Skip to main content

Advertisement

SpringerLink
Log in

Dendritic cell-based tumor vaccine for cervical cancer I: in vitro stimulation with recombinant protein-pulsed dendritic cells induces specific T cells to HPV16 E7 or HPV18 E7

  • Original Paper
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Purpose

Human papillomavirus (HPV) type 16 and 18 are the most prevalent genotypes in cervical cancers. The viral oncoproteins E6 and E7 are considered to be tumor-specific targets for immunotherapy. HPV E7 antigen-loaded dendritic cells (DC) were evaluated as cellular tumor vaccine.

Methods

Autologous monocyte-derived DCs loaded with recombinant HPV16 or HPV18 E7 oncoprotein were used to induce in vitro a specific T cell response. Specificities of activated T cells were determined.

Results

E7-specific T cells could be identified in 18/20 T cell lines from healthy blood donors. CD4+ T cell responses (13/16) were found by proliferation assay. CD8+ CTLs (12/18) were detectable by interferon-gamma (IFN-γ) ELISpot analysis. Seven donors reacted in both assays and only 2/20 T cell lines did not react in any assay. Thus, specific T cells could be activated in >80% of healthy individuals. T cell lines from suitable donors were specific for HLA-A*0201-restricted epitopes. Furthermore, HPV E7 antigen-loaded DC stimulated specific responses in freshly isolated tumor infiltrating lymphocyte (TIL) populations of cervical cancer patients.

Conclusion

Autologous dendritic cells loaded with HPV E7 protein can induce T cell responses in healthy individuals by in vitro stimulation and evoke responses in TIL from cervical cancer biopsies. Since there are no limitations with respect to specific HLA-haplotypes, these findings may be a basis for the development of a therapeutic protein-based DC tumor vaccine against cervical cancer for HPV16- and HPV18-positive patients.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig.1.
Fig. 2a,b.
Fig. 3a,b.
Fig. 4.
Fig. 5a,b.

Similar content being viewed by others

Abbreviations

B-LCL :

B lymphoblastoid cell line

CTL :

Cytotoxic T lymphocyte

DC :

Dendritic cell

IFN-γ :

Interferon-gamma

IL :

Interleukin

GM-CSF :

Granulocyte/macrophage-colony stimulating factor

HPV :

Human Papillomavirus

HSA :

human serum albumin

PBL :

Peripheral blood lymphocytes

PBMC :

Peripheral blood mononuclear cells

SD :

standart deviation

TIL :

Tumor infiltrating lymphocytes

TNF-α :

Tumor necrosis factor-alpha

References

  • Bosch FX, Manos MM, Munoz N, Sherman M, Jansen AM, Peto J, Schiffman MH, Moreno V, Kurman R, Shah KV (1995) Prevalence of human papillomavirus in cervical cancer: a worldwide perspective. International biological study on cervical cancer (IBSCC) Study Group. J Natl Cancer Inst 87:796–802

    CAS  PubMed  Google Scholar 

  • Braspenning J, Manetti R, Zumbach K, Meschede W, Gissmann L, Tommasino M (1997) A general purification protocol for E7 proteins from "high- and low-risk" human papillomavirus types expressed in the yeast Schizosaccharomyces pombe. Protein Expr Purif 10:192–201

    Article  CAS  PubMed  Google Scholar 

  • Brinton LA (1992) Epidemiology of cervical cancer–overview. IARC Sci Publ 119:3–23

    PubMed  Google Scholar 

  • Castellanos MR, Hayes RL, Maiman MA (2001) Synthetic peptides induce a cytotoxic response against human papillomavirus type-18. Gynecol Oncol 82:77–83

    Article  CAS  PubMed  Google Scholar 

  • Chen LP, Thomas EK, Hu SL, Hellstrom I, Hellstrom KE. (1991) Human papillomavirus 16 nucleoprotein E7 is a tumor rejection antigen. Proc Natl Acad Sci USA 88:110–114

    CAS  PubMed  Google Scholar 

  • Coleman N, Birley HD, Renton AM, Hanna NF, Ryait BK, Byrne M, Taylor-Robinson D, Stanley MA (1994) Immunological events in regressing genital warts. Am J Clin Pathol 102:768–774

