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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
  • 77 Downloads

Abstract

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.

Keywords

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

Abbreviations

EL-4

Chemically induced tumor cell line

Eomes

Eomesodermin

FBL-3

Friend virus-induced tumor cell line

F-MuLV

Friend murine leukemia virus

FV

Friend virus

FVD

Fixable viability dye

GzmB

Granzyme B

Tet

Tetramer

Notes

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.

Funding

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)

References

  1. 1.
    Haabeth OA, Tveita AA, Fauskanger M, Schjesvold F, Lorvik KB, Hofgaard PO, Omholt H, Munthe LA, Dembic Z, Corthay A, Bogen B (2014) How do CD4(+) T cells detect and eliminate tumor cells that either lack or express MHC class II molecules? Front Immunol 5:174.  https://doi.org/10.3389/fimmu.2014.00174 CrossRefGoogle Scholar
  2. 2.
    Quezada SA, Simpson TR, Peggs KS, Merghoub T, Vider J, Fan X, Blasberg R, Yagita H, Muranski P, Antony PA, Restifo NP, Allison JP (2010) Tumor-reactive CD4(+) T cells develop cytotoxic activity and eradicate large established melanoma after transfer into lymphopenic hosts. J Exp Med 207(3):637–650.  https://doi.org/10.1084/jem.20091918 CrossRefGoogle Scholar
  3. 3.
    Akhmetzyanova I, Zelinskyy G, Schimmer S, Brandau S, Altenhoff P, Sparwasser T, Dittmer U (2013) Tumor-specific CD4 + T cells develop cytotoxic activity and eliminate virus-induced tumor cells in the absence of regulatory T cells. Cancer Immunol Immunother 62(2):257–271.  https://doi.org/10.1007/s00262-012-1329-y CrossRefGoogle Scholar
  4. 4.
    Zelinskyy G, Werner T, Dittmer U (2013) Natural regulatory T cells inhibit production of cytotoxic molecules in CD8(+) T cells during low-level Friend retrovirus infection. Retrovirology 10:109.  https://doi.org/10.1186/1742-4690-10-109 CrossRefGoogle Scholar
  5. 5.
    Viguier M, Lemaitre F, Verola O, Cho MS, Gorochov G, Dubertret L, Bachelez H, Kourilsky P, Ferradini L (2004) Foxp3 expressing CD4 + CD25(high) regulatory T cells are overrepresented in human metastatic melanoma lymph nodes and inhibit the function of infiltrating T cells. J Immunol 173(2):1444–1453CrossRefGoogle Scholar
  6. 6.
    Rodger AJ, Lodwick R, Schechter M, Deeks S, Amin J, Gilson R, Paredes R, Bakowska E, Engsig FN, Phillips A, Insight Smart ESG (2013) Mortality in well controlled HIV in the continuous antiretroviral therapy arms of the SMART and ESPRIT trials compared with the general population. AIDS 27(6):973–979.  https://doi.org/10.1097/QAD.0b013e32835cae9c CrossRefGoogle Scholar
  7. 7.
    Puoti M, Bruno R, Soriano V, Donato F, Gaeta GB, Quinzan GP, Precone D, Gelatti U, Asensi V, Vaccher E, Group HHCI-S (2004) Hepatocellular carcinoma in HIV-infected patients: epidemiological features, clinical presentation and outcome. AIDS 18(17):2285–2293CrossRefGoogle Scholar
  8. 8.
    Rubinstein PG, Aboulafia DM, Zloza A (2014) Malignancies in HIV/AIDS: from epidemiology to therapeutic challenges. AIDS 28(4):453–465.  https://doi.org/10.1097/QAD.0000000000000071 CrossRefGoogle Scholar
