Abstract
Cytomegalovirus (CMV) is the most common cause of congenital infection in developed countries and a major cause of neurological disability in children. Although CMV can affect multiple organs, the most important sequelae of intrauterine infection are related to lesions of the central nervous system. However, little is known about the pathogenesis and the cellular events responsible for neuronal damage in infants with congenital infection. Some studies have demonstrated that neural precursor cells (NPCs) show the greatest susceptibility to CMV infection in the developing brain. We sought to establish an in vitro model of CMV infection of the developing brain in order to analyze the cellular events associated with invasion by this virus. To this end, we employed two cell lines as a permanent source of NPC, avoiding the continuous use of human fetal tissue, the human SK-N-MC neuroblastoma cell line, and an immortalized cell line of human fetal neural origin, hNS-1. We also investigated the effect of the differentiation stage in relation to the susceptibility of these cell lines by comparing the neuroblastoma cell line with the multipotent cell line hNS-1. We found that the effects of the virus were more severe in the neuroblastoma cell line. Additionally, we induced hNS-1 to differentiate and evaluated the effect of CMV in these differentiated cells. Like SK-N-MC cells, hNS-1-differentiated cells were also susceptible to infection. Viability of differentiated hNS-1 cells decreased after CMV infection in contrast to undifferentiated cells. In addition, differentiated hNS-1 cells showed an extensive cytopathic effect whereas the effect was scarce in undifferentiated cells. We describe some of the effects of CMV in neural stem cells, and our observations suggest that the degree of differentiation is important in the acquisition of susceptibility.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Agol VI (2012) Cytopathic effects: virus-modulated manifestations of innate immunity? Trends Microbiol 20(12):570–576
Belec L, Mhiri C, Belghiti D, Geny C, Boudes P, Gray F (1990) Cytomegalovirus (CMV) and human immunodeficiency virus (HIV) co-infection, of multinucleated giant cells in acquired immunodeficiency syndrome (AIDS) encephalopathy. Arch Anat Cytol Pathol 38:189–197
Cheeran MC, Hu S, Ni HT, Sheng W, Palmquist JM, Peterson PK, Lokensgard JR (2005) Neural precursor cell susceptibility to human cytomegalovirus diverges along glial or neuronal differentiation pathways. J Neurosci Res 82:839–850
Cheeran MC, Lokensgard JR, Schleiss MR (2009) Neuropathogenesis of congenital cytomegalovirus infection: disease mechanisms and prospects for intervention. Clin Microbiol Rev 22:99–126, Table of Contents
DeMarchi JM (1983) Nature of the block in the expression of some early virus genes in cells abortively infected with human cytomegalovirus. Virology 129:287–297
Griffiths PD, Grundy JE (1987) Molecular biology and immunology of cytomegalovirus. Biochem J 241:313–324
Higgins S, Wong SH, Richner M, Rowe CL, Newgreen DF, Werther GA, Russo VC (2009) Fibroblast growth factor 2 reactivates G1 checkpoint in SK-N-MC cells via regulation of p21, inhibitor of differentiation genes (Id1-3), and epithelium-mesenchyme transition-like events. Endocrinology 150:4044–4055
Ho M (2008) The history of cytomegalovirus and its diseases. Med Microbiol Immunol 197:65–73
Ho WS, van den Pol AN (2007) Bystander attenuation of neuronal and astrocyte intercellular communication by murine cytomegalovirus infection of glia. J Virol 81:7286–7292
Jafferji I, Bain M, King C, Sinclair JH (2009) Inhibition of epidermal growth factor receptor (EGFR) expression by human cytomegalovirus correlates with an increase in the expression and binding of Wilms’ Tumour 1 protein to the EGFR promoter. J Gen Virol 90:1569–1574
Kawasaki H, Tsutsui Y (2003) Brain slice culture for analysis of developmental brain disorders with special reference to congenital cytomegalovirus infection. Congenit Anom (Kyoto) 43:105–113
Kawasaki H, Kosugi I, Arai Y, Iwashita T, Tsutsui Y (2011) Mouse embryonic stem cells inhibit murine cytomegalovirus infection through a multi-step process. PLoS One 6:e17492
Kinzler ER, Compton T (2005) Characterization of human cytomegalovirus glycoprotein-induced cell-cell fusion. J Virol 79:7827–7837
Köhler TH (1995) Quantitative RT-PCR. In: Quantitation of mRNA by polymerase chain reaction (Springer-Verlag, ed), pp 71-80 Berlin Heidelberg
Luo MH, Schwartz PH, Fortunato EA (2008) Neonatal neural progenitor cells and their neuronal and glial cell derivatives are fully permissive for human cytomegalovirus infection. J Virol 82:9994–10007