    CAS  PubMed  Google Scholar 

  • De Bruijn ML, Schuurhuis DH, Vierboom MP, Vermeulen H, de Cock KA, Ooms ME, Ressing ME, Toebes M, Franken KL, Drijfhout JW, Ottenhoff TH, Offringa R, Melief CJ (1998) Immunization with human papillomavirus type 16 (HPV16) oncoprotein-loaded dendritic cells as well as protein in adjuvant induces MHC class I-restricted protection to HPV16- induced tumor cells. Cancer Res 58:724–731

    PubMed  Google Scholar 

  • de Jong A, van der Burg SH, Kwappenberg KM, van der Hulst JM, Franken KL, Geluk A, van Meijgaarden KE, Drijfhout JW, Kenter G, Vermeij P, Melief CJ, Offringa R (2002) Frequent detection of human papillomavirus 16 E2-specific T-helper immunity in healthy subjects. Cancer Res 62:472–479

    PubMed  Google Scholar 

  • Evans EM, Man S, Evans AS, Borysiewicz LK (1997) Infiltration of cervical cancer tissue with human papillomavirus-specific cytotoxic T-lymphocytes. Cancer Res 57:2943–2950

    CAS  PubMed  Google Scholar 

  • Feltkamp MCW, Smits HL, Vierboom MPM, Minnaar RP, de Jongh BM, Drijfhout JW, ter Schegget J, Melief CJM, Kast WM (1993) Vaccination with cytotoxic T lymphocyte epitope-containing peptide protects against a tumor induced by human papillomavirus type 16-transformed cells. Eur J Immunol 23:2242–2249

    CAS  PubMed  Google Scholar 

  • Friedl J, Stift A, Paolini P, Roth E, Steger GG, Mader R, Jakesz R, Gnant MF (2000) Tumor antigen pulsed dendritic cells enhance the cytolytic activity of tumor infiltrating lymphocytes in human hepatocellular cancer. Cancer Biother Radiopharm 15:477–486

    CAS  PubMed  Google Scholar 

  • Ho GY, Burk RD, Fleming I, Klein RS (1994) Risk of genital human papillomavirus infection in women with human immunodeficiency virus-induced immunosuppression. Int J Cancer 56:788–792

    CAS  PubMed  Google Scholar 

  • Honma S, Tsukada S, Honda S, Nakamura M, Takakuwa K, Maruhashi S, Kodama S, Konazawa K, Takahashi T, Tanaka K (1994) Biological-clinical significance of selective loss of HLA-class I allelic product expression in squamous-cell carcinoma of the uterine cervix. Int J Cancer 57:650–655

    CAS  PubMed  Google Scholar 

  • Jacobs MV, Snijders PJ, van den Brule AJ, Helmerhorst TJ, Meijer CJ, Walboomers JM (1997) A general primer GP5+/GP6(+)-mediated PCR-enzyme immunoassay method for rapid detection of 14 high-risk and 6 low-risk human papillomavirus genotypes in cervical scrapings. J Clin Microbiol 35:791–795

    CAS  PubMed  Google Scholar 

  • Jonuleit H, Kuhn U, Muller G, Steinbrink K, Paragnik L, Schmitt E, Knop J, Enk AH (1997) Pro-inflammatory cytokines and prostaglandins induce maturation of potent immunostimulatory dendritic cells under fetal calf serum-free conditions. Eur J Immunol 27:3135–3142

    CAS  PubMed  Google Scholar 

  • Kather A, Ferrara A, Nonn M, Schinz M, Nieland J, Schneider A, Dürst M, Kaufmann AM (2003) Identification of a naturally processed HLA-A*0201 HPV18 E7 T cell epitope by tumor cell mediated in vitro vaccination. Int J Cancer 104:345–353

    Article  CAS  PubMed  Google Scholar 

  • Kaufmann AM, Gissmann L, Schreckenberger C, Qiao L (1997) Cervical carcinoma cells transfected with the CD80 gene elicit a primary cytotoxic T lymphocyte response specific for HPV 16 E7 antigens. Cancer Gene Ther4:377–382

    Google Scholar 

  • Kaufmann AM, Gissmann l, Simms P, Schreckenberger C, Qiao L (2000) Comparison of cytokines and CD80 for enhancement of immunogenicity of cervical cancer cells. Immunobiol 202:339–352

