Abstract
The role of the human cytomegalovirus (HCMV) in gliomagenesis is largely debated. Contradictory data exist regarding the sensitivity and specificity of HCMV detection techniques, including immunohistochemistry (IHC), in situ hybridization (ISH), and RNA and DNA sequencing. The aim of this study is to detect HCMV in glioblastoma (GBM) tumor samples using IHC, ISH, and real-time PCR (qPCR), as well as to correlate the findings with serological status and HCMV DNA load in blood. Forty-seven patients with histopathological diagnosis of GBM and HCMV serological status were retrospectively reviewed. HCMV DNA quantification in whole blood was performed in 31 patients. The detection of HCMV in tumor samples was performed using IHC in 42 cases, ISH in 10 cases, and qPCR in 29 cases. All but two patients were taking high steroid doses at the time of biological testing. HCMV seroprevalence was 68%. Active infection with HCMV DNA detected in blood was diagnosed in 6 out of 21 (28%) seropositive patients. HCMV was not detected in GBM samples using IHC or ISH, while qPCR was positive in one case (also positive for blood HCMV DNA). These data do not support a crucial role of HCMV in GBM tumorigenesis. HCMV might be reactivated in GBM patients, due to steroid treatment.
Similar content being viewed by others
References
Adle-Biassette H, Grassi J, Verney C, Walker F, Choudat L, Hénin D (2007) The importance of controls in immunohistochemistry: how to validate an antibody, from research to diagnosis. Ann Pathol 27:16–26
Baumgarten P, Michaelis M, Rothweiler F, Starzetz T, Rabenau HF, Berger A, Jennewein L, Braczynski AK, Franz K, Seifert V, Steinbach JP, Allwinn R, Mittelbronn M, Cinatl J (2014) Human cytomegalovirus infection in tumor cells of the nervous system is not detectable with standardized pathologico-virological diagnostics. Neuro Oncol 16:1469–1477. https://doi.org/10.1093/neuonc/nou167
Cannon MJ, Schmid DS, Hyde TB (2010) Review of cytomegalovirus seroprevalence and demographic characteristics associated with infection. Rev Med Virol 20:202–213. https://doi.org/10.1002/rmv.655
Chen H-P, Jiang J-K, Chan C-H et al (2015) Genetic polymorphisms of the human cytomegalovirus UL144 gene in colorectal cancer and its association with clinical outcome. J Gen Virol 96:3613–3623. https://doi.org/10.1099/jgv.0.000308
Cobbs C (2014) Response to “human cytomegalovirus infection in tumor cells of the nervous system is not detectable with standardized pathologico-virological diagnostics.”. Neuro Oncol 16:1435–1436. https://doi.org/10.1093/neuonc/nou295
Cobbs CS, Harkins L, Samanta M, Gillespie GY, Bharara S, King PH, Nabors LB, Cobbs CG, Britt WJ (2002) Human cytomegalovirus infection and expression in human malignant glioma. Cancer Res 62:3347–3350
Cosset É, Petty TJ, Dutoit V, Cordey S, Padioleau I, Otten-Hernandez P, Farinelli L, Kaiser L, Bruyère-Cerdan P, Tirefort D, Amar el-Dusouqui S, Nayernia Z, Krause KH, Zdobnov EM, Dietrich PY, Rigal E, Preynat-Seauve O (2014) Comprehensive metagenomic analysis of glioblastoma reveals absence of known virus despite antiviral-like type I interferon gene response. Int J Cancer 135:1381–1389. https://doi.org/10.1002/ijc.28670
Crough T, Khanna R (2009) Immunobiology of human cytomegalovirus: from bench to bedside. Clin Microbiol Rev 22:76–98. https://doi.org/10.1128/CMR.00034-08
Dziurzynski K, Chang SM, Heimberger AB, Kalejta RF, McGregor Dallas SR, Smit M, Soroceanu L, Cobbs CS, the HCMV and Gliomas Symposium (2012) Consensus on the role of human cytomegalovirus in glioblastoma. Neuro Oncol 14:246–255. https://doi.org/10.1093/neuonc/nor227
