Abstract
Purpose
The efficient identification of peptide antigens recognized by ovarian cancer-specific cytotoxic T lymphocytes (CTL) requires the use of well-characterized ovarian cancer cell lines. To develop such a panel of cell lines, 11 ovarian cancer cell lines were characterized for the expression of class I and class II major histocompatibility complex (MHC)-encoded molecules, 15 tumor antigens, and immunosuppressive cytokines [transforming growth factor β (TGF-β) and IL-10].
Methods
Class I MHC gene expression was determined by polymerase chain reaction (PCR), and class I and class II MHC protein expression was determined by flow cytometry. Tumor antigen expression was determined by a combination of polymerase chain reaction (PCR) and flow cytometry. Cytokine expression was determined by ELISA.
Results
Each of the ovarian cancer cell lines expresses cytokeratins, although each cell line does not express the same cytokeratins. One of the lines expresses CD90, which is associated with a fibroblast lineage. Each of the cell lines expresses low to moderate amounts of class I MHC molecules, and several of them express low to moderate amounts of class II MHC molecules. Using a combination of PCR and flow cytometry, it was determined that each cell line expressed between six and thirteen of fifteen antigens tested. Little to no TGF-β3 was produced by any of the cell lines, TGF-β1 was produced by three of the cell lines, TGF-β2 was produced by all of the cell lines, with four of the cell lines producing large amounts of the latent form of the molecule, and IL-10 was produced by one of the cell lines.
Conclusions
Each of the 11 ovarian cancer lines is characterized by a unique expression pattern of epithelial/fibroblast markers, MHC molecules, tumor antigens, and immunosuppressive cytokines. Knowledge of these unique expression patterns will increase the usefulness of these cell lines in identifying the antigens recognized by ovarian cancer-specific CTL.
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Abbreviations
- B-LCL:
-
B-lymphoblastoid cell line
- CEA:
-
Carcinoembryonic antigen
- CTL:
-
Cytotoxic T lymphocyte
- Ck:
-
Cytokeratin
- FBP:
-
Folate binding protein
- FBS:
-
Fetal bovine serum
- GAPDH:
-
Glyceraldehyde-3-phosphate dehydrogenase
- mAb:
-
Monoclonal antibody
- MHC:
-
Major histocompatibility complex
- PCR:
-
Polymerase chain reaction
- TGF-β:
-
Transforming growth factor β
References
Ahmadzadeh M, Rosenberg SA (2005) TGF-β1 attenuates the acquisition and expression of effector function by tumor antigen-specific human memory CD8 T cells. J Immunol 174:5215–5223
Akdis CA, Blaser K, Akdis CA, Blaser K (2001) Mechanisms of interleukin-10-mediated immune suppression. Immunology 103:131–136
Angus B, Purvis J, Stock D, Westley BR, Samson ACR, Routledge EG, Carpenter FH, Horne CHW (1987) NCL-5D3: a new monoclonal antibody recognizing low molecular weight cytokeratins effective for immunohistochemistry using fixed paraffin-embedded tissue. J Pathol 153:377–384
Babcock B, Anderson BW, Papayannopoulos I, Castilleja A, Murray JL, Stifani S, Kudelka AP, Wharton JT, Ioannides CG (1998) Ovarian and breast cytotoxic T lymphocytes can recognize peptides from the amino enhancer of split protein of the notch complex. Mol Immunol 35:1121–1133
Bartlett JM, Langdon SP, Scott WN, Love SB, Miller EP, Katsaros D, Smyth JF, Miller WR (1997) Transforming growth factor-β isoform expression in human ovarian tumours. Eur J Cancer 33:2397–2403
Berger AE, Davis JE, Cresswell P (1982) Monoclonal antibody to HLA-A3. Hybridoma 1:87–90
Berger S, Siegert A, Denkert C, Kobel M, Hauptmann S (2001) Interleukin-10 in serous ovarian carcinoma cell lines. Cancer Immunol Immunother 50:328–333
Brasseur F, Marchand M, Vanwijck R, Herin M, Lethe B, Chomez P, Boon T (1992) Human gene MAGE-1, which codes for a tumor-rejection antigen, is expressed by some breast tumors. Int J Cancer 52:839–841
Brunetti M, Colasante A, Mascetra N, Piantelli M, Musiani P, Aiello FB (1998) IL-10 synergizes with dexamethasone in inhibiting human T cell proliferation. J Pharmacol Exp Ther 285:915–919
