Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Chronic lymphocytic leukemia

Antibodies conjugated with viral antigens elicit a cytotoxic T cell response against primary CLL ex vivo

Leukemia volume 33, pages 88–98 (2019)Cite this article

Subjects

Abstract

Chronic lymphocytic leukemia (CLL) is the most frequent B cell malignancy in Caucasian adults. The therapeutic armamentarium against this incurable disease has recently seen a tremendous expansion with the introduction of specific pathway inhibitors and innovative immunotherapy. However, none of these approaches is curative and devoid of side effects. We have used B-cell-specific antibodies conjugated with antigens (AgAbs) of the Epstein–Barr virus (EBV) to efficiently expand memory CD4+ cytotoxic T lymphocytes (CTLs) that recognized viral epitopes in 12 treatment-naive patients with CLL. The AgAbs carried fragments from the EBNA3C EBV protein that is recognized by the large majority of the population. All CLL cells pulsed with EBNA3C–AgAbs elicited EBV-specific T cell responses, although the intensity varied across the patient collective. Interestingly, a large proportion of the EBV-specific CD4+ T cells expressed granzyme B (GrB), perforin, and CD107a, and killed CLL cells loaded with EBV antigens with high efficiency in the large majority of patients. The encouraging results from this preclinical ex vivo study suggest that AgAbs have the potential to redirect immune responses toward CLL cells in a high percentage of patients in vivo and warrant the inception of clinical trials.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Dreger P, Schetelig J, Andersen N, Corradini P, van Gelder M, Gribben J, et al. Managing high-risk CLL during transition to a new treatment era: stem cell transplantation or novel agents? Blood. 2014;124:3841–9.

    Article  CAS  Google Scholar 

  2. Mato A, Porter DL. A drive through cellular therapy for CLL in 2015: allogeneic cell transplantation and CARs. Blood. 2015;126:478–85.

    Article  CAS  Google Scholar 

  3. Yu X, Ilecka M, Bartlett EJ, Schneidt V, Bhat R, Mautner J, et al. Antigen-armed antibodies targeting B lymphoma cells effectively activate antigen-specific CD4+T cells. Blood. 2015;125:1601–10.

    Article  CAS  Google Scholar 

  4. Lunde E, Bogen B, Sandlie I. Immunoglobulin as a vehicle for foreign antigenic peptides immunogenic to T cells. Mol Immunol. 1997;34:1167–76.

    Article  CAS  Google Scholar 

  5. Lunde E, Munthe LA, Vabo A, Sandlie I, Bogen B. Antibodies engineered with IgD specificity efficiently deliver integrated T cell epitopes for antigen presentation by B cells. Nat Biotechnol. 1999;17:670–5.

    Article  CAS  Google Scholar 

  6. Kang TH, Ma B, Wang CN, Wu TC, Hung CF. Targeted coating with antigenic peptide renders tumor cells susceptible to CD8(+) T cell-mediated killing. Mol Ther. 2013;21:542–53.

    Article  CAS  Google Scholar 

  7. Gurer C, Strowig T, Brilot F, Pack M, Trumpfheller C, Arrey F, et al. Targeting the nuclear antigen I of Epstein-Barr virus to the human endocytic receptor DEC-205 stimulates protective T-cell responses. Blood. 2008;112:1231–9.

    Article  CAS  Google Scholar 

  8. Rasmussen IB, Lunde E, Michaelsen TE, Bogen B, Sandlie I. The principle of delivery of T cell epitopes to antigen-presenting cells applied to peptides from influenza virus, ovalbumin, and hen egg lysozyme: Implications for peptide vaccination. Proc Natl Acad Sci USA. 2001;98:10296–301.

    Article  CAS  Google Scholar 

  9. Bozzacco L, Trumpfheller C, Siegal FP, Mehandru S, Markowitz M, Carrington M, et al. DEC-205 receptor on dendritic cells mediates presentation of HIV gag protein to CD8+T cells in a spectrum of human MHC I haplotypes. Proc Natl Acad Sci USA. 2007;104:1289–94.

    Article  CAS  Google Scholar 

  10. Do Y, Koh H, Park CG, Dudziak D, Seo P, Mehandru S, et al. Targeting of LcrV virulence protein from Yersinia pestis to dendritic cells protects mice against pneumonic plague. Eur J Immunol. 2010;40:2791–6.

