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.

  • Original Article
  • Published:

Therapy

A bispecific single-chain antibody that mediates target cell-restricted, supra-agonistic CD28 stimulation and killing of lymphoma cells

Leukemia volume 23, pages 71–77 (2009)Cite this article

Abstract

We have previously reported that r28M, a recombinant bispecific single-chain antibody directed to a melanoma-associated proteoglycan (NG2) and the costimulatory CD28 molecule on T cells, induced T-cell activation, which resulted in tumor-cell killing. T-cell activation did not require a primary signal through the T-cell antigen receptor (TCR)/CD3 complex and depended on the presence of NG2-positive tumor cells. Here, we further investigate this phenomenon of a target cell-restricted, supra-agonistic CD28 stimulation with bispecific antibodies. To this end, we exchanged the NG2 targeting part of r28M with a single-chain antibody directed to the B-cell associated antigen CD20. The resulting bispecific single-chain antibody, termed r2820, induced supra-agonistic T-cell activation, which required the presence of autologous normal or malignant B cells, respectively. Once activated, T cells were capable of destroying lymphoma target cells.

These findings demonstrate that supra-agonistic CD28 stimulation with bispecific single-chain antibodies is a robust and readily reproducible phenomenon. In the context of experimental tumor therapy, it may provide a valuable alternative to the unrestricted T-cell activation induced by ‘super-agonistic’, monospecific CD28 antibodies.

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Zang X, Allison JP . The B7 family and cancer therapy: costimulation and coinhibition. Clin Cancer Res 2007; 13 (18 Part 1): 5271–5279.

    Article  CAS  Google Scholar 

  2. Tacke M, Hanke G, Hanke T, Hunig T . CD28-mediated induction of proliferation in resting T cells in vitro and in vivo without engagement of the T cell receptor: evidence for functionally distinct forms of CD28. Eur J Immunol 1997; 27: 239–247.

    Article  CAS  Google Scholar 

  3. Elflein K, Rodriguez-Palmero M, Kerkau T, Hunig T . Rapid recovery from T lymphopenia by CD28 superagonist therapy. Blood 2003; 102: 1764–1770.

    Article  CAS  Google Scholar 

  4. Beyersdorf N, Hanke T, Kerkau T, Hunig T . Superagonistic anti-CD28 antibodies: potent activators of regulatory T cells for the therapy of autoimmune diseases. Ann Rheum Dis 2005; 64 (Suppl 4): iv91–iv95.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Suntharalingam G, Perry MR, Ward S, Brett SJ, Castello-Cortes A, Brunner MD et al. Cytokine storm in a phase 1 trial of the anti-CD28 monoclonal antibody TGN1412. N Engl J Med 2006; 355: 1018–1028.

    Article  CAS  Google Scholar 

  6. Grosse-Hovest L, Hartlapp I, Marwan W, Brem G, Rammensee HG, Jung G . A recombinant bispecific single-chain antibody induces targeted, supra-agonistic CD28-stimulation and tumor cell killing. Eur J Immunol 2003; 33: 1334–1340.

    Article  CAS  Google Scholar 

  7. Grosse-Hovest L, Muller S, Minoia R, Wolf E, Zakhartchenko V, Wenigerkind H et al. Cloned transgenic farm animals produce a bispecific antibody for T cell-mediated tumor cell killing. Proc Natl Acad Sci USA 2004; 101: 6858–6863.

    Article  CAS  Google Scholar 

  8. Grosse-Hovest L, Wick W, Minoia R, Weller M, Rammensee HG, Brem G et al. Supraagonistic, bispecific single-chain antibody purified from the serum of cloned, transgenic cows induces T-cell-mediated killing of glioblastoma cells in vitro and in vivo. Int J Cancer 2005; 117: 1060–1064.

    Article  CAS  Google Scholar 

  9. Liu AY, Robinson RR, Murray Jr ED, Ledbetter JA, Hellstrom I, Hellstrom KE . Production of a mouse-human chimeric monoclonal antibody to CD20 with potent Fc-dependent biologic activity. J Immunol 1987; 139: 3521–3526.