Buick RN, Pullano R, Trent JM (1985) Comparative properties of five human ovarian adenocarcinoma cell lines. Cancer Res 45:3668–3676
Curiel TJ, Coukos G, Zou L, Alvarez X, Cheng P, Mottram P, Evdemon-Hogan M, Conejo-Garcia JR, Zhang L, Burow M, Zhu Y, Wei S, Kryczek I, Daniel B, Gordon A, Myers L, Lackner A, Disis ML, Knutson KL, Chen L, Zou W (2004) Specific recruitment of regulatory T cells in ovarian carcinoma fosters immune privilege and predicts reduced survival. Nat Med 10:942–949
De Plaen E, Arden K, Traversari C, Gaforio JJ, Szikora JP, De Smet C, Brasseur F, van der Bruggen P, Lethe B, Lurquin C et al (1994) Structure, chromosomal localization, and expression of 12 genes of the MAGE family. Immunogenetics 40:360–369
De Smet C, Lurquin C, Van der Bruggen P, De Plaen E, Brasseur F, Boon T (1994) Sequence and expression pattern of the human MAGE2 gene. Immunogenetics 39:121–129
Disis ML, Smith JW, Murphy AE, Chen W, Cheever MA (1994) In vitro generation of human cytolytic T-cells specific for peptides derived from the HER-2/neu protooncogene protein. Cancer Res 54:1071–1076
Ellis SA, Taylor C, McMichael A (1982) Recognition of HLA-B27 and related antigen by a monoclonal antibody. Hum Immunol 5:49–59
Fisk B, Blevins TL, Wharton JT, Ioannides CG (1995) Identification of an immunodominant peptide of HER-2/neu protooncogene recognized by ovarian tumor-specific cytotoxic T lymphocyte lines. J Exp Med 181:2109–2117
Fogh J, Tremple G (1975) New human tumor cell lines. In: Fogh J (ed) Human tumor cell lines in vitro. Plenum Press, New York, pp 115–141
Fogh J, Wright WC, Loveless JD (1977) Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J Natl Cancer Inst 58:209–214
Furlong MT, Hough CD, Sherman-Baust CA, Pizer ES, Morin PJ (1999) Evidence for the colonic origin of ovarian cancer cell line SW626. J Natl Cancer Inst 91:1327–1328
Gillespie AM, Rodgers S, Wilson AP, Tidy J, Rees RC, Coleman RE, Murray AK (1998) MAGE, BAGE and GAGE: tumour antigen expression in benign and malignant ovarian tissue. Br J Cancer 78:816–821
Gordinier ME, Zhang HZ, Patenia R, Levy LB, Atkinson EN, Nash MA, Katz RL, Platsoucas CD, Freedman RS (1999) Quantitative analysis of transforming growth factor β1 and 2 in ovarian carcinoma. Clin Cancer Res 5:2498–2505
Gotlieb WH, Abrams JS, Watson JM, Velu TJ, Berek JS, Martinez-Maza O (1992) Presence of interleukin 10 (IL-10) in the ascites of patients with ovarian and other intra-abdominal cancers. Cytokine 4:385–390
Hamilton TC, Young RC, McKoy WM, Grotzinger KR, Green JA, Chu EW, Whang-Peng J, Rogan AM, Green WR, Ozols RF (1983) Characterization of a human ovarian carcinoma cell line (NIH:OVCAR-3) with androgen and estrogen receptors. Cancer Res 43:5379–5389
Haskill S, Becker S, Fowler W, Walton L (1982) Mononuclear-cell infiltration in ovarian cancer. I. Inflammatory-cell infiltrates from tumour and ascites material. Br J Cancer 45:728–736
Hogan KT, Coppola MA, Gatlin CL, Thompson LW, Shabanowitz J, Hunt DF, Engelhard VH, Ross MM, Slingluff CL (2004) Identification of novel and widely expressed cancer/testis gene isoforms that elicit spontaneous cytotoxic T lymphocyte reactivity to melanoma. Cancer Res 64:1157–1163
Jager D, Jager E, Knuth A (2001) Immune responses to tumour antigens: Implications for antigen specific immunotherapy of cancer. J Clin Pathol 54:669–674
Kabawat SE, Bast RC Jr, Welch WR, Knapp RC, Bhan AK (1983) Expression of major histocompatibility antigens and nature of inflammatory cellular infiltrate in ovarian neoplasms. Int J Cancer 32:547–554
Karlan BY, Amin W, Band V, Zurawski VR, Littlefield BA (1988) Plasminogen activator secretion by established lines of human ovarian carcinoma cells in vitro. Gynecol Oncol 31:103–112
Kim DK, Lee TV, Castilleja A, Anderson BW, Peoples GE, Kudelka AP, Murray JL, Sittisomwong T, Wharton JT, Kim JW, Ioannides CG (1999) Folate binding protein peptide 191–199 presented on dendritic cells can stimulate CTL from ovarian and breast cancer patients. Anticancer Res 19:2907–2916
