Skip to main content

Advertisement

SpringerLink
Log in

The detection of CMV pp65 and IE1 in glioblastoma multiforme

  • Laboratory Investigation - Human/Animal Tissue
  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

Glioblastoma multiforme (GBM) is a highly lethal brain tumor affecting children and adults, with the majority of affected individuals dying from their disease by 2 years following diagnosis. Other groups have reported the association of cytomegalovirus (CMV) with GBM, and we sought to confirm these findings in a large series of patients with primary GBM from our institution. Immunohistochemical analysis of paraffin embedded tissue sections was performed on 49 newly diagnosed GBM tumors, the largest series reported to date. We confirmed the presence of CMV pp65 on 25/49 (51%) and of IE1 on 8/49 (16%) of these tumors. While pp65 and IE1 are generally found in the nucleus of cells that are permissibly infected by CMV, GBM in this series had mostly cytoplasmic staining, with only 16% having nuclear staining for one or both of these antigens. We infected GBM cell lines with a laboratory strain of CMV, and found that most of the staining was cytoplasmic, with some perinuclear localization of IE1. To test the potential for CMV infected GBM cells to be recognized by CMV pp65 and IE1 specific cytotoxic T lymphocytes (CTL), we used CMV infected GBM cell lines in cytotoxicity assays with human leukocyte antigen partially matched CMV CTL. Lysis of CMV infected GBM tumor cells was accentuated by pre-treating these cell lines with either the demethylating agent decitabine or interferon-γ, both of which were shown to increase MHC Class I and II expression on tumor cells in vitro. These studies confirm the presence of CMV pp65 or IE1 on approximately half of GBM, with the possibility that CMV positive tumor cells can be recognized by CMV pp65/IE1 specific T cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Davis FG et al (2001) Prevalence estimates for primary brain tumors in the United States by behavior and major histology groups. Neuro Oncol 3(3):152–158

    PubMed  CAS  Google Scholar 

  2. Surawicz TS et al (1998) Brain tumor survival: results from the National Cancer Data Base. J Neurooncol 40(2):151–160

    Article  PubMed  CAS  Google Scholar 

  3. Zhang JG et al (2007) Antigenic profiling of glioma cells to generate allogeneic vaccines or dendritic cell-based therapeutics. Clin Cancer Res 13(2 Pt 1):566–575

    Article  PubMed  CAS  Google Scholar 

  4. Saikali S et al (2007) Expression of nine tumour antigens in a series of human glioblastoma multiforme: interest of EGFRvIII, IL-13Ralpha2, gp100 and TRP-2 for immunotherapy. J Neurooncol 81(2):139–148

    Article  PubMed  CAS  Google Scholar 

  5. Sampson JH et al (1996) Subcutaneous vaccination with irradiated, cytokine-producing tumor cells stimulates CD8+ cell-mediated immunity against tumors located in the “immunologically privileged” central nervous system. Proc Natl Acad Sci USA 93(19):10399–10404

    Article  PubMed  CAS  Google Scholar 

  6. Yu JS et al (2001) Vaccination of malignant glioma patients with peptide-pulsed dendritic cells elicits systemic cytotoxicity and intracranial T-cell infiltration. Cancer Res 61(3):842–847

    PubMed  CAS  Google Scholar 

  7. Mitchell DA, Fecci PE, Sampson JH (2008) Immunotherapy of malignant brain tumors. Immunol Rev 222:70–100

    Article  PubMed  CAS  Google Scholar 

  8. Liau LM et al (2005) Dendritic cell vaccination in glioblastoma patients induces systemic and intracranial T-cell responses modulated by the local central nervous system tumor microenvironment. Clin Cancer Res 11(15):5515–5525

    Article  PubMed  CAS  Google Scholar 

  9. Merchant RE et al (1988) Intralesional infusion of lymphokine-activated killer (LAK) cells and recombinant interleukin-2 (rIL-2) for the treatment of patients with malignant brain tumor. Neurosurgery 23(6):725–732

    Article  PubMed  CAS  Google Scholar 

  10. Lillehei KO et al (1991) Long-term follow-up of patients with recurrent malignant gliomas treated with adjuvant adoptive immunotherapy. Neurosurgery 28(1):16–23

    Article  PubMed  CAS  Google Scholar 

  11. Dillman RO et al (2004) Intracavitary placement of autologous lymphokine-activated killer (LAK) cells after resection of recurrent glioblastoma. J Immunother 27(5):398–404

    Article  PubMed  Google Scholar 

  12. Serraino D et al (2005) Infection with Epstein-Barr virus and cancer: an epidemiological review. J Biol Regul Homeost Agents 19(1–2):63–70