    Article  CAS  Google Scholar 

  11. Cohen JI. Epstein-Barr virus infection. New Engl J Med. 2000;343:481–92.

    Article  CAS  Google Scholar 

  12. Nikiforow S, Bottomly K, Miller G. CD4+T-cell effectors inhibit Epstein-Barr virus-induced B-cell proliferation. J Virol. 2001;75:3740–52.

    Article  CAS  Google Scholar 

  13. Long HM, Haigh TA, Gudgeon NH, Leen AM, Tsang CW, Brooks J, et al. CD4+T-cell responses to Epstein-Barr virus (EBV) latent-cycle antigens and the recognition of EBV-transformed lymphoblastoid cell lines. J Virol. 2005;79:4896–907.

    Article  CAS  Google Scholar 

  14. Ramsay AG, Johnson AJ, Lee AM, Gorgun G, Le Dieu R, Blum W, et al. Chronic lymphocytic leukemia T cells show impaired immunological synapse formation that can be reversed with an immunomodulating drug. J Clin Investig. 2008;118:2427–37.

    CAS  PubMed  Google Scholar 

  15. Ramsay AG, Gribben JG. Immune dysfunction in chronic lymphocytic leukemia T cells and lenalidomide as an immunomodulatory drug. Haematol Hematol J. 2009;94:1198–202.

    Article  CAS  Google Scholar 

  16. Brusa D, Serra S, Coscia M, Rossi D, D’Arena G, Laurenti L, et al. The PD-1/PD-L1 axis contributes to T-cell dysfunction in chronic lymphocytic leukemia. Haematologica. 2013;98:953–63.

    Article  CAS  Google Scholar 

  17. McClanahan F, Riches JC, Miller S, Ghazaly E, Day W, Capasso M, et al. PD-1/PD-L1 mediated T-cell dysfunction in CLL is not absolute and can be at least partially reversed in vivo by the immune-modulatory drug lenalidomide. Oncol Res Treat. 2014;37:236–236.

    Google Scholar 

  18. Hofland H, de Weerdt W, Terpstra S, Remmerswaal EBM, ten Berge IJM, Kater AP, et al. EBV-specific CD8+T-cells are not functionally impaired in chronic lymphocytic leukemia. Blood. 2015;126:1723.

    Article  Google Scholar 

  19. Porakishvili N, Kardava L, Jewell AP, Yong K, Glennie MJ, Akbar A, et al. Cytotoxic CD4(+) T cells in patients with B cell chronic lymphocytic leukemia kill via a perforin-mediated pathway. Haematologica. 2004;89:435–43.

    CAS  PubMed  Google Scholar 

  20. Porakishvili N, Roschupkina T, Kalber T, Jewell AP, Patterson K, Yong K, et al. Expansion of CD4(+) T cells with a cytotoxic phenotype in patients with B-chronic lymphocytic leukaemia (B-CLL). Clin Exp Immunol. 2001;126:29–36.

    Article  CAS  Google Scholar 

  21. Simonetti G, Bertilaccio MTS, Ghia P, Klein U. Mouse models in the study of chronic lymphocytic leukemia pathogenesis and therapy. Blood. 2014;124:1010–9.

    Article  CAS  Google Scholar 

  22. World Medical Association. World medical association Declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA. 2013;310:2191–4.

    Article  Google Scholar 

  23. Hislop AD, Taylor GS, Sauce D, Rickinson AB. Cellular responses to viral infection in humans: lessons from Epstein-Barr virus. Annu Rev Immunol. 2007;25:587–617.

    Article  CAS  Google Scholar 

  24. Takeuchi A, Saito T. CD4 CTL, a cytotoxic subset of CD4(+) T cells, their differentiation and function. Front Immunol. 2017;8:194.

    Article  Google Scholar 

  25. Chen M, Shirai M, Liu ZJ, Arichi T, Takahashi H, Nishioka M. Efficient class II major histocompatibility complex presentation of endogenously synthesized hepatitis c virus core protein by Epstein-Barr virus-transformed B-lymphoblastoid cell lines to CD4(+) T cells. J Virol. 1998;72:8301–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Purner MB, Berens RL, Krug EC, Curiel TJ. Epstein-Barr virus-transformed B-cells, a potentially convenient source of autologous antigen-presenting cells for the propagation of certain human cytotoxic T-lymphocytes. Clin Diagn Lab Immunol. 1994;1:696–700.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Dazzi F, D’Andrea E, Biasi G, De Silvestro G, Gaidano G, Schena M, et al. Failure of B cells of chronic lymphocytic leukemia in presenting soluble and alloantigens. Clin Immunol Immunopathol. 1995;75:26–32.

