , Ockert , D. , Rammen‐see , H.‐G. and Rieber , E. P.Generation of survivin‐specific CD8+ T effector cells by dendritic cells pulsed with protein or selected peptides . Cancer Res. , 60 , 4845 – 4849 ( 2000. ). [PubMed] [Google Scholar]
  • 37.Andersen , M. H. , Pedersen , L. O. , Becker , J. C. and Straten , P. T.Identification of a cytotoxic T lymphocyte response to the apoptosis inhibitor protein survivin in cancer patients . Cancer Res. , 61 , 869 – 872 ( 2001. ). [PubMed] [Google Scholar]

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

RESOURCES