    PubMed  CAS  Google Scholar 

  13. Eiben GL et al (2003) Cervical cancer vaccines: recent advances in HPV research. Viral Immunol 16(2):111–121

    Article  PubMed  CAS  Google Scholar 

  14. Bosch FX et al (2005) Epidemiology of hepatocellular carcinoma. Clin Liver Dis 9(2):191–211, v

    Article  PubMed  Google Scholar 

  15. Vilchez RA, Butel JS (2003) Simian virus 40 and its association with human lymphomas. Curr Oncol Rep 5(5):372–379

    Article  PubMed  Google Scholar 

  16. Harkins L et al (2002) Specific localisation of human cytomegalovirus nucleic acids and proteins in human colorectal cancer. Lancet 360(9345):1557–1563

    Article  PubMed  CAS  Google Scholar 

  17. Cobbs CS et al (2002) Human cytomegalovirus infection and expression in human malignant glioma. Cancer Res 62(12):3347–3350

    PubMed  CAS  Google Scholar 

  18. Samanta M et al (2003) High prevalence of human cytomegalovirus in prostatic intraepithelial neoplasia and prostatic carcinoma. J Urol 170(3):998–1002

    Article  PubMed  Google Scholar 

  19. Mitchell DA et al (2008) Sensitive detection of human cytomegalovirus in tumors and peripheral blood of patients diagnosed with glioblastoma. Neuro Oncol 10(1):10–18

    Article  PubMed  Google Scholar 

  20. Scheurer ME et al (2008) Detection of human cytomegalovirus in different histological types of gliomas. Acta Neuropathol 116(1):79–86

    Article  PubMed  CAS  Google Scholar 

  21. Poltermann S et al (2006) Lack of association of herpesviruses with brain tumors. J Neurovirol 12(2):90–99

    Article  PubMed  CAS  Google Scholar 

  22. Lau SK et al (2005) Lack of association of cytomegalovirus with human brain tumors. Mod Pathol 18(6):838–843

    Article  PubMed  CAS  Google Scholar 

  23. Cinatl J et al (2008) Oncomodulatory signals by regulatory proteins encoded by human cytomegalovirus: a novel role for a viral infection in tumor progression. FEMS Microbiol Rev 28:59–77

    Article  Google Scholar 

  24. Hagemeier C et al (1994) Functional interaction between the HCMV IE2 transactivator and the retinoblastoma protein. EMBO J 13(12):2897–2903

    PubMed  CAS  Google Scholar 

  25. Fortunato EA et al (1997) Identification of domains within the human cytomegalovirus major immediate-early 86-kilodalton protein and the retinoblastoma protein required for physical and functional interaction with each other. J Virol 71(11):8176–8185

    PubMed  CAS  Google Scholar 

  26. Tsai HL et al (1996) Human cytomegalovirus immediate-early protein IE2 tethers a transcriptional repression domain to p53. J Biol Chem 271(7):3534–3540

    Article  PubMed  CAS  Google Scholar 

  27. Bao L et al (2008) Expansion of cytomegalovirus pp65 and IE-1 specific cytotoxic T lymphocytes for cytomegalovirus-specific immunotherapy following allogeneic stem cell transplantation. Biol Blood Marrow Transplant 14(10):1156–1162

    Article  PubMed  Google Scholar 

  28. Sun Q et al (2000) B lymphoblastoid cell lines as efficient APC to elicit CD8+ T cell responses against a cytomegalovirus antigen. J Immunol 165(7):4105–4111

    PubMed  CAS  Google Scholar 

  29. Yang I et al (2004) Modulation of major histocompatibility complex Class I molecules and major histocompatibility complex-bound immunogenic peptides induced by interferon-alpha and interferon-gamma treatment of human glioblastoma multiforme. J Neurosurg 100(2):310–319

    Article  PubMed  CAS  Google Scholar 

  30. Read SB et al (2003) Human alloreactive CTL interactions with gliomas and with those having upregulated HLA expression from exogenous IFN-gamma or IFN-gamma gene modification. J Interferon Cytokine Res 23(7):379–393

    Article  PubMed  CAS  Google Scholar 

  31. Serrano A et al (2001) Rexpression of HLA class I antigens and restoration of antigen-specific CTL response in melanoma cells following 5-aza-2′-deoxycytidine treatment. Int J Cancer 94(2):243–251

    Article  PubMed  CAS  Google Scholar 

  32. Ogura T et al (1986) Human cytomegalovirus persistent infection in a human central nervous system cell line: production of a variant virus with different growth characteristics. J Gen Virol 67(Pt 12):2605–2616

