Abstract
The role of the immune response to oncolytic Herpes simplex viral (oHSV) therapy for glioblastoma is controversial because it might enhance or inhibit efficacy. We found that within hours of oHSV infection of glioblastomas in mice, activated natural killer (NK) cells are recruited to the site of infection. This response substantially diminished the efficacy of glioblastoma virotherapy. oHSV-activated NK cells coordinated macrophage and microglia activation within tumors. In vitro, human NK cells preferentially lysed oHSV-infected human glioblastoma cell lines. This enhanced killing depended on the NK cell natural cytotoxicity receptors (NCRs) NKp30 and NKp46, whose ligands are upregulated in oHSV-infected glioblastoma cells. We found that HSV titers and oHSV efficacy are increased in Ncr1−/− mice and a Ncr1−/− NK cell adoptive transfer model of glioma, respectively. These results demonstrate that glioblastoma virotherapy is limited partially by an antiviral NK cell response involving specific NCRs, uncovering new potential targets to enhance cancer virotherapy.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Change history
07 October 2013
In the version of this article initially published, the Online Methods incorrectly stated that mouse NK cells were treated with a blocking antibody to mouse NKp46 called BAB281. The correct antibody used to treat the cells was 29A1.4. The error has been corrected in the HTML and PDF versions of the article.
References
Wen, P.Y. & Kesari, S. Malignant gliomas in adults. N. Engl. J. Med. 359, 492–507 (2008).
Chiocca, E.A. Oncolytic viruses. Nat. Rev. Cancer 2, 938–950 (2002).
Markert, J.M. et al. Phase Ib trial of mutant herpes simplex virus G207 inoculated pre-and post-tumor resection for recurrent GBM. Mol. Ther. 17, 199–207 (2009).
Chiocca, E.A. The host response to cancer virotherapy. Curr. Opin. Mol. Ther. 10, 38–45 (2008).
Fulci, G. et al. Depletion of peripheral macrophages and brain microglia increases brain tumor titers of oncolytic viruses. Cancer Res. 67, 9398–9406 (2007).
Ikeda, K. et al. Oncolytic virus therapy of multiple tumors in the brain requires suppression of innate and elicited antiviral responses. Nat. Med. 5, 881–887 (1999).
Fulci, G. et al. Cyclophosphamide enhances glioma virotherapy by inhibiting innate immune responses. Proc. Natl. Acad. Sci. USA 103, 12873–12878 (2006).
Kurozumi, K. et al. Effect of tumor microenvironment modulation on the efficacy of oncolytic virus therapy. J. Natl. Cancer Inst. 99, 1768–1781 (2007).
Friedman, A., Tian, J.P., Fulci, G., Chiocca, E.A. & Wang, J. Glioma virotherapy: effects of innate immune suppression and increased viral replication capacity. Cancer Res. 66, 2314–2319 (2006).
Wakimoto, H., Fulci, G., Tyminski, E. & Chiocca, E.A. Altered expression of antiviral cytokine mRNAs associated with cyclophosphamide's enhancement of viral oncolysis. Gene Ther. 11, 214–223 (2004).
Altomonte, J. et al. Enhanced oncolytic potency of vesicular stomatitis virus through vector-mediated inhibition of NK and NKT cells. Cancer Gene Ther. 16, 266–278 (2009).
Todo, T., Martuza, R.L., Rabkin, S.D. & Johnson, P.A. Oncolytic herpes simplex virus vector with enhanced MHC class I presentation and tumor cell killing. Proc. Natl. Acad. Sci. USA 98, 6396–6401 (2001).
Varghese, S., Rabkin, S.D., Nielsen, P.G., Wang, W. & Martuza, R.L. Systemic oncolytic herpes virus therapy of poorly immunogenic prostate cancer metastatic to lung. Clin. Cancer Res. 12, 2919–2927 (2006).
Farrell, C.J. et al. Combination immunotherapy for tumors via sequential intratumoral injections of oncolytic herpes simplex virus 1 and immature dendritic cells. Clin. Cancer Res. 14, 7711–7716 (2008).
Hellums, E.K. et al. Increased efficacy of an interleukin-12-secreting herpes simplex virus in a syngeneic intracranial murine glioma model. Neuro-oncol. 7, 213–224 (2005).
Prestwich, R.J. et al. The case of oncolytic viruses versus the immune system: waiting on the judgment of Solomon. Hum. Gene Ther. 20, 1119–1132 (2009).
Stanford, M.M., Breitbach, C.J., Bell, J.C. & McFadden, G. Innate immunity, tumor microenvironment and oncolytic virus therapy: friends or foes? Curr. Opin. Mol. Ther. 10, 32–37 (2008).
Errington, F. et al. Reovirus activates human dendritic cells to promote innate antitumor immunity. J. Immunol. 180, 6018–6026 (2008).
Prestwich, R.J. et al. Reciprocal human dendritic cell–natural killer cell interactions induce antitumor activity following tumor cell infection by oncolytic reovirus. J. Immunol. 183, 4312–4321 (2009).