Luo MH, Hannemann H, Kulkarni AS, Schwartz PH, O’Dowd JM, Fortunato EA (2010) Human cytomegalovirus infection causes premature and abnormal differentiation of human neural progenitor cells. J Virol 84:3528–3541
Martinez-Serrano A, Rubio FJ, Navarro B, Bueno C, Villa A (2001) Human neural stem and progenitor cells: in vitro and in vivo properties, and potential for gene therapy and cell replacement in the CNS. Curr Gene Ther 1(3):279–299
Matsukage S, Kosugi I, Kawasaski H, Miura K, Kitani H, Tsutsui Y (2006) Mouse embryonic stem cells are not susceptible to cytomegalovirus but acquire susceptibility during differentiation. Birth Defects Res A Clin Mol Teratol 76:115–125
Monsivais-Urenda A, Noyola-Cherpitel D, Hernandez-Salinas A, Garcia-Sepulveda C, Romo N, Baranda L, Lopez-Botet M, Gonzalez-Amaro R (2010) Influence of human cytomegalovirus infection on the NK cell receptor repertoire in children. Eur J Immunol 40:1418–1427
Navarro-Galve B, Villa A, Bueno C, Thompson L, Johansen J, Martinez-Serrano A (2005) Gene marking of human neural stem/precursor cells using green fluorescent proteins. J Gene Med 7(1):18–29
Noyola DE, Jimenez-Capdeville ME, Demmler-Harrison GJ (2010) Central nervous system disorders in infants with congenital cytomegalovirus infection. Neurol Res 32:278–284
Odeberg J, Wolmer N, Falci S, Westgren M, Seiger A, Soderberg-Naucler C (2006) Human cytomegalovirus inhibits neuronal differentiation and induces apoptosis in human neural precursor cells. J Virol 80:8929–8939
Odeberg J, Wolmer N, Falci S, Westgren M, Sundtrom E, Seiger A, Soderberg-Naucler C (2007) Late human cytomegalovirus (HCMV) proteins inhibit differentiation of human neural precursor cells into astrocytes. J Neurosci Res 85:583–593
Ornoy A (2007) Fetal effects of primary and non-primary cytomegalovirus infection in pregnancy: are we close to prevention? Isr Med Assoc J 9:398–401
Rubio FJ, Bueno C, Villa A, Navarro B, Martinez-Serrano A (2000) Genetically perpetuated human neural stem cells engraft and differentiate into the adult mammalian brain. Mol Cell Neurosci 16:1–13
Shinmura Y, Kosugi I, Aiba-Masago S, Baba S, Yong LR, Tsutsui Y (1997) Disordered migration and loss of virus-infected neuronal cells in developing mouse brains infected with murine cytomegalovirus. Acta Neuropathol 93:551–557
Tsutsui Y (2009) Effects of cytomegalovirus infection on embryogenesis and brain development. Congenit Anom (Kyoto) 49:47–55
Tsutsui Y, Kawasaki H, Kosugi I (2002) Reactivation of latent cytomegalovirus infection in mouse brain cells detected after transfer to brain slice cultures. J Virol 76:7247–7254
Tsutsui Y, Kosugi I, Kawasaki H, Arai Y, Han GP, Li L, Kaneta M (2008) Roles of neural stem progenitor cells in cytomegalovirus infection of the brain in mouse models. Pathol Int 58:257–267
van Den Pol AN, Mocarski E, Saederup N, Vieira J, Meier TJ (1999) Cytomegalovirus cell tropism, replication, and gene transfer in brain. J Neurosci 19:10948–10965
Villa A, Snyder EY, Vescovi A, Martinez-Serrano A (2000) Establishment and properties of a growth factor-dependent, perpetual neural stem cell line from the human CNS. Exp Neurol 161:67–84
Villa A, Rubio FJ, Navarro B, Bueno C, Martinez-Serrano A (2001) Human neural stem cells in vitro. A focus on their isolation and perpetuation. Biomed Pharmacother 55:91–95
Villa A, Navarro B, Martinez-Serrano A (2002) Genetic perpetuation of in vitro expanded human neural stem cells: cellular properties and therapeutic potential. Brain Res Bull 57:789–794
Villa A, Navarro-Galve B, Bueno C, Franco S, Blasco MA, Martinez-Serrano A (2004) Long-term molecular and cellular stability of human neural stem cell lines. Exp Cell Res 294:559–570
Wentworth BB, French L (1970) Plaque assay of cytomegalovirus strains of human origin. Proc Soc Exp Biol Med 135:253–258
Woolf NK, Jaquish DV, Koehrn FJ (2007) Transplacental murine cytomegalovirus infection in the brain of SCID mice. Virol J 4:26
Wynn KK, Khanna R (2006) Models of CMV infection. Drug Discov Today 3:91–96
Acknowledgments
This work was supported by grant from CONACYT (CB16782 and #120452), PROMEP (103.5/10/7697), and FAI-UASLP (C12-FAI-03-62.62). Hilda Gonzalez and Claudia Salazar were recipients of scholarships from CONACYT (CVU 265757 and 265189).
Conflict of interests
The authors declare that there is no conflict of interests regarding the publication of this paper.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
González-Sánchez, H.M., Monsiváis-Urenda, A., Salazar-Aldrete, C.A. et al. Effects of cytomegalovirus infection in human neural precursor cells depend on their differentiation state. J. Neurovirol. 21, 346–357 (2015). https://doi.org/10.1007/s13365-015-0315-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13365-015-0315-5