    Article  PubMed  Google Scholar 

  33. Luo MH, Fortunato EA (2007) Long-term infection and shedding of human cytomegalovirus in T98G glioblastoma cells. J Virol 81(19):10424–10436

    Article  PubMed  CAS  Google Scholar 

  34. Sabatier J et al (2005) Detection of human cytomegalovirus genome and gene products in central nervous system tumours. Br J Cancer 92(4):747–750

    Article  PubMed  CAS  Google Scholar 

  35. Staras SA et al (2006) Seroprevalence of cytomegalovirus infection in the United States, 1988–1994. Clin Infect Dis 43(9):1143–1151

    Article  PubMed  Google Scholar 

  36. Boeke CE et al (2008) CMV antibody prevalence and seroincidence in plateletpheresis donors. J Clin Apher 23(2):63–65

    Article  PubMed  Google Scholar 

  37. Thurner B et al (1999) Vaccination with mage-3A1 peptide-pulsed mature, monocyte-derived dendritic cells expands specific cytotoxic T cells and induces regression of some metastases in advanced stage IV melanoma. J Exp Med 190(11):1669–1678

    Article  PubMed  CAS  Google Scholar 

  38. Coulie PG et al (2002) Cytolytic T-cell responses of cancer patients vaccinated with a MAGE antigen. Immunol Rev 188:33–42

    Article  PubMed  CAS  Google Scholar 

  39. Ho WY et al (2006) In vitro methods for generating CD8+ T-cell clones for immunotherapy from the naive repertoire. J Immunol Methods 310(1–2):40–52

    Article  PubMed  CAS  Google Scholar 

  40. Borysiewicz LK et al (1988) Human cytomegalovirus-specific cytotoxic T cells. Relative frequency of stage-specific CTL recognizing the 72-kD immediate early protein and glycoprotein B expressed by recombinant vaccinia viruses. J Exp Med 168(3):919–931

    Article  PubMed  CAS  Google Scholar 

  41. Liu YN, Kari B, Gehrz RC (1988) Human immune responses to major human cytomegalovirus glycoprotein complexes. J Virol 62(3):1066–1070

    PubMed  CAS  Google Scholar 

  42. Ploegh HL (1998) Viral strategies of immune evasion. Science 280(5361):248–253

    Article  PubMed  CAS  Google Scholar 

  43. Oshiro S et al (2000) Response of MHC class-1 antigen on rat glioma cells to cytokines. Anticancer Res 20(1C):605–610

    PubMed  CAS  Google Scholar 

  44. Piguet V et al (1986) Heterogeneity of the induction of HLA-DR expression by human immune interferon on glioma cell lines and their clones. J Natl Cancer Inst 76(2):223–228

    PubMed  CAS  Google Scholar 

  45. Wen PY, Lampson MA, Lampson LA (1992) Effects of gamma-interferon on major histocompatibility complex antigen expression and lymphocytic infiltration in the 9L gliosarcoma brain tumor model: implications for strategies of immunotherapy. J Neuroimmunol 36(1):57–68

    Article  PubMed  CAS  Google Scholar 

  46. Mitchell D et al (2008) Efficacy of a phase II vaccine targeting cytomegalovirus antigens in newly-diagnosed GBM. J Clin Oncol 26:2042

    Google Scholar 

  47. Prins RM, Cloughesy TF, Liau LM (2008) Cytomegalovirus immunity after vaccination with autologous glioblastoma lysate. N Engl J Med 359(5):539–541

    Article  PubMed  CAS  Google Scholar 

  48. Bao L, Sun Q, Lucas KG (2007) Rapid generation of CMV pp65-specific T cells for immunotherapy. J Immunother (1997) 30(5):557–561

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pediatrics, Penn State Hershey Children’s Hospital, 500 University Drive, Room C7830, Hershey, PA, 17033, USA

    Kenneth G. Lucas, Lei Bao, Richard Bruggeman, Kimberly Dunham & Charles Specht

  2. Department of Pathology, Penn State Hershey Medical Center, Hershey, USA

    Kenneth G. Lucas, Lei Bao, Richard Bruggeman, Kimberly Dunham & Charles Specht

Authors
  1. Kenneth G. Lucas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lei Bao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Richard Bruggeman
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kimberly Dunham
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Charles Specht
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kenneth G. Lucas.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lucas, K.G., Bao, L., Bruggeman, R. et al. The detection of CMV pp65 and IE1 in glioblastoma multiforme. J Neurooncol 103, 231–238 (2011). https://doi.org/10.1007/s11060-010-0383-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11060-010-0383-6

Keywords

Search

Navigation