    Article  CAS  Google Scholar 

  28. Bollard CM, Rooney CM, Heslop HE. T-cell therapy in the treatment of post-transplant lymphoproliferative disease. Nat Rev Clin Oncol. 2012;9:510–9.

    Article  CAS  Google Scholar 

  29. Gulley JL, Madan RA, Pachynski R, Mulders P, Sheikh NA, Trager J, et al. Role of antigen spread and distinctive characteristics of immunotherapy in cancer treatment. J Natl Cancer Inst. 2017;109.

  30. Hodgson K, Ferrer G, Montserrat E, Moreno C. Chronic lymphocytic leukemia and autoimmunity: a systematic review. Haematol Hematol J. 2011;96:752–61.

    Article  CAS  Google Scholar 

  31. Tandra P, Krishnamurthy J, Bhatt VR, Newman K, Armitage JO, Akhtari M. Autoimmune cytopenias in chronic lymphocytic leukemia, facts and myths. Mediterr J Hematol Infect Dis. 2013;5:e2013068.

    Article  Google Scholar 

  32. Merlo A, Turrini R, Bobisse S, Zamarchi R, Alaggio R, Dolcetti R, et al. Virus-specific cytotoxic CD4(+) T cells for the treatment of EBV-related tumors. J Immunol. 2010;184:5895–902.

    Article  CAS  Google Scholar 

  33. Mautner J, Bornkamm GW. The role of virus-specific CD4+T cells in the control of Epstein-Barr virus infection. Eur J Cell Biol. 2012;91:31–35.

    Article  CAS  Google Scholar 

  34. Derby EG, Reddy V, Nelson EL, Kopp WC, Baseler MW, Dawson JR, et al. Correlation of human CD56+cell cytotoxicity and IFN-gamma production. Cytokine. 2001;13:85–90.

    Article  CAS  Google Scholar 

  35. Horton H, Russell N, Moore E, Frank I, Baydo R, Havenar-Daughton C, et al. Correlation between interferon-gamma secretion and cytotoxicity, in virus-specific memory T cells. J Infect Dis. 2004;190:1692–6.

    Article  CAS  Google Scholar 

  36. Spender LC, Cannell EJ, Hollyoake M, Wensing B, Gawn JM, Brimmell M, et al. Control of cell cycle entry and apoptosis in B lymphocytes infected by Epstein-Barr virus. J Virol. 1999;73:4678–88.

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Godal R, Keilholz U, Uharek L, Letsch A, Asemissen AM, Busse A, et al. Lymphomas are sensitive to perforin-dependent cytotoxic pathways despite expression of PI-9 and overexpression of bcl-2. Blood. 2006;107:3205–11.

    Article  CAS  Google Scholar 

  38. Vanura K, Rieder F, Kastner MT, Biebl J, Sandhofer M, Le T, et al. Chronic lymphocytic leukemia patients have a preserved cytomegalovirus-specific antibody response despite progressive hypogammaglobulinemia. PLoS ONE. 2013;8:e78925.

    Article  CAS  Google Scholar 

  39. Saxena A, Wu DH. Advances in therapeutic Fc engineering - modulation of igG-associated effector functions and serum half-life. Front Immunol. 2016;7:580.

    Article  Google Scholar 

  40. Martelius T, Lappalainen M, Aalto SM, Nihtinen A, Hedman K, Anttila VJ. Clinical characteristics, outcome and the role of viral load in nontransplant patients with Epstein-Barr viraemia. Clin Microbiol Infect. 2010;16:657–62.

    Article  CAS  Google Scholar 

  41. Dolcetti R, Carbone A. Epstein-Barr virus infection and chronic lymphocytic leukemia: a possible progression factor? Infect Agent Cancer. 2010;5:22.