Garcia-Martinez A, Alenda C, Irles E, Ochoa E, Quintanar T, Rodriguez-Lescure A, Soto JL, Barbera VM (2017) Lack of cytomegalovirus detection in human glioma. Virol J 14:216. https://doi.org/10.1186/s12985-017-0885-3
Goerig NL, Frey B, Überla K, Gaipl U, Fietkau R (2017) A clinician’s plea to test glioma patients for CMV. Neuro Oncol 19:1282–1283. https://doi.org/10.1093/neuonc/nox080
Hochhalter CB, Carr C, O’Neill BE, Ware ML, Strong MJ (2017) The association between human cytomegalovirus and glioblastomas: a review. Neuroimmunol Neuroinflammation 4:96. https://doi.org/10.20517/2347-8659.2017.10
Holdhoff M, Guner G, Rodriguez FJ, Hicks JL, Zheng Q, Forman MS, Ye X, Grossman SA, Meeker AK, Heaphy CM, Eberhart CG, de Marzo AM, Arav-Boger R (2017) Absence of cytomegalovirus in glioblastoma and other high-grade gliomas by real-time PCR, immunohistochemistry, and in situ hybridization. Clin Cancer Res 23:3150–3157. https://doi.org/10.1158/1078-0432.CCR-16-1490
Johnson DR, O’Neill BP (2012) Glioblastoma survival in the United States before and during the temozolomide era. J Neuro-Oncol 107:359–364. https://doi.org/10.1007/s11060-011-0749-4
Johnson TS, Abrams ZB, Mo X, Zhang Y, Huang K (2017) Lack of human cytomegalovirus expression in single cells from glioblastoma tumors and cell lines. J Neuro-Oncol 23:671–678. https://doi.org/10.1007/s13365-017-0543-y
Khoury JD, Tannir NM, Williams MD, Chen Y, Yao H, Zhang J, Thompson EJ, the TCGA Network, Meric-Bernstam F, Medeiros LJ, Weinstein JN, Su X (2013) Landscape of DNA virus associations across human malignant cancers: analysis of 3,775 cases using RNA-Seq. J Virol 87:8916–8926. https://doi.org/10.1128/JVI.00340-13
Lau SK, Chen Y-Y, Chen W-G, Diamond DJ, Mamelak AN, Zaia JA, Weiss LM (2005) Lack of association of cytomegalovirus with human brain tumors. Mod Pathol 18:838–843. https://doi.org/10.1038/modpathol.3800352
Lin C-TM, Leibovitch EC, Almira-Suarez MI, Jacobson S (2016) Human herpesvirus multiplex ddPCR detection in brain tissue from low- and high-grade astrocytoma cases and controls. Infect Agent Cancer 11:32. https://doi.org/10.1186/s13027-016-0081-x
Ljungman P (2014) The role of cytomegalovirus serostatus on outcome of hematopoietic stem cell transplantation. Curr Opin Hematol 21:466–469. https://doi.org/10.1097/MOH.0000000000000085
Louis DN, Perry A, Reifenberger G, von Deimling A, Figarella-Branger D, Cavenee WK, Ohgaki H, Wiestler OD, Kleihues P, Ellison DW (2016) The 2016 World Health Organization classification of tumors of the central nervous system: a summary. Acta Neuropathol (Berl) 131:803–820. https://doi.org/10.1007/s00401-016-1545-1
Löwenberg M, Stahn C, Hommes DW, Buttgereit F (2008) Novel insights into mechanisms of glucocorticoid action and the development of new glucocorticoid receptor ligands. Steroids 73:1025–1029. https://doi.org/10.1016/j.steroids.2007.12.002
Michaelis M, Mittelbronn M, Cinatl J (2015) Towards an unbiased, collaborative effort to reach evidence about the presence of human cytomegalovirus in glioblastoma (and other tumors). Neuro Oncol 17:1039. https://doi.org/10.1093/neuonc/nov048
Richardson AK, Currie MJ, Robinson BA, Morrin H, Phung Y, Pearson JF, Anderson TP, Potter JD, Walker LC (2015) Cytomegalovirus and Epstein-Barr virus in breast cancer. PLoS One 10:e0118989. https://doi.org/10.1371/journal.pone.0118989
Saper CB (2005) An open letter to our readers on the use of antibodies. J Comp Neurol 493:477–478. https://doi.org/10.1002/cne.20839
Saper CB, Sawchenko PE (2003) Magic peptides, magic antibodies: guidelines for appropriate controls for immunohistochemistry. J Comp Neurol 465:161–163. https://doi.org/10.1002/cne.10858
