Chronic retroviral infection of mice promotes tumor development, but CD137 agonist therapy restores effective tumor immune surveillance
- 76 Downloads
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.
KeywordsCostimulatory 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
Fixable viability dye
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.
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.
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.
- 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.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.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
- 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.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
- 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.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
- 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
- 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.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.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.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.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.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.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
- 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.Old LJ, Boyse EA, Stockert E (1965) The G (Gross) leukemia antigen. Cancer Res 25(6):813–819Google Scholar
- 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
- 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
- 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
- 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