Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Altered expression of antiviral cytokine mRNAs associated with cyclophosphamide's enhancement of viral oncolysis

Gene Therapy volume 11, pages 214–223 (2004)Cite this article

Abstract

Oncolytic viruses (OVs) are being used as anticancer agents in preclinical and clinical trials. Propagation of OVs inside infected tumors is critical to their efficacy and is mediated by the productive generation of progeny OVs within infected tumor cells. In turn, this progeny can spread the infection to other tumor cells in successive rounds of oncolysis. Previously, we had found that, in rats, cyclophosphamide (CPA) pretreatment increased infection of brain tumors by an intra-arterially administered herpes simplex virus type 1 OV, because it inhibited activation of complement responses, mediated by innate IgM. We also have previously shown that other pharmacologic inhibitors of complement, such as cobra venom factor (CVF), allowed for increased infection. However, in these studies, further inhibition of complement responses by CVF did not result in additional infection of brain tumor cells or in propagation of OV to surrounding tumor cells. In this study, we sought to determine if CPA did lead to increased infection/propagation from initially infected tumor cells. Unlike our results with CVF, we find that CPA administration does result in a time-dependent increase in infection of tumor cells, suggestive of increased propagation, in both syngeneic and athymic models of brain tumors. This increase was due to increased survival of OV within infected tumors and brain surrounding tumors. CPA's effect was not due to a direct enhancement of viral replication in tumor cells, rather was associated with its immunosuppressive effects. RT-PCR analysis revealed that CPA administration resulted in impaired mRNA production by peripheral blood mononuclear cells (PBMCs) of several cytokines (interferons α/β, interferon γ, TNFα, IL-15, and IL-18) with anti-HSV function. These findings suggest that the CPA-mediated facilitation of OV intraneoplastic propagation is associated with a general decrease of antiviral cytokines mRNAs in PBMCs. These findings not only suggest a potential benefit for the addition of transient immunosuppression in clinical applications of oncolytic HSV therapy, but also suggest that innate immunomodulatory pathways may be amenable to manipulation, in order to increase OV propagation and survival within infected tumors.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Chiocca EA, Smith ER . Oncolytic viruses as novel anticancer agents: turning one scourge against another (In Process Citation). Expert Opin Investig Drugs 2000; 9: 311–327 (MEDLINE record in process).

    Article  PubMed  Google Scholar 

  2. Kirn D, Martuza RL, Zwiebel J . Replication-selective virotherapy for cancer: biological principles, risk management and future directions. Nat Med 2001; 7: 781–787.

    Article  CAS  PubMed  Google Scholar 

  3. Chiocca EA . Oncolytic viruses. Nat Rev Cancer 2002; 2: 938–950.

    Article  PubMed  Google Scholar 

  4. Markert JM et al. Conditionally replicating herpes simplex virus mutant, G207 for the treatment of malignant glioma: results of a phase I trial (see comments). Gene Therapy 2000; 7: 867–874.

    Article  CAS  PubMed  Google Scholar 

  5. Rampling R et al. Toxicity evaluation of replication-competent herpes simplex virus (ICP 34.5 null mutant 1716) in patients with recurrent malignant glioma. Gene Therapy 2000; 7: 859–866.

    Article  CAS  PubMed  Google Scholar 

  6. Khuri FR et al. A controlled trial of intratumoral ONYX-015, a selectively-replicating adenovirus, in combination with cisplatin and 5-fluorouracil in patients with recurrent head and neck cancer (see comments). Nat Med 2000; 6: 879–885.

    Article  CAS  PubMed  Google Scholar 

  7. Ichikawa T, Chiocca EA . Comparative analyses of transgene delivery and expression in tumors inoculated with a replication-conditional or -defective viral vector. Cancer Res 2001; 61: 5336–5339.

    CAS  PubMed  Google Scholar 

  8. Wakimoto H, Johnson PR, Knipe DM, Chiocca EA . Effects of innate immunity on herpes simplex virus and its ability to kill tumor cells. Gene Therapy 2003; 10: 983–990.

    Article  CAS  PubMed  Google Scholar 

  9. Mitchell BM, Stevens JG . Neuroinvasive properties of herpes simplex virus type 1 glycoprotein variants are controlled by the immune response. J Immunol 1996; 156: 246–255.

