Chronic retroviral infection of mice promotes tumor development, but CD137 agonist therapy restores effective tumor immune surveillance

  • Anna MalyshkinaEmail author
  • Elisabeth Littwitz-Salomon
  • Kathrin Sutter
  • Jean Alexander Ross
  • Annette Paschen
  • Sonja Windmann
  • Simone Schimmer
  • Ulf Dittmer
Original Article


T cell responses are crucial for anti-tumor immunity. In chronic viral infections, anti-tumor T cell responses can be compromised due to various immunological mechanisms, including T cell exhaustion. To study mechanisms of anti-tumor immunity during a chronic viral infection, we made use of the well-established Friend virus (FV) mouse model. Chronically FV-infected mice are impaired in their ability to reject FBL-3 cells—a virus-induced tumor cell line of C57BL/6 origin. Here we aimed to explore therapeutic strategies to overcome the influence of T cell exhaustion during chronic viral infection, and reactivate effector CD8+ and CD4+ T cells to eliminate tumor cells. For T cell stimulation, agonistic antibodies against the tumor necrosis factor receptor (TNFR) superfamily members CD137 and CD134 were used, because they were reported to augment the cytotoxic program of T cells. αCD137 agonistic therapy, but not αCD134 agonistic therapy, resulted in FBL-3 tumor elimination in chronically FV-infected mice. CD137 stimulation significantly enhanced the cytotoxic activity of both CD4+ and CD8+ T cells, which were both required for efficient tumor control. Our study suggests that agonistic antibodies to CD137 can efficiently enhance anti-tumor immunity even in the setting of chronic viral infection, which might have promising therapeutic applications.


Costimulatory molecule Anti-tumor immunity Agonistic antibody Friend retrovirus Effector T cells 



Chemically induced tumor cell line




Friend virus-induced tumor cell line


Friend murine leukemia virus


Friend virus


Fixable viability dye


Granzyme B




Author contributions

Anna Malyshkina and Ulf Dittmer conceived the presented study and wrote the manuscript. Anna Malyshkina and Elisabeth Littwitz-Salomon carried out the experiments and analyzed data. Anna Malyshkina, Elisabeth Littwitz-Salomon, Sonja Windmann, Jean Alexander Ross, and Simone Schimmer were involved in the sample preparation. Kathrin Sutter and Annette Paschen contributed to the interpretation of the results. Jean Alexander Ross assisted with the design of the figures. All authors discussed the results, provided critical feedback and contributed to the final manuscript.


This work was supported by the Wilhelm Sander-Stiftung grant No 2014.091.1.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. Animal experiments were performed in strict accordance with the German regulations of the Society for Laboratory Animal Science (GV-SOLAS) and the European Health Law of the Federation of Laboratory Animal Science Associations (FELASA). The protocol was approved by the North Rhine-Westphalia State Agency for Nature, Environment and Consumer Protection (LANUV) (Permit number: G 1518/15). All efforts were made to minimize suffering.

Animal source

Female C57BL/6 mice between 6 and 10 weeks old were purchased from Envigo, Germany.

Cell line authentication

The FBL-3 cell line is a Friend virus-induced leukemia cell line, generated in a C57BL/6 mouse. The EL-4 cell line is a chemically induced lymphoma cell line, generated in a C57BL/6 mouse by 9,10-dimethyl-1,2-benzanthracene. Both cell lines were a gift from Kim J. Hasenkrug (Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA). They were expanded, aliquoted, and frozen for further use. Once in culture, cells were not continuously passaged. The identity of the cell lines was confirmed by biological assays. Before the experiments, cell lines were tested in naïve C57BL/6 mice: FBL-3 cells inoculated into the right flank of the mice were rejected, whereas EL-4 cells were not.

Supplementary material

262_2019_2300_MOESM1_ESM.pdf (409 kb)
Supplementary material 1 (PDF 408 KB)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Anna Malyshkina
    • 1
    Email author return OK on get
  • Elisabeth Littwitz-Salomon
    • 1
    • 2
  • Kathrin Sutter
    • 1
  • Jean Alexander Ross
    • 1
  • Annette Paschen
    • 3
  • Sonja Windmann
    • 1
  • Simone Schimmer
    • 1
  • Ulf Dittmer
    • 1
  1. 1.Institute for Virology, University Hospital EssenUniversity of Duisburg-EssenEssenGermany
  2. 2.School of Biochemistry and Immunology, Trinity Biomedical Sciences InstituteTrinity CollegeDublinIreland
  3. 3.Department of Dermatology, University Hospital EssenUniversity of Duisburg-EssenEssenGermany

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