    CAS  Google Scholar 

  • Kaufmann AM, Nieland J, Schinz M, Nonn M, Gabelsberger J, Meissner H, Muller RT, Jochmus I, Gissmann L, Schneider A, Durst M (2001) HPV16 L1E7 chimeric virus-like particles induce specific HLA-restricted T cells in humans after in vitro vaccination. Int J Cancer 92:285–293

    Article  CAS  PubMed  Google Scholar 

  • Liu Y, Chiriva-Internati M, Grizzi F, Salati E, Roman JJ, Lim S, Hermonat PL (2001) Rapid induction of cytotoxic T cell response against cervical cancer cells by human papillomavirus type 16 E6 antigen gene delivery into human dendritic cells by an adeno-associated virus vector. Cancer Gene Ther 8:948–957

    Article  CAS  PubMed  Google Scholar 

  • Meschede W, Zumbach K, Braspenning J, Scheffner M, Benitez-Bribiesca L, Luande J, Gissmann L, Pawlita M (1998) Antibodies against early proteins of human papillomaviruses as diagnostic markers for invasive cervical cancer. J Clin Microbiol 36:475–480

    CAS  PubMed  Google Scholar 

  • Michel N, Öhlschläger P, Osen W, Freyschmidt EJ, Guthörlein H, Kaufmann A, Müller M, Gissmann L (2002) T cell response against human papillomavirus (HPV) 16 E7 in mice: comparison of Cr-release assay, intracellular IFN-γ production, ELISPOT and tetramer staining. Intervirology 45:290–299

    Google Scholar 

  • Mulders P, Tso CL, Gitlitz B, Kaboo R, Hinkel A, Frand S, Kiertscher S, Roth MD, deKernion J, Figlin R, Belldegrun A (1999) Presentation of renal tumor antigens by human dendritic cells activates tumor-infiltrating lymphocytes against autologous tumor: implications for live kidney cancer vaccines. Clin Cancer Res 5:445–454

    CAS  PubMed  Google Scholar 

  • Munderspach L, Wilczynski S, Roman L, Bade L, Felix J, Small LA, Kast MW, Fascio G, Marty V, Weber J (2000) A phase I trial of a human papillomavirus (HPV) peptide vaccine for women with high-grade cervical and vulvar intraepithelial neoplasia who are HPV 16 positive. Clin Canc Res 6:3406–3416

    Google Scholar 

  • Murakami M, Gurski KJ, Marincola FM, Ackland J, Steller MA (1999) Induction of specific CD8+ T-lymphocyte responses using a human papillomavirus-16 E6/E7 fusion protein and autologous dendritic cells. Cancer Res 59:1184–1187

    CAS  PubMed  Google Scholar 

  • Nestle FO, Alijagic S, Gilliet M, Sun Y, Grabbe S, Dummer R, Burg G, Schadendorf D (1998) Vaccination of melanoma patients with peptide-or tumor lysate-pulsed dendritic cells. Nat Med 4:328–332

    CAS  PubMed  Google Scholar 

  • Nikoloff B, Turka L (1994) Immunological functions of nonprofessional antigen-presenting cells: new insights from studies of T-cell interaction with keratinocytes. Immunol Today 15:464–469

    Article  PubMed  Google Scholar 

  • Nindl I, Lotz B, Kuhne-Heid R, Endisch U, Schneider A (1999) Distribution of 14 high risk HPV types in cervical intraepithelial neoplasia detected by a non-radioactive general primer PCR mediated enzyme immunoassay. J Clin Pathol 52:17–22

    CAS  PubMed  Google Scholar 

  • Ossevoort MA, Feltkamp MC, van Veen KJ, Melief CJ, Kast WM (1995) Dendritic cells as carriers for a cytotoxic T-lymphocyte epitope-based peptide vaccine in protection against a human papillomavirus type 16-induced tumor. J Immunother Emphasis Tumor Immunol 18:86–94

    CAS  PubMed  Google Scholar 

  • Petry KU, Scheffel D, Bode U, Gabrysiak T, Kochel H, Kupsch E, Glaubitz M, Niesert S, Kuhnle H, Schedel I (1994) Cellular immunodeficiency enhances the progression of human papillomavirus-associated cervical lesions. Int J Cancer 57:836–840