  9. 9.
    Iwashiro M, Messer RJ, Peterson KE, Stromnes IM, Sugie T, Hasenkrug KJ (2001) Immunosuppression by CD4 + regulatory T cells induced by chronic retroviral infection. Proc Natl Acad Sci U S A 98(16):9226–9230.  https://doi.org/10.1073/pnas.151174198 CrossRefGoogle Scholar
  10. 10.
    Akhmetzyanova I, Zelinskyy G, Littwitz-Salomon E, Malyshkina A, Dietze KK, Streeck H, Brandau S, Dittmer U (2016) CD137 agonist therapy can reprogram regulatory T cells into cytotoxic CD4+ T cells with antitumor activity. J Immunol 196(1):484–492.  https://doi.org/10.4049/jimmunol.1403039 CrossRefGoogle Scholar
  11. 11.
    Dittmer U, He H, Messer RJ, Schimmer S, Olbrich AR, Ohlen C, Greenberg PD, Stromnes IM, Iwashiro M, Sakaguchi S, Evans LH, Peterson KE, Yang G, Hasenkrug KJ (2004) Functional impairment of CD8(+) T cells by regulatory T cells during persistent retroviral infection. Immunity 20(3):293–303CrossRefGoogle Scholar
  12. 12.
    Li SY, Liu Y (2013) Immunotherapy of melanoma with the immune costimulatory monoclonal antibodies targeting CD137. Clin Pharmacol 5(Suppl 1):47–53.  https://doi.org/10.2147/CPAA.S46199 Google Scholar
  13. 13.
    Melero I, Murillo O, Dubrot J, Hervas-Stubbs S, Perez-Gracia JL (2008) Multi-layered action mechanisms of CD137 (4-1BB)-targeted immunotherapies. Trends Pharmacol Sci 29(8):383–390.  https://doi.org/10.1016/j.tips.2008.05.005 CrossRefGoogle Scholar
  14. 14.
    Vinay DS, Kwon BS (2012) Immunotherapy of cancer with 4-1BB. Mol Cancer Ther 11(5):1062–1070.  https://doi.org/10.1158/1535-7163.MCT-11-0677 CrossRefGoogle Scholar
  15. 15.
    Melero I, Shuford WW, Newby SA, Aruffo A, Ledbetter JA, Hellstrom KE, Mittler RS, Chen L (1997) Monoclonal antibodies against the 4-1BB T-cell activation molecule eradicate established tumors. Nat Med 3(6):682–685CrossRefGoogle Scholar
  16. 16.
    Menk AV, Scharping NE, Rivadeneira DB, Calderon MJ, Watson MJ, Dunstane D, Watkins SC, Delgoffe GM (2018) 4-1BB costimulation induces T cell mitochondrial function and biogenesis enabling cancer immunotherapeutic responses. J Exp Med 215(4):1091–1100.  https://doi.org/10.1084/jem.20171068 CrossRefGoogle Scholar
  17. 17.
    Yonezawa A, Dutt S, Chester C, Kim J, Kohrt HE (2015) Boosting cancer immunotherapy with anti-CD137 antibody therapy. Clin Cancer Res 21(14):3113–3120.  https://doi.org/10.1158/1078-0432.CCR-15-0263 CrossRefGoogle Scholar
  18. 18.
    Ju SA, Cheon SH, Park SM, Tam NQ, Kim YM, An WG, Kim BS (2008) Eradication of established renal cell carcinoma by a combination of 5-fluorouracil and anti-4-1BB monoclonal antibody in mice. Int J Cancer 122(12):2784–2790.  https://doi.org/10.1002/ijc.23457 CrossRefGoogle Scholar
  19. 19.
    Gramaglia I, Weinberg AD, Lemon M, Croft M (1998) Ox-40 ligand: a potent costimulatory molecule for sustaining primary CD4 T cell responses. J Immunol 161(12):6510–6517Google Scholar
  20. 20.
    Gough MJ, Crittenden MR, Sarff M, Pang P, Seung SK, Vetto JT, Hu HM, Redmond WL, Holland J, Weinberg AD (2010) Adjuvant therapy with agonistic antibodies to CD134 (OX40) increases local control after surgical or radiation therapy of cancer in mice. J Immunother 33(8):798–809.  https://doi.org/10.1097/CJI.0b013e3181ee7095 CrossRefGoogle Scholar