    CAS  PubMed  Google Scholar 

  10. Jung G, Freimann U, Von Marschall Z, Reisfeld RA, Wilmanns W . Target cell-induced T cell activation with bi- and trispecific antibody fragments. Eur J Immunol 1991; 21: 2431–2435.

    Article  CAS  Google Scholar 

  11. Cemerlic D, Dadey B, Han T, Vaickus L . Cytokine influence on killing of fresh chronic lymphocytic leukemia cells by human leukocytes. Blood 1991; 77: 2707–2715.

    CAS  PubMed  Google Scholar 

  12. Rossi E, Matutes E, Morilla R, Owusu-Ankomah K, Heffernan AM, Catovsky D . Zeta chain and CD28 are poorly expressed on T lymphocytes from chronic lymphocytic leukemia. Leukemia 1996; 10: 494–497.

    CAS  PubMed  Google Scholar 

  13. Frydecka I, Kosmaczewska A, Bocko D, Ciszak L, Wolowiec D, Kuliczkowski K et al. Alterations of the expression of T-cell-related costimulatory CD28 and downregulatory CD152 (CTLA-4) molecules in patients with B-cell chronic lymphocytic leukaemia. Br J Cancer 2004; 90: 2042–2048.

    Article  CAS  Google Scholar 

  14. Tacke M, Hanke G, Hanke T, Hunig T . CD28-mediated induction of proliferation in resting T cells in vitro and in vivo without engagement of the T cell receptor: evidence for functionally distinct forms of CD28. Eur J Immunol 1997; 27: 239–247.

    Article  CAS  Google Scholar 

  15. Kunicka JE, Platsoucas CD . Defective helper function of purified T4 cells and excessive suppressor activity of purified T8 cells in patients with B-cell chronic lymphocytic leukemia. T4 suppressor effector cells are present in certain patients. Blood 1988; 71: 1551–1560.

    CAS  PubMed  Google Scholar 

  16. Foon KA, Rai KR, Gale RP . Chronic lymphocytic leukemia: new insights into biology and therapy. Ann Intern Med 1990; 113: 525–539.

    Article  CAS  Google Scholar 

  17. Platsoucas CD, Galinski M, Kempin S, Reich L, Clarkson B, Good RA . Abnormal T lymphocyte subpopulations in patients with B cell chronic lymphocytic leukemia: an analysis by monoclonal antibodies. J Immunol 1982; 129: 2305–2312.

    CAS  PubMed  Google Scholar 

  18. Totterman TH, Carlsson M, Simonsson B, Bengtsson M, Nilsson K . T-cell activation and subset patterns are altered in B-CLL and correlate with the stage of the disease. Blood 1989; 74: 786–792.

    CAS  PubMed  Google Scholar 

  19. Brandl M, Grosse-Hovest L, Holler E, Kolb HJ, Jung G . Bispecific antibody fragments with CD20 × CD28 specificity allow effective autologous and allogeneic T-cell activation against malignant cells in peripheral blood and bone marrow cultures from patients with B-cell lineage leukemia and lymphoma. Exp Hematol 1999; 27: 1264–1270.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr Helmut R Salih, Medical Department II, University of Tübingen, for providing blood samples of CLL patients. This work was supported by grants from the Stiftung Deutsche Krebshilfe (Nr. 106769) and from the Deutsche Forschungsgemeinschaft (SFB773).

Author information

Author notes

  1. T Otz and L Große-Hovest: Both these authors contributed equally to this work.

Authors and Affiliations

  1. Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany

    T Otz, L Große-Hovest, M Hofmann, H-G Rammensee & G Jung

Authors
  1. T Otz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. L Große-Hovest
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M Hofmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H-G Rammensee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. G Jung
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G Jung.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Otz, T., Große-Hovest, L., Hofmann, M. et al. A bispecific single-chain antibody that mediates target cell-restricted, supra-agonistic CD28 stimulation and killing of lymphoma cells. Leukemia 23, 71–77 (2009). https://doi.org/10.1038/leu.2008.271

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/leu.2008.271

Keywords

This article is cited by

Search

Advanced search

Quick links