    CAS  PubMed  Google Scholar 

  10. Fawaz LM, Sharif-Askari E, Menezes J . Up-regulation of NK cytotoxic activity via IL-15 induction by different viruses: a comparative study. J Immunol 1999; 163: 4473–4480.

    CAS  PubMed  Google Scholar 

  11. Kodukula P et al. Macrophage control of herpes simplex virus type 1 replication in the peripheral nervous system. J Immunol 1999; 162: 2895–2905.

    CAS  PubMed  Google Scholar 

  12. Feduchi E, Alonso MA, Carrasco L . Human gamma interferon and tumor necrosis factor exert a synergistic blockade on the replication of herpes simplex virus. J Virol 1989; 63: 1354–1359.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Goodbourn S, Didcock L, Randall RE . Interferons: cell signalling, immune modulation, antiviral response and virus countermeasures. J Gen Virol 2000; 81: 2341–2364.

    Article  CAS  PubMed  Google Scholar 

  14. Ahmad A, Sharif-Askari E, Fawaz L, Menezes J . Innate immune response of the human host to exposure with herpes simplex virus type 1: in vitro control of the virus infection by enhanced natural killer activity via interleukin-15 induction. J Virol 2000; 74: 7196–7203.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Fujioka N et al. Interleukin-18 protects mice against acute herpes simplex virus type 1 infection (viruses: a comparative study). J Virol 1999; 73: 2401–2409.

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Karupiah G et al. Inhibition of viral replication by interferon-gamma-induced nitric oxide synthase. Science 1993; 261: 1445–1448.

    Article  CAS  PubMed  Google Scholar 

  17. Croen KD . Evidence for antiviral effect of nitric oxide. Inhibition of herpes simplex virus type 1 replication. J Clin Invest 1993; 91: 2446–2452.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Leib DA et al. Specific phenotypic restoration of an attenuated virus by knockout of a host resistance gene. Proc Natl Acad Sci USA 2000; 97: 6097–6101.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Ikeda K et al. Complement depletion facilitates the infection of multiple brain tumors by an intravascular, replication-conditional herpes simplex virus mutant. J Virol 2000; 74: 4765–4775.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Ikeda K et al. Oncolytic virus therapy of multiple tumors in the brain requires suppression of innate and elicited antiviral responses. Nat Med 1999; 5: 881–887.

    Article  CAS  PubMed  Google Scholar 

  21. Wakimoto H et al. The complement response against an oncolytic virus is species-specific in its activation pathways. Mol Ther 2002; 5: 275–282.

    Article  CAS  PubMed  Google Scholar 

  22. Tzeng JJ, Barth RF, Orosz CG, James SM . Phenotype and functional activity of tumor-infiltrating lymphocytes isolated from immunogenic and nonimmunogenic rat brain tumors. Cancer Res 1991; 51: 2373–2378.

    CAS  PubMed  Google Scholar 

  23. Geraghty RJ et al. Entry of alpha herpesviruses mediated by poliovirus receptor-related protein 1 and poliovirus receptor. Science 1998; 280: 1618–1620.

    Article  CAS  PubMed  Google Scholar 

  24. Kramm CM et al. Therapeutic efficiency and safety of a second-generation replication-conditional HSV1 vector for brain tumor gene therapy. Hum Gene Ther 1997; 8: 2057–2068.

    Article  CAS  PubMed  Google Scholar 

  25. Advani SJ et al. Enhancement of replication of genetically engineered herpes simplex viruses by ionizing radiation: a new paradigm for destruction of therapeutically intractable tumors. Gene Therapy 1998; 5: 160–165.

    Article  CAS  PubMed  Google Scholar 

  26. Clarke L, Waxman DJ . Oxidative metabolism of cyclophosphamide: identification of the hepatic monooxygenase catalysts of drug activation. Cancer Res 1989; 49: 2344–2350.

    CAS  PubMed  Google Scholar 

  27. Matar P, Rozados VR, Gervasoni SI, Scharovsky GO . Th2/Th1 switch induced by a single low dose of cyclophosphamide in a rat metastatic lymphoma model. Cancer Immunol Immunother 2002; 50: 588–596.