Kuppen PJK, Schuitemaker H, van’t Veer LJ, de Bruijn EA, van Oosterom AT, Schrier PI (1988) Cis-diamminedichloroplatinum(II)-resistant sublines derived from two human ovarian tumor cell lines. Cancer Res 48:3355–3359
Lagendijk JH, Mullink H, Van Diest PJ, Meijer GA, Meijer CJLM (1998) Tracing the origin of adenocarcinomas with unknown primary using immunohistochemistry: differential diagnosis between colonic and ovarian carcinomas as primary sites. Hum Pathol 29:491–497
Lau DHM, Lewis AD, Ehsan MN, Sikic BI (1991) Multifactorial mechanisms associated with broad cross-resistance of ovarian carcinoma cells selected by cyanomorpholino doxorubicin. Cancer Res 51:5181–5187
Maier JA, Voulalas P, Roeder D, Maciag T (1990) Extension of the life-span of human endothelial cells by an interleukin-1 alpha antisense oligomer. Science 249:1570–1574
Makin CA, Bobrow LG, Bodmer WF (1984) Monoclonal antibody to cytokeratin for use in routine histopathology. J Clin Pathol 37:975–983
Moll R, Franke WW, Schiller DL (1982) The catalog of human cytokeratins: Patterns of expression in normal epithelia, tumors, and cultured cells. Cell 31:11–24
Mule JJ, Schwarz SL, Roberts AB, Sporn MB, Rosenberg SA (1988) Transforming growth factor-beta inhibits the in vitro generation of lymphokine-activated killer cells and cytotoxic T cells. Cancer Immunol Immunother 26:95–100
Nash MA, Lenzi R, Edwards CL, Kavanagh JJ, Kudelka AP, Verschraegen CF, Platsoucas CD, Freedman RS (1998) Differential expression of cytokine transcripts in human epithelial ovarian carcinoma by solid tumour specimens, peritoneal exudate cells containing tumour, tumour-infiltrating lymphocyte (TIL)-derived T cell lines and established tumour cell lines. Clin Exp Immunol 112:172–180
Negus RP, Stamp GW, Hadley J, Balkwill FR (1997) Quantitative assessment of the leukocyte infiltrate in ovarian cancer and its relationship to the expression of C-C chemokines. Am J Pathol 150:1723–1734
Novellino L, Castelli C, Parmiani G (2005) A listing of human tumor antigens recognized by T cells: March 2004 update. Cancer Immunol Immunother 54:187–207
Odunsi K, Jungbluth AA, Stockert E, Qian F, Gnjatic S, Tammela J, Intengan M, Beck A, Keitz B, Santiago D, Williamson B, Scanlan MJ, Ritter G, Chen Y-T, Driscoll D, Sood A, Lele S, Old LJ (2003) NY-ESO-1 and LAGE-1 cancer-testis antigens are potential targets for immunotherapy in epithelial ovarian cancer. Cancer Res 63:6076–6083
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
Parham P, Brodsky FM (1981) Partial purification and some properties of BB7.2. A cytotoxic monoclonal antibody with specificity for HLA-A2 and a variant of HLA-A28. Hum Immunol 3:277–299
Parmiani G, Castelli C, Dalerba P, Mortarini R, Rivoltini L, Marincola FM, Anichini A (2002) Cancer immunotherapy with peptide-based vaccines: What have we achieved? Where are we going? J Natl Cancer Inst 94:805–818
Peoples GE, Anderson BW, Fisk B, Kudelka AP, Wharton JT, Ioannides CG (1998) Ovarian cancer-associated lymphocyte recognition of folate binding protein peptides. Ann Surg Oncol 5:743–750
Peoples GE, Anderson BW, Lee TV, Murray JL, Kudelka AP, Wharton JT, Ioannides CG (1999) Vaccine implications of folate binding protein, a novel cytotoxic T lymphocyte-recognized antigen system in epithelial cancers. Clin Cancer Res 5:4214–4223
Peoples GE, Goedegebuure PS, Smith R, Linehan DC, Yoshino I, Eberlein TJ (1995) Breast and ovarian cancer-specific cytotoxic T lymphocytes recognize the same HER2/neu-derived peptide. Proc Natl Acad Sci U S A 92:432–436
Platsoucas CD, Fincke JE, Pappas J, Jung WJ, Heckel M, Schwarting R, Magira E, Monos D, Freedman RS (2003) Immune responses to human tumors: development of tumor vaccines. Anticancer Res 23:1969–1996
Provencher DM, Lounis H, Champoux L, Tetrault M, Manderson EN, Wang JC, Eydoux P, Savoie R, Tonin PN, Mes-Masson AM, Provencher DM, Lounis H, Champoux L, Tetrault M, Manderson EN, Wang JC, Eydoux P, Savoie R, Tonin PN, Mes-Masson AM (2000) Characterization of four novel epithelial ovarian cancer cell lines. In Vitro Cell Dev Biol Anim 36:357–361