Kottke, T. et al. Use of biological therapy to enhance both virotherapy and adoptive T-cell therapy for cancer. Mol. Ther. 16, 1910–1918 (2008).
Kottke, T. et al. Improved systemic delivery of oncolytic reovirus to established tumors using preconditioning with cyclophosphamide-mediated treg modulation and interleukin-2. Clin. Cancer Res. 15, 561–569 (2009).
Derubertis, B.G. et al. Cytokine-secreting herpes viral mutants effectively treat tumor in a murine metastatic colorectal liver model by oncolytic and T-cell–dependent mechanisms. Cancer Gene Ther. 14, 590–597 (2007).
Chisholm, S.E., Howard, K., Gómez, M.V. & Reyburn, H.T. Expression of ICP0 is sufficient to trigger natural killer cell recognition of herpes simplex virus-infected cells by natural cytotoxicity receptors. J. Infect. Dis. 195, 1160–1168 (2007).
Kambara, H., Okano, H., Chiocca, E.A. & Saeki, Y. An oncolytic HSV-1 mutant expressing ICP34.5 under control of a nestin promoter increases survival of animals even when symptomatic from a brain tumor. Cancer Res. 65, 2832–2839 (2005).
Aghi, M., Visted, T., Depinho, R.A. & Chiocca, E.A. Oncolytic herpes virus with defective ICP6 specifically replicates in quiescent cells with homozygous genetic mutations in p16. Oncogene 27, 4249–4254 (2008).
Hayakawa, Y. & Smyth, M.J. CD27 dissects mature NK cells into two subsets with distinct responsiveness and migratory capacity. J. Immunol. 176, 1517–1524 (2006).
Dalton, D.K. et al. Multiple defects of immune cell function in mice with disrupted interferon-γ genes. Science 259, 1739–1742 (1993).
Yoshino, H. et al. Natural killer cell depletion by anti-asialo GM1 antiserum treatment enhances human hematopoietic stem cell engraftment in NOD/Shi-scid mice. Bone Marrow Transplant. 26, 1211–1216 (2000).
Randolph, G.J., Jakubzick, C. & Qu, C. Antigen presentation by monocytes and monocyte-derived cells. Curr. Opin. Immunol. 20, 52–60 (2008).
Savarin, C. & Bergmann, C.C. Neuroimmunology of central nervous system viral infections: the cells, molecules and mechanisms involved. Curr. Opin. Pharmacol. 8, 472–479 (2008).
Weiner, N.E. et al. A syngeneic mouse glioma model for study of glioblastoma therapy. J. Neuropathol. Exp. Neurol. 58, 54–60 (1999).
Taylor, M.A., Ward, B., Schatzle, J.D. & Bennett, M. Perforin- and Fas-dependent mechanisms of natural killer cell–mediated rejection of incompatible bone marrow cell grafts. Eur. J. Immunol. 32, 793–799 (2002).
Trotta, R. et al. Dependence of both spontaneous and antibody-dependent, granule exocytosis-mediated NK cell cytotoxicity on extracellular signal-regulated kinases. J. Immunol. 161, 6648–6656 (1998).
Castriconi, R. et al. NK cells recognize and kill human glioblastoma cells with stem cell–like properties. J. Immunol. 182, 3530–3539 (2009).
Wu, A. et al. Expression of MHC I and NK ligands on human CD133+ glioma cells: possible targets of immunotherapy. J. Neurooncol. 83, 121–131 (2007).
Brandt, C.S. et al. The B7 family member B7–H6 is a tumor cell ligand for the activating natural killer cell receptor NKp30 in humans. J. Exp. Med. 206, 1495–1503 (2009).
Orange, J.S. Human natural killer cell deficiencies. Curr. Opin. Allergy Clin. Immunol. 6, 399–409 (2006).
Altomonte, J. et al. Exponential enhancement of oncolytic vesicular stomatitis virus potency by vector-mediated suppression of inflammatory responses in vivo. Mol. Ther. 16, 146–153 (2008).
Galivo, F. et al. Interference of CD40L-mediated tumor immunotherapy by oncolytic vesicular stomatitis virus. Hum. Gene Ther. 21, 439–450 (2010).
Galivo, F. et al. Single-cycle viral gene expression, rather than progressive replication and oncolysis, is required for VSV therapy of B16 melanoma. Gene Ther. 17, 158–170 (2010).
Marques, C.P., Hu, S., Sheng, W. & Lokensgard, J.R. Microglial cells initiate vigorous yet non-protective immune responses during HSV-1 brain infection. Virus Res. 121, 1–10 (2006).
Yu, J. et al. CD94 surface density identifies a functional intermediary between the CD56bright and CD56dim human NK cell subsets. Blood 115, 274–281 (2010).
Lundberg, P. et al. A locus on mouse chromosome 6 that determines resistance to herpes simplex virus also influences reactivation, while an unlinked locus augments resistance of female mice. J. Virol. 77, 11661–11673 (2003).
Arnon, T.I., Markel, G. & Mandelboim, O. Tumor and viral recognition by natural killer cells receptors. Semin. Cancer Biol. 16, 348–358 (2006).