    Article  Google Scholar 

  42. Lazzarino M, Orlandi E, Baldanti F, Furione M, Pagnucco G, Astori C, et al. The immunosuppression and potential for EBV reactivation of fludarabine combined with cyclophosphamide and dexamethasone in patients with lymphoproliferative disorders. Br J Haematol. 1999;107:877–82.

    Article  CAS  Google Scholar 

  43. te Raa GD, Pascutti MF, García-Vallejo JJ, Reinen E, Remmerswaal EB, ten Berge IJ, et al. CMV-specific CD8+T-cell function is not impaired in chronic lymphocytic leukemia. Blood. 2014;123:717–24.

    Article  Google Scholar 

  44. Rezvani AR, Maloney DG. Rituximab resistance. Best Pract Res Clin Haematol. 2011;24:203–16.

    Article  CAS  Google Scholar 

  45. Taylor RP, Lindorfer MA. Antigenic modulation and rituximab resistance. Semin Hematol. 2010;47:124–32.

    Article  CAS  Google Scholar 

  46. Lim SH, Vaughan AT, Ashton-Key M, Williams EL, Dixon SV, Chan HTC, et al. Fc gamma receptor IIb on target B cells promotes rituximab internalization and reduces clinical efficacy. Blood. 2011;118:2530–40.

    Article  Google Scholar 

  47. Beers SA, French RR, Chan HT, Lim SH, Jarrett TC, Vidal RM, et al. Antigenic modulation limits the efficacy of anti-CD20 antibodies: implications for antibody selection. Blood. 2010;115:5191–201.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the German Cancer Aid (Deutsche Krebshilfe) (VS, MI, and DGvZ). The authors thank all participants, patients, and healthy volunteers, as well as physicians, nurses, and clinical investigators. We thank Martin Müller, PhD (DKFZ, F035), Remy Poirey, PhD, and Zhe Li (DKFZ, F100) for helpful discussions.

Author contributions

VS performed the experiments, analyzed the data, and wrote the manuscript. MI and DGvZ undertook research and provided technical support. PD designed the clinical protocol and provided blood samples. SF designed the clinical investigation protocol and undertook research. JM provided T cell clones and oversaw research. HJD planned the study, oversaw the research, and wrote the manuscript. All authors approved the article.

Funding

This work was supported by a grant from the German Cancer Aid (Deutsche Krebshilfe).

Author information

Authors and Affiliations

  1. German Cancer Research Center (DKFZ), Unit F100, 69120, Heidelberg, Germany

    Viktor Schneidt, Marta Ilecka, Dwain G. van Zyl, Susanne Fink & Henri-Jacques Delecluse

  2. Faculty of Biosciences, Heidelberg University, 69120, Heidelberg, Germany

    Viktor Schneidt, Dwain G. van Zyl & Henri-Jacques Delecluse

  3. Institut National de la Santé et de la Recherche Médicale (INSERM) Unit U1074, DKFZ, 69120, Heidelberg, Germany

    Viktor Schneidt, Marta Ilecka, Dwain G. van Zyl, Susanne Fink & Henri-Jacques Delecluse

  4. German Center for Infection Research (DZIF), 69120, Heidelberg, Germany

    Viktor Schneidt, Marta Ilecka, Dwain G. van Zyl, Susanne Fink & Henri-Jacques Delecluse

  5. Department of Internal Medicine V, Heidelberg University Hospital, 69120, Heidelberg, Germany

    Peter Dreger

  6. Nierenzentrum Heidelberg, 69120, Heidelberg, Germany

    Susanne Fink

  7. Research Unit Gene Vectors, Helmholtz Zentrum München, 81377, Munich, Germany

    Josef Mautner

  8. Children’s Hospital Technische Universität München, 80804, Munich, Germany

    Josef Mautner

  9. DZIF, 81377, Munich, Germany

    Josef Mautner

Authors
  1. Viktor Schneidt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marta Ilecka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Peter Dreger
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dwain G. van Zyl
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Susanne Fink
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Josef Mautner
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Henri-Jacques Delecluse
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Henri-Jacques Delecluse.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Schneidt, V., Ilecka, M., Dreger, P. et al. Antibodies conjugated with viral antigens elicit a cytotoxic T cell response against primary CLL ex vivo. Leukemia 33, 88–98 (2019). https://doi.org/10.1038/s41375-018-0160-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41375-018-0160-7

Search

Advanced search

Quick links