    CAS  PubMed  Google Scholar 

  • Ressing ME, Sette A, Brandt RMP, Ruppert J, Wentworth PA, Hartman M, Oseroff C, Grey HM, Melief CJM, Kast WM (1995) Human CTL epitopes encoded by human papillomavirus type 16 E6 and E7 identified through in vivo and in vitro immunogenicity studies of HLA-A*0201-binding peptides. J Immunol 154:5934–5943

    CAS  PubMed  Google Scholar 

  • Ressing ME, van Driel WJ, Brandt RM, Kenter GG, de Jong JH, Bauknecht T, Fleuren GJ, Hoogerhout P, Offringa R, Sette A, Celis E, Grey H, Trimbos BJ, Kast WM, Melief CJ (2000) Detection of T helper responses, but not of human papillomavirus-specific cytotoxic T lymphocyte responses, after peptide vaccination of patients with cervical carcinoma. J Immunother 23:255–266

    Article  CAS  PubMed  Google Scholar 

  • Rodriguez A, Regnault A, Kleijmeer M, Ricciardi-Castagnoli P, Amigorena S (1999) Selective transport of internalized antigens to the cytosol for MHC class I presentation in dendritic cells. Nat Cell Biol 1:362–368

    Article  CAS  PubMed  Google Scholar 

  • Romani N, Gruner S, Brang D, Kampgen E, Lenz A, Trockenbacher B, Konwalinka G, Fritsch PO, Steinman RM, Schuler G (1994) Proliferating dendritic cell progenitors in human blood. J Exp Med 180:83–93

    CAS  PubMed  Google Scholar 

  • Romani N, Reider D, Heuer M, Ebner S, Kampgen E, Eibl B, Niederwieser D, Schuler G (1996) Generation of mature dendritic cells from human blood. An improved method with special regard to clinical applicability. J Immunol Methods 196:137–151

    CAS  PubMed  Google Scholar 

  • Rudolf MP, Man S, Melief CJ, Sette A, Kast WM (2001) Human T cell responses to HLA-A-restricted high binding affinity peptides of human papillomavirus type 18 proteins E6 and E7. Clin Cancer Res 7:788s–795s

    CAS  PubMed  Google Scholar 

  • Sallusto F, Cella M, Danieli C, Lanzavecchia A (1995) Dendritic cells use macropinocytosis and the mannose receptor to concentrate macromolecules in the major histocompatibility complex class II compartment: downregulation by cytokines and bacterial products. J Exp Med 182:389–400

    CAS  PubMed  Google Scholar 

  • Santin AD, Hermonat PL, Ravaggi A, Chiriva-Internati M, Zhan D, Pecorelli S, Parham GP, Cannon MJ (1999) Induction of human papillomavirus-specific CD4(+) and CD8(+) lymphocytes by E7-pulsed autologous dendritic cells in patients with human papillomavirus type 16- and 18-positive cervical cancer. J Virol 73:5402–5410

    CAS  PubMed  Google Scholar 

  • Santin AD, Hermonat PL, Ravaggi A, Bellone S, Roman JJ, Jayaprabhu S, Pecorelli S, Parham GP, Cannon MJ (2001) Expression of CD56 by human papillomavirus E7-specific CD8+ cytotoxic T lymphocytes correlates with increased intracellular perforin expression and enhanced cytotoxicity against HLA-A2-matched cervical tumor cells. Clin Cancer Res 7:804s–810s

    CAS  PubMed  Google Scholar 

  • Schuler G, Steinman RM (1997) Dendritic cells as adjuvants for immune-mediated resistance to tumors. J Exp Med 186:1183–1187

    CAS  PubMed  Google Scholar 

  • Steinman RM (1991) The dendritic cell system and its role in immunogenicity. Annu Rev Immunol 9:271–296

    PubMed  Google Scholar 

  • Stoler M, Rhodes C, Whitbeck A, Wolinsky S, Chow L, Brocker T (1992) Human papillomavirus type 16 and 18 gene expression in cervical neoplasias. Hum Pathol 23:117–128

    CAS  PubMed  Google Scholar 

  • Sulitzeanu D (1993) Immunosuppressive factors in human cancer. Adv Cancer Res 60:247–267

    CAS  PubMed  Google Scholar 

  • Thurner B, Roder C, Dieckmann D, Heuer M, Kruse M, Glaser A, Keikavoussi P, Kampgen E, Bender A, Schuler G (1999) Generation of large numbers of fully mature and stable dendritic cells from leukapheresis products for clinical application. J Immunol Methods 223:1–15