Bloushtain, N. et al. Membrane-associated heparan sulfate proteoglycans are involved in the recognition of cellular targets by NKp30 and NKp46. J. Immunol. 173, 2392–2401 (2004).
Ferlazzo, G. et al. Human dendritic cells activate resting natural killer (NK) cells and are recognized via the NKp30 receptor by activated NK cells. J. Exp. Med. 195, 343–351 (2002).
Arnon, T.I. et al. Inhibition of the NKp30 activating receptor by pp65 of human cytomegalovirus. Nat. Immunol. 6, 515–523 (2005).
Degli-Esposti, M.A. & Smyth, M.J. Close encounters of different kinds: dendritic cells and NK cells take centre stage. Nat. Rev. Immunol. 5, 112–124 (2005).
Pogge von Strandmann, E. et al. Human leukocyte antigen-B–associated transcript 3 is released from tumor cells and engages the NKp30 receptor on natural killer cells. Immunity 27, 965–974 (2007).
Sivori, S. et al. NKp46 is the major triggering receptor involved in the natural cytotoxicity of fresh or cultured human NK cells. Correlation between surface density of NKp46 and natural cytotoxicity against autologous, allogeneic or xenogeneic target cells. Eur. J. Immunol. 29, 1656–1666 (1999).
Giannini, C. et al. Patient tumor EGFR and PDGFRA gene amplifications retained in an invasive intracranial xenograft model of glioblastoma multiforme. Neuro-oncol. 7, 164–176 (2005).
Reilly, K.M., Loisel, D.A., Bronson, R.T., McLaughlin, M.E. & Jacks, T. Nf1;Trp53 mutant mice develop glioblastoma with evidence of strain-specific effects. Nat. Genet. 26, 109–113 (2000).
Shultz, L.D. et al. Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2Rγ null mice engrafted with mobilized human hemopoietic stem cells. J. Immunol. 174, 6477–6489 (2005).
Yu, J. et al. NKp46 identifies an NKT cell subset susceptible to leukemic transformation in mouse and human. J. Clin. Invest. 121, 1456–1470 (2011).
Ghiasi, H., Cai, S., Perng, G.C., Nesburn, A.B. & Wechsler, S.L. The role of natural killer cells in protection of mice against death and corneal scarring following ocular HSV-1 infection. Antiviral Res. 45, 33–45 (2000).
Gazit, R. et al. Lethal influenza infection in the absence of the natural killer cell receptor gene Ncr1. Nat. Immunol. 7, 517–523 (2006).
Marques, C.P. et al. Prolonged microglial cell activation and lymphocyte infiltration following experimental herpes encephalitis. J. Immunol. 181, 6417–6426 (2008).
Vitale, M. et al. NK-dependent DC maturation is mediated by TNFα and IFNγ released upon engagement of the NKp30 triggering receptor. Blood 106, 566–571 (2005).
Walzer, T. et al. Identification, activation, and selective in vivo ablation of mouse NK cells via NKp46. Proc. Natl. Acad. Sci. USA 104, 3384–3389 (2007).
Acknowledgements
This work was supported by US National Institutes of Health grants 7U01NS061811 (to E.A.C.), CA069246 (to E.A.C.), CA068458 (to M.A.C.), CA095426 (to M.A.C.), TL1RR025753 (to C.A.A.-B.) and CA163205 (to E.A.C., M.A.C. and B.K.). C.A.A.-B. was supported by an American Medical Association Foundation Seed Grant. This work was also supported by the Dardinger Neuro-oncology Laboratory.
Author information
Authors and Affiliations
Contributions
C.A.A.-B., J.Y., R.P., J.W., J.P., H.M., M.W., Y.W., S.H. and J.H. performed experiments. C.A.A.-B., J.Y., B.K., S.E.L., A.M., M.A.C. and E.A.C. conceived the experimental approach, directed experiments and interpreted data. S.A.F. performed statistical analysis. E.V., O.M. and A.M. provided reagents. C.A.A.-B., J.Y., M.A.C. and E.A.C. wrote the manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–6 and Supplementary Tables 1 and 2 (PDF 1170 kb)
Rights and permissions
About this article
Cite this article
Alvarez-Breckenridge, C., Yu, J., Price, R. et al. NK cells impede glioblastoma virotherapy through NKp30 and NKp46 natural cytotoxicity receptors. Nat Med 18, 1827–1834 (2012). https://doi.org/10.1038/nm.3013
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nm.3013
This article is cited by
-
NK cells as powerful therapeutic tool in cancer immunotherapy
Cellular Oncology (2024)
-
Natural killer cell-related gene signature predicts malignancy of glioma and the survival of patients
BMC Cancer (2022)
-
Cutting both ways: the innate immune response to oncolytic virotherapy
Cancer Gene Therapy (2022)
-
Natural killer cell homing and trafficking in tissues and tumors: from biology to application
Signal Transduction and Targeted Therapy (2022)
-
Treat and repeat: oncolytic virus therapy for brain cancer
Nature Medicine (2022)