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Antibody-based delivery of tumor necrosis factor (L19-TNFα) and interleukin-2 (L19-IL2) to tumor-associated blood vessels has potent immunological and anticancer activity in the syngeneic J558L BALB/c myeloma model

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

Abstract

Purpose

To analyze the impact of TNFα or IL2 on human lymphocytes in vitro and the anti-tumor and immune-modifying effects of L19-IL2 and L19-TNFα on subcutaneously growing J558L myeloma in immunocompetent mice.

Methods

PBMCs from three healthy volunteers were incubated with IL2, TNFα, or with IL2 plus addition of TNFα (final 20 h). BALB/c J558L mice with subcutaneous tumors were treated with intravenous L19-TNFα plus L19-IL2, or controls. Tumor growth and intra- and peri-tumoral tissues were analyzed for micro-vessel density, necrosis, immune cell composition, and PD1 or PD-L1 expressing cells.

Results

Exposure of PBMC in vitro to IL2, TNFα, or to IL2 over 3 and 5 days plus TNFα for the final 20 h resulted in an approximately 50 and 75% reduction of the CD25low effector cell/CD25high Treg cell ratio, respectively, compared to medium control. IL2 or TNFα increased the proportion of CD4− CD25low effector lymphocytes while reducing the proportion of CD4+ CD25low Teff cells. In the J558L myeloma model, tumor eradication was observed in 58, 42, 25, and 0% of mice treated with L19-TNFα plus L19-IL2, L19-TNFα, L19-IL2, and PBS, respectively. L19-TNFα/L19-IL2 combination caused tumor necrosis, capillary density doubling, peri-tumoral T cell and PD1+ T cell reduction (− 50%), and an increase in PD-L1+ myeloma cells.

Conclusion

IL2, TNFα, or IL2 plus TNFα (final 20 h) increased the proportion of CD4− CD25low effector lymphocytes possibly indicating immune activation. L19-TNFα/L19-IL2 combination therapy eradicated tumors in J558L myeloma BALB/c mice likely via TNFα-induced tumor necrosis and L19-TNFα/L19-IL2-mediated local cellular immune reactions.

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Acknowledgements

We thank Paulina Kuczma, Ines Puschendorf, Katja Dörfel, and Edda von der Wall for thoroughly conducting the animal studies and for the immunohistological stainings.

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Authors and Affiliations

  1. Division of Hematology and Oncology, Campus Benjamin Franklin, Department of Medicine, Charité-Universitätsmedizin Berlin, Hindenburgdamm 30, 12203, Berlin, Germany

    Hans D. Menssen

  2. Division of Oncology and Hematology, Campus Mitte, Department of Medicine, Charité-Universitätsmedizin Berlin, Charité-Platz 1, 10117, Berlin, Germany

    Ulf Harnack

  3. Division of Gastroenterology, Infectious Diseases and Rheumatology, Medical Department, Campus Benjamin Franklin, Charité-Universitätsmedizin Berlin, Hindenburgdamm 30, 12203, Berlin, Germany

    Ulrike Erben

  4. Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, ETH Zurich, Wolfgang-Pauli-Str. 10, 8093, Zurich, Switzerland

    Dario Neri

  5. Department of Pathology at Campus Benjamin Franklin, Campus Mitte, Institute of Pathology, Charité-Universitätsmedizin Berlin, Virchowweg 15, 10117, Berlin, Germany

    Burkhard Hirsch

  6. Department of Medicine, Campus Mitte, Institute of Pathology, Charité-Universitätsmedizin Berlin, Virchowweg 15, 10117, Berlin, Germany

    Burkhard Hirsch

  7. Pathodiagnostik Berlin, Komturstrasse 58-62, 12099, Berlin, Germany

    Horst Dürkop

Authors
  1. Hans D. Menssen
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  2. Ulf Harnack
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  3. Ulrike Erben
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  4. Dario Neri
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  5. Burkhard Hirsch
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Contributions

HDM, DN, and HD designed the overall study concept and the animal studies. UH designed and performed the in vitro studies on human PBMCs. UE and BH performed the immunohistologic studies. HDM, UH, and HD wrote the manuscript with the help of UE and BH.

Corresponding author

Correspondence to Hans D. Menssen.

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The authors declare no conflict of interest.

Statement of human rights

All procedures performed in studies involving human participants (blood samples obtained from human volunteers) were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Statement of animal welfare

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Menssen, H.D., Harnack, U., Erben, U. et al. Antibody-based delivery of tumor necrosis factor (L19-TNFα) and interleukin-2 (L19-IL2) to tumor-associated blood vessels has potent immunological and anticancer activity in the syngeneic J558L BALB/c myeloma model. J Cancer Res Clin Oncol 144, 499–507 (2018). https://doi.org/10.1007/s00432-017-2564-6

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  • DOI: https://doi.org/10.1007/s00432-017-2564-6

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