    CAS  PubMed  Google Scholar 

  • Tuting T, DeLeo AB, Lotze MT, Storkus WJ (1997) Genetically modified bone marrow-derived dendritic cells expressing tumor-associated viral or "self" antigens induce antitumor immunity in vivo. Eur J Immunol 27:2702–2707

    CAS  PubMed  Google Scholar 

  • von Knebel-Doeberitz M, Oltersdorf T, Schwarz E, Gissmann L (1988) Correlation of modified human papilloma virus early gene expression with altered growth properties in C4–1 cervical carcinoma cells. Cancer Res 48:3780–3786

    PubMed  Google Scholar 

  • von Knebel-Doeberitz M, Rittmuller C, zur Hausen H, Durst M (1992) Inhibition of tumorigenicity of cervical cancer cells in nude mice by HPV E6-E7 anti-sense RNA. Int J Cancer 51:831–834

    PubMed  Google Scholar 

  • Walboomers JM, Jacobs MV, Manos MM, Bosch FX, Kummer JA, Shah KV, Snijders PJ, Peto J, Meijer CJ, Munoz N (1999) Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol 89:12–19

    Article  Google Scholar 

  • Welters MJP, de Jong A, van den Enden SJF, van der Hulst JP, Kwappenberg KMC, Hassane S, Franken KLMC, Drijfhout JW, Fleuren GJ, Kenter G, Melief CJM, Offringa R, van der Burg SH (2003) Frequent display of human papillomavirus type 16 E6-specific memory T-helper cells in the healthy population as witness of previous viral encounter. Cancer Res 63:636–641

    CAS  PubMed  Google Scholar 

  • Williams OM, Hart KW, Wang EC, Gelder CM (2002) Analysis of CD4(+) T-cell responses to human papillomavirus (HPV) type 11 L1 in healthy adults reveals a high degree of responsiveness and cross-reactivity with other HPV types. J Virol 76:7418–7429

    Article  CAS  PubMed  Google Scholar 

  • Wollscheid V, Kuhne-Heid R, Stein I, Jansen L, Kollner S, Schneider A, Durst M (2002) Identification of a new proliferation-associated protein NET-1/C4.8 characteristic for a subset of high-grade cervical intraepithelial neoplasia and cervical carcinomas. Int J Cancer 99:771–775

    Article  CAS  PubMed  Google Scholar 

  • Youde S, McCarthy C, Dunbar R, Cerundolo V, Borysiewicz LK, Man S (2000) Cross-typic specificity of human HPV16 E7 specific CTL lines and the relationship to TCR affinity. 18th International Papillomavirus Conference, 2000, Barcelona, Spain

  • zur Hausen H (1991) Human papillomaviruses in the pathogenesis of anogenital cancer. Virology 184:9–13

    PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported in part by grants SCHN 294/6–1 and SCHN 294/6–2 of the Deutsche Forschungsgemeinschaft and 01-ZZ-0602 by BMBF. The authors are grateful to Norman Häfner for sequence comparison of HPV16 and HPV18 E7 and Dr. C. Schreckenberger for statistical analysis and critical reading of the manuscript.

Author information

Authors and Affiliations

  1. Gynäkologische Molekularbiologie, Universitätsfrauenklinik, Friedrich-Schiller-Universität Jena, Bachstrasse 18, 07743, Jena, Germany

    Marion Nonn, Manuela Schinz, Achim Schneider, Matthias Dürst & Andreas M. Kaufmann

  2. Angewandte Tumorvirologie, Deutsches Krebsforschungszentrum, INF 242, 69120, Heidelberg, Germany

    Klaus Zumbach & Michael Pawlita

Authors
  1. Marion Nonn
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Manuela Schinz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Klaus Zumbach
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Michael Pawlita
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Achim Schneider
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Matthias Dürst
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Andreas M. Kaufmann
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Andreas M. Kaufmann.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nonn, M., Schinz, M., Zumbach, K. et al. Dendritic cell-based tumor vaccine for cervical cancer I: in vitro stimulation with recombinant protein-pulsed dendritic cells induces specific T cells to HPV16 E7 or HPV18 E7. J Cancer Res Clin Oncol 129, 511–520 (2003). https://doi.org/10.1007/s00432-003-0462-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00432-003-0462-6

Keywords

Search

Navigation