  21. 21.
    Malyshkina A, Littwitz-Salomon E, Sutter K, Zelinskyy G, Windmann S, Schimmer S, Paschen A, Streeck H, Hasenkrug KJ, Dittmer U (2017) Fas Ligand-mediated cytotoxicity of CD4+ T cells during chronic retrovirus infection. Sci Rep 7(1):7785.  https://doi.org/10.1038/s41598-017-08578-7 CrossRefGoogle Scholar
  22. 22.
    Zelinskyy G, Dietze KK, Husecken YP, Schimmer S, Nair S, Werner T, Gibbert K, Kershaw O, Gruber AD, Sparwasser T, Dittmer U (2009) The regulatory T-cell response during acute retroviral infection is locally defined and controls the magnitude and duration of the virus-specific cytotoxic T-cell response. Blood 114(15):3199–3207.  https://doi.org/10.1182/blood-2009-03-208736 CrossRefGoogle Scholar
  23. 23.
    Iwashiro M, Kondo T, Shimizu T, Yamagishi H, Takahashi K, Matsubayashi Y, Masuda T, Otaka A, Fujii N, Ishimoto A et al (1993) Multiplicity of virus-encoded helper T-cell epitopes expressed on FBL-3 tumor cells. J Virol 67(8):4533–4542Google Scholar
  24. 24.
    Chen W, Qin H, Chesebro B, Cheever MA (1996) Identification of a gag-encoded cytotoxic T-lymphocyte epitope from FBL-3 leukemia shared by Friend, Moloney, and Rauscher murine leukemia virus-induced tumors. J Virol 70(11):7773–7782Google Scholar
  25. 25.
    Pearce EL, Mullen AC, Martins GA, Krawczyk CM, Hutchins AS, Zediak VP, Banica M, DiCioccio CB, Gross DA, Mao CA, Shen H, Cereb N, Yang SY, Lindsten T, Rossant J, Hunter CA, Reiner SL (2003) Control of effector CD8+ T cell function by the transcription factor Eomesodermin. Science 302(5647):1041–1043.  https://doi.org/10.1126/science.1090148 CrossRefGoogle Scholar
  26. 26.
    Yang Y, Xu J, Niu Y, Bromberg JS, Ding Y (2008) T-bet and eomesodermin play critical roles in directing T cell differentiation to Th1 versus Th17. J Immunol 181(12):8700–8710CrossRefGoogle Scholar
  27. 27.
    Manzke N, Akhmetzyanova I, Hasenkrug KJ, Trilling M, Zelinskyy G, Dittmer U (2013) CD4+ T cells develop antiretroviral cytotoxic activity in the absence of regulatory T cells and CD8+ T cells. J Virol 87(11):6306–6313.  https://doi.org/10.1128/JVI.00432-13 CrossRefGoogle Scholar
  28. 28.
    Dietze KK, Zelinskyy G, Liu J, Kretzmer F, Schimmer S, Dittmer U (2013) Combining regulatory T cell depletion and inhibitory receptor blockade improves reactivation of exhausted virus-specific CD8 + T cells and efficiently reduces chronic retroviral loads. PLoS Pathog 9(12):e1003798.  https://doi.org/10.1371/journal.ppat.1003798 CrossRefGoogle Scholar
  29. 29.
    Old LJ, Boyse EA, Stockert E (1965) The G (Gross) leukemia antigen. Cancer Res 25(6):813–819Google Scholar
  30. 30.
    Zelinskyy G, Robertson SJ, Schimmer S, Messer RJ, Hasenkrug KJ, Dittmer U (2005) CD8+ T-cell dysfunction due to cytolytic granule deficiency in persistent Friend retrovirus infection. J Virol 79(16):10619–10626.  https://doi.org/10.1128/JVI.79.16.10619-10626.2005 CrossRefGoogle Scholar
  31. 31.