Ranges GE, Figari IS, Espevik T, Palladino MA (1987) Inhibition of cytotoxic T cell development by transforming growth factor β and reversal by recombinant tumor necrosis factor α. J Exp Med 166:991–998
Rimoldi D, Salvi S, Schultz-Thater E, Spagnoli GC, Cerottini JC (2000) Anti-MAGE-3 antibody 57B and anti-MAGE-1 antibody 6C1 can be used to study different proteins in the MAGE-A family. Int J Cancer 86:749–751
Rosenberg SA (1999) A new era for cancer immunotherapy based on the genes that encode cancer antigens. Immunity 10:281–287
Russo C, Ng AK, Pellegrino MA, Ferrone S (1983) The monoclonal antibody CR11-351 discriminates HLA-A2 variants identified by T cells. Immunogenetics 18:23–35
Saalbach A, Anderegg U, Bruns M, Schnabel E, Herrmann K, Haustein UF (1996) Novel fibroblast-specific monoclonal antibodies: properties and specificities. J Investig Dermatol 106:1314–1319
Saalbach A, Aust G, Herrmann K, Anderegg U (1997) The fibroblast-specific mab AS02: a novel tool for detection and elimination of human fibroblasts. Cell Tissue Res 290:593–599
Santin AD, Bellone S, Ravaggi A, Roman J, Smith CV, Pecorelli S, Cannon MJ, Parham GP (2001) Increased levels of interleukin-10 and transforming growth factor-β in the plasma and ascitic fluid of patients with advanced ovarian cancer. Br J Obstet Gynaecol 108:804–808
Santin AD, Hermonat PL, Ravaggi A, Bellone S, Roman JJ, Smith CV, Pecorelli S, Radominska-Pandya A, Cannon MJ, Parham GP (2001) Phenotypic and functional analysis of tumor-infiltrating lymphocytes compared with tumor-associated lymphocytes from ascitic fluid and peripheral blood lymphocytes in patients with advanced ovarian cancer. Gynecol Obstet Invest 51:254–261
Seliger B, Cabrera T, Garrido F, Ferrone S (2002) HLA class I antigen abnormalities and immune escape by malignant cells. Semin Cancer Biol 12:3–13
Slingluff CL Jr., Hunt DF, Engelhard VH (1994) Direct analysis of tumor-associated peptide antigens. Curr Opin Immunol 6:733–740
Smedts F, Ramaekers F, Robben H, Pruszczynski Van Muijen G, Lane B, Leigh I, Vooijs P (1990) Changing patterns of keratin expression during progression of cervical intraepithelial neoplasia. Am J Pathol 136:657–668
Stimpfl M, Schmid BC, Schiebel I, Tong D, Leodolter S, Obermair A, Zeillinger R (1999) Expression of mucins and cytokeratins in ovarian cancer cell lines. Cancer Lett 145:133–141
Thomas DA, Massague J (2005) TGF-β directly targets cytotoxic T cell functions during tumor evasion of immune surveillance. Cancer Cell 8:369–380
Toutirais O, Chartier P, Dubois D, Bouet F, Leveque J, Catros-Quemener V, Genetet N (2003) Constitutive expression of TGF-beta1, interleukin-6 and interleukin-8 by tumor cells as a major component of immune escape in human ovarian carcinoma. Eur Cytokine Netw 14:246–255
Wallick SC, Figari IS, Morris RE, Levinson AD, Palladino MA (1990) Immunoregulatory role of transforming growth factor β (TGF-β) in development of killer cells: comparison of active and latent TGF-β1. J Exp Med 172:1777–1784
Wang RF, Johnston SL, Zeng G, Topalian SL, Schwartzentruber DJ, Rosenberg SA (1998) A breast and melanoma-shared tumor antigen: T cell responses to antigenic peptides translated from different open reading frames. J Immunol 161:3598–3606
Yamada A, Kataoka A, Shichijo S, Kamura T, Imai Y, Nishida T, Itoh K (1995) Expression of MAGE-1, MAGE-2, MAGE-3/-6 and MAGE-4a/-4b genes in ovarian tumors. Int J Cancer 64:388–393
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
Acknowledgments
This work was supported by grant W81XWH-05-1-0012 from the United States Department of Defense to K. T. Hogan.
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Carr, T.M., Adair, S.J., Fink, M.J. et al. Immunological profiling of a panel of human ovarian cancer cell lines. Cancer Immunol Immunother 57, 31–42 (2008). https://doi.org/10.1007/s00262-007-0347-7
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DOI: https://doi.org/10.1007/s00262-007-0347-7