    Article  CAS  PubMed  Google Scholar 

  28. Leib DA . Counteraction of interferon-induced antiviral responses by herpes simplex viruses. Curr Top Microbiol Immunol 2002; 269: 171–185.

    CAS  PubMed  Google Scholar 

  29. Leib DA et al. Interferons regulate the phenotype of wild-type and mutant herpes simplex viruses in vivo. J Exp Med 1999; 189: 663–672.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Zamanian-Daryoush M, Mogensen TH, DiDonato JA, Williams BR . NF-kappaB activation by double-stranded-RNA-activated protein kinase (PKR) is mediated through NF-kappaB-inducing kinase and IkappaB kinase. Mol Cell Biol 2000; 20: 1278–1290.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Deb A et al. RNA-dependent protein kinase PKR is required for activation of NF-kappa B by IFN-gamma in a STAT1-independent pathway. J Immunol 2001; 166: 6170–6180.

    Article  CAS  PubMed  Google Scholar 

  32. Uetani K et al. Central role of double-stranded RNA-activated protein kinase in microbial induction of nitric oxide synthase. J Immunol 2000; 165: 988–996.

    Article  CAS  PubMed  Google Scholar 

  33. Geiger KD et al. Interferon-gamma protects against herpes simplex virus type 1-mediated neuronal death. Virology 1997; 238: 189–197.

    Article  CAS  PubMed  Google Scholar 

  34. Halford WP, Schaffer PA . Optimized viral dose and transient immunosuppression enable herpes simplex virus ICP0-null mutants to establish wild-type levels of latency in vivo. J Virol 2000; 74: 5957–5967.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Hickman-Davis JM, Lindsey JR, Matalon S . Cyclophosphamide decreases nitrotyrosine formation and inhibits nitric oxide production by alveolar macrophages in mycoplasmosis. Infect Immun 2001; 69: 6401–6410.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Wei MX et al. Diffusible cytotoxic metabolites contribute to the in vitro bystander effect associated with the cyclophosphamide/cytochrome P450 2B1 cancer gene therapy paradigm. Clin Cancer Res 1995; 1: 1171–1177.

    CAS  PubMed  Google Scholar 

  37. Goldstein DJ, Weller SK . Herpes simplex virus type 1-induced ribonucleotide reductase activity is dispensable for virus growth and DNA synthesis: isolation and characterization of an ICP6 lacZ insertion mutant. J Virol 1988; 62: 196–205.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Goldstein DJ, Weller SK . Factor(s) present in herpes simplex virus type 1-infected cells can compensate for the loss of the large subunit of the viral ribonucleotide reductase: characterization of an ICP6 deletion mutant. Virology 1988; 166: 41–51.

    Article  CAS  PubMed  Google Scholar 

  39. Goldstein DJ, Weller SK . An ICP6::lacZ insertional mutagen is used to demonstrate that the UL52 gene of herpes simplex virus type 1 is required for virus growth and DNA synthesis. J Virol 1988; 62: 2970–2977.

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Boviatsis EJ et al. Gene transfer into experimental brain tumors mediated by adenovirus, herpes simplex virus, and retrovirus vectors. Hum Gene Ther 1994; 5: 183–191.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Y Saeki and other members of the EAC laboratory for their input in the described experiments. We acknowledge J Basilion and M Pasternack for useful discussions. We dedicate this work to the memory of Dr Keiro Ikeda (MGH and Keio University). This work was supported by a NIH research Grant (CA 69246), the Berkowitz-Knott Fund for Brain Tumor Research, and a grant from the Uehara Memorial Foundation (Tokyo, Japan).

Author information

Authors and Affiliations

  1. Molecular Neuro-Oncology Laboratories, Neurosurgery Service, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA

    H Wakimoto, G Fulci, E Tyminski & E Antonio Chiocca

Authors
  1. H Wakimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G Fulci
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. E Tyminski
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. E Antonio Chiocca
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wakimoto, H., Fulci, G., Tyminski, E. et al. Altered expression of antiviral cytokine mRNAs associated with cyclophosphamide's enhancement of viral oncolysis. Gene Ther 11, 214–223 (2004). https://doi.org/10.1038/sj.gt.3302143

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3302143

Keywords

This article is cited by

Search

Advanced search

Quick links