Solomon IH, Ramkissoon SH, Milner DA, Folkerth RD (2014) Cytomegalovirus and glioblastoma: a review of evidence for their association and indications for testing and treatment. J Neuropathol Exp Neurol 73:994–998. https://doi.org/10.1097/NEN.0000000000000125
Staras SAS, Dollard SC, Radford KW, Flanders WD, Pass RF, Cannon MJ (2006) Seroprevalence of cytomegalovirus infection in the United States, 1988-1994. Clin Infect Dis 43:1143–1151. https://doi.org/10.1086/508173
Strong MJ, Blanchard E, Lin Z, Morris CA, Baddoo M, Taylor CM, Ware ML, Flemington EK (2016) A comprehensive next generation sequencing-based virome assessment in brain tissue suggests no major virus - tumor association. Acta Neuropathol Commun 4:71. https://doi.org/10.1186/s40478-016-0338-z
Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, Curschmann J, Janzer RC, Ludwin SK, Gorlia T, Allgeier A, Lacombe D, Cairncross JG, Eisenhauer E, Mirimanoff RO, European Organisation for Research and Treatment of Cancer Brain Tumor and Radiotherapy Groups, National Cancer Institute of Canada Clinical Trials Group (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352:987–996. https://doi.org/10.1056/NEJMoa043330
Taha MS, Abdalhamid BA, El-Badawy SA et al (2016) Expression of cytomegalovirus in glioblastoma multiforme: myth or reality? Br J Neurosurg 30:307–312. https://doi.org/10.3109/02688697.2015.1119241
Tang K-W, Alaei-Mahabadi B, Samuelsson T, Lindh M, Larsson E (2013) The landscape of viral expression and host gene fusion and adaptation in human cancer. Nat Commun 4:2513. https://doi.org/10.1038/ncomms3513
Tang K-W, Hellstrand K, Larsson E (2015) Absence of cytomegalovirus in high-coverage DNA sequencing of human glioblastoma multiforme. Int J Cancer 136:977–981. https://doi.org/10.1002/ijc.29042
Teissier N, Delezoide A-L, Mas A-E, Khung-Savatovsky S, Bessières B, Nardelli J, Vauloup-Fellous C, Picone O, Houhou N, Oury JF, van den Abbeele T, Gressens P, Adle-Biassette H (2011) Inner ear lesions in congenital cytomegalovirus infection of human fetuses. Acta Neuropathol (Berl) 122:763–774. https://doi.org/10.1007/s00401-011-0895-y
Teissier N, Fallet-Bianco C, Delezoide A-L, Laquerrière A, Marcorelles P, Khung-Savatovsky S, Nardelli J, Cipriani S, Csaba Z, Picone O, Golden JA, van den Abbeele T, Gressens P, Adle-Biassette H (2014) Cytomegalovirus-induced brain malformations in fetuses. J Neuropathol Exp Neurol 73:143–158. https://doi.org/10.1097/NEN.0000000000000038
Ward JM (2004) Controls for immunohistochemistry: is “brown” good enough? Toxicol Pathol 32:273–274. https://doi.org/10.1080/01926230490457585
Wen PY, Kesari S (2008) Malignant gliomas in adults. N Engl J Med 359:492–507. https://doi.org/10.1056/NEJMra0708126
Yamashita Y, Ito Y, Isomura H, Takemura N, Okamoto A, Motomura K, Tsujiuchi T, Natsume A, Wakabayashi T, Toyokuni S, Tsurumi T (2014) Lack of presence of the human cytomegalovirus in human glioblastoma. Mod Pathol 27:922–929. https://doi.org/10.1038/modpathol.2013.219
Acknowledgments
Authors thank Diana Folia and Dr. Darren Russell for their careful English editing of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Written consent for use of molecular analysis for research purpose was obtained and recorded in each patient’s medical file. This study was approved by the local Ethics Committee of the Neuroscience division of Lariboisière Hospital.
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Emmanuel Mandonnet and Jean-Michel Molina are co-last authors.
Rights and permissions
About this article
Cite this article
Loit, MP., Adle-Biassette, H., Bouazza, S. et al. Multimodal techniques failed to detect cytomegalovirus in human glioblastoma samples. J. Neurovirol. 25, 50–56 (2019). https://doi.org/10.1007/s13365-018-0683-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13365-018-0683-8