    Kitamura N, Murata S, Ueki T, Mekata E, Reilly RT, Jaffee EM, Tani T (2009) OX40 costimulation can abrogate Foxp3 + regulatory T cell-mediated suppression of antitumor immunity. Int J Cancer 125(3):630–638.  https://doi.org/10.1002/ijc.24435 CrossRefGoogle Scholar
  32. 32.
    Makkouk A, Chester C, Kohrt HE (2016) Rationale for anti-CD137 cancer immunotherapy. Eur J Cancer 54:112–119.  https://doi.org/10.1016/j.ejca.2015.09.026 CrossRefGoogle Scholar
  33. 33.
    Weigelin B, Bolanos E, Rodriguez-Ruiz ME, Martinez-Forero I, Friedl P, Melero I (2016) Anti-CD137 monoclonal antibodies and adoptive T cell therapy: a perfect marriage? Cancer Immunol Immunother 65(5):493–497.  https://doi.org/10.1007/s00262-016-1818-5 CrossRefGoogle Scholar
  34. 34.
    Segal NH, Logan TF, Hodi FS, McDermott D, Melero I, Hamid O, Schmidt H, Robert C, Chiarion-Sileni V, Ascierto PA, Maio M, Urba WJ, Gangadhar TC, Suryawanshi S, Neely J, Jure-Kunkel M, Krishnan S, Kohrt H, Sznol M, Levy R (2017) Results from an integrated safety analysis of urelumab, an agonist anti-CD137 monoclonal antibody. Clin Cancer Res 23(8):1929–1936.  https://doi.org/10.1158/1078-0432.CCR-16-1272 CrossRefGoogle Scholar
  35. 35.
    Chester C, Sanmamed MF, Wang J, Melero I (2018) Immunotherapy targeting 4-1BB: mechanistic rationale, clinical results, and future strategies. Blood 131(1):49–57.  https://doi.org/10.1182/blood-2017-06-741041 Google Scholar
  36. 36.
    Joedicke JJ, Zelinskyy G, Dittmer U, Hasenkrug KJ (2014) CD8+ T cells are essential for controlling acute friend retrovirus infection in C57BL/6 mice. J Virol 88(9):5200–5201.  https://doi.org/10.1128/JVI.00312-14 CrossRefGoogle Scholar
  37. 37.
    Dietze KK, Zelinskyy G, Gibbert K, Schimmer S, Francois S, Myers L, Sparwasser T, Hasenkrug KJ, Dittmer U (2011) Transient depletion of regulatory T cells in transgenic mice reactivates virus-specific CD8 + T cells and reduces chronic retroviral set points. Proc Natl Acad Sci U S A 108(6):2420–2425.  https://doi.org/10.1073/pnas.1015148108 CrossRefGoogle Scholar
  38. 38.
    Nair S, Bayer W, Ploquin MJ, Kassiotis G, Hasenkrug KJ, Dittmer U (2011) Distinct roles of CD4+ T cell subpopulations in retroviral immunity: lessons from the Friend virus mouse model. Retrovirology 8:76.  https://doi.org/10.1186/1742-4690-8-76 CrossRefGoogle Scholar
  39. 39.
    Udono H, Mieno M, Shiku H, Nakayama E (1989) The roles of CD8+ and CD4+ cells in tumor rejection. Jpn J Cancer Res 80(7):649–654CrossRefGoogle Scholar
  40. 40.
    Aspeslagh S, Postel-Vinay S, Rusakiewicz S, Soria JC, Zitvogel L, Marabelle A (2016) Rationale for anti-OX40 cancer immunotherapy. Eur J Cancer 52:50–66.  https://doi.org/10.1016/j.ejca.2015.08.021 CrossRefGoogle Scholar
  41. 41.
    Lai C, August S, Albibas A, Behar R, Cho SY, Polak ME, Theaker J, MacLeod AS, French RR, Glennie MJ, Al-Shamkhani A, Healy E (2016) OX40 + regulatory T cells in cutaneous squamous cell carcinoma suppress effector T-cell responses and associate with metastatic potential. Clin Cancer Res 22(16):4236–4248.  https://doi.org/10.1158/1078-0432.CCR-15-2614 CrossRefGoogle Scholar

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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