Susceptibility of rat immortalized neuronal cell line Rn33B expressing equine major histocompatibility class 1 to equine herpesvirus-1 infection is differentiation dependent.
Abstract: Equine herpesvirus-1 (EHV-1), which causes encephalomyelitis in horses, shows endotheliotropism in the central nervous system of horses, and generally does not infect neurons. However, little is known about the mechanism underlying the resistance of neuron to EHV-1, due to the lack of convenient cell culture systems. In this study, we examined EHV-1 infection in immortalized Rn33B rat neuronal cells, which differentiate into neurons when cultured under nonpermissive conditions. Because murine cell lines are resistant to EHV-1 infections due to the lack of functional entry receptors for EHV-1, we used an Rn33B-derived cell line that stably expresses the equine MHC class 1 molecule, which acts as EHV-1 entry receptor (Rn33B-A68B2M cells). EHV-1 infected undifferentiated Rn33B-A68B2M cells more efficiently than differentiated cells, resulting in the production of progeny virus in the former but not in the latter. By contrast, both differentiated and undifferentiated cells infected with herpes simplex virus-1 produced infectious viral progeny. While EHV-1 infection induced stronger expression of IFN alpha gene in differentiated cells than in undifferentiated cells, downstream IFN responses, including phosphorylation of STAT1 (signal transducer and activator of transcription 1) and expression of IFN-stimulated genes, were not activated regardless of whether cells were differentiated or not. These results suggest that neuronal differentiation of RN33B-A68B2M cells reduced their susceptibility to EHV-1, which is not due to different IFN responses. This culture system may be useful as an in vitro model for studying neuron-specific resistance to EHV-1, by investigating viral and host factors responsible for the difference in susceptibility between differentiated and undifferentiated cells.
© 2019 The Societies and John Wiley & Sons Australia, Ltd.
Publication Date: 2020-01-21 PubMed ID: 31758567DOI: 10.1111/1348-0421.12761Google Scholar: Lookup
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- Journal Article
Summary
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The research investigated the effects of equine herpesvirus-1 (EHV-1) on differentiated and undifferentiated immortalized rat neuronal cells (Rn33B), finding that the differentiation of these cells reduces their susceptibility to the virus.
Context and Research Objective
- The research aimed to understand why equine herpesvirus-1 (EHV-1), a virus causing encephalomyelitis in horses, typically infects endothelial cells in the central nervous system (CNS) of horses but not neurons. Previous research was limited by a lack of convenient cell culture systems.
- The Rn33B rat neuronal cell line was used in this study as it differentiates into neurons under nonpermissive conditions, providing a useful model for studying neuron-specific resistance to EHV-1.
- An additional objective was to investigate the role of the equine major histocompatibility class 1 (MHC class 1) molecule which acts as an entry receptor for EHV-1 infection.
Methodology
- The research used an Rn33B-derived cell line that stably expresses the equine MHC class 1 molecule, known as Rn33B-A68B2M cells.
- These cells were exposed to EHV-1 infection, and the impacts on differentiated and undifferentiated cells were compared. Herpes simplex virus-1 (HSV-1) was also used as a control.
Key Findings
- The study found that undifferentiated Rn33B-A68B2M cells were infected by EHV-1 more efficiently than differentiated cells. The undifferentiated cells produced offsprings of the virus, while the differentiated cells did not.
- The comparison with HSV-1 infection showed that either differentiated or undifferentiated cells produced infectious viral offspring, indicating that the effect seen with EHV-1 was particular to this virus.
- The results also showed that EHV-1 infection induced a stronger expression of the IFN alpha gene in differentiated cells than in undifferentiated cells. However, downstream IFN responses, such as phosphorylation of STAT1 and expression of IFN-stimulated genes, were not activated in either type of cell.
Implications and Further Research
- The findings indicate that the differentiation of RN33B-A68B2M cells reduces their susceptibility to EHV-1, but this is not due to different IFN responses.
- The cell culture system used in this study could provide a valuable in-vitro model for future research into the neuron-specific resistance to EHV-1. This could involve investigating viral and host factors responsible for the difference in susceptibility between differentiated and undifferentiated cells.
Cite This Article
APA
Minato E, Kobayashi A, Aoshima K, Fukushi H, Kimura T.
(2020).
Susceptibility of rat immortalized neuronal cell line Rn33B expressing equine major histocompatibility class 1 to equine herpesvirus-1 infection is differentiation dependent.
Microbiol Immunol, 64(2), 123-132.
https://doi.org/10.1111/1348-0421.12761 Publication
Researcher Affiliations
- Laboratory of Comparative Pathology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido, Japan.
- Laboratory of Comparative Pathology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido, Japan.
- Laboratory of Comparative Pathology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido, Japan.
- Laboratory of Veterinary Microbiology, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan.
- Laboratory of Comparative Pathology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido, Japan.
MeSH Terms
- Animals
- Cell Differentiation
- Cell Line
- Encephalomyelitis / veterinary
- Encephalomyelitis / virology
- Herpesviridae Infections
- Herpesvirus 1, Equid / pathogenicity
- Histocompatibility Antigens Class I / metabolism
- Horse Diseases / virology
- Horses
- Immediate-Early Proteins / metabolism
- Interferons / metabolism
- Mice
- Neurons / metabolism
- Neurons / virology
- Rats
- Virus Internalization
Grant Funding
- Grant-in-Aid for Scientific Research (B)/16H05022 / Ministry of Education, Culture, Sports, Science and Technology
References
This article includes 23 references
- Ma G, Azab W, Osterrieder N. Equine herpesviruses type 1 (EHV-1) and 4 (EHV-4)-Masters of co-evolution and a constant threat to equids and beyond.. Vet Microbiol 2013;167:123-34.
- Danaher RJ, Jacob RJ, Miller CS. Establishment of a quiescent herpes simplex virus type 1 infection in neurally-differentiated PC12 cells.. J Neurovirol 1999;5:258-67.
- Danaher RJ, Jacob RJ, Steiner MR, Allen WR, Hill JM, Miller CS. Histone deacetylase inhibitors induce reactivation of herpes simplex virus type 1 in a latency-associated transcript (LAT)-independent manner in neuronal cells.. J Neurovirol 2005;11:306-17.
- Jiang X, Chentoufi AA, Hsiang C. The herpes simplex virus type 1 latency-associated transcript can protect neuron-derived C1300 and Neuro2A cells from granzyme B-induced apoptosis and CD8 T-cell killing.. J Virol 2011;85:2325-32.
- Sasaki M, Hasebe R, Makino Y. Equine major histocompatibility complex class I molecules act as entry receptors that bind to equine herpesvirus-1 glycoprotein D.. Genes Cells 2011;16:343-57.
- Whittemore SR, White LA. Target regulation of neuronal differentiation in a temperature-sensitive cell line derived from medullary raphe.. Brain Res 1993;615:27-40.
- Sasaki M, Kim E, Igarashi M. Single amino acid residue in the A2 domain of major histocompatibility complex class I is involved in the efficiency of equine herpesvirus-1 entry.. J Biol Chem 2011;286:39370-8.
- Wysocka J, Herr W. The herpes simplex virus VP16-induced complex: the makings of a regulatory switch.. Trends Biochem Sci 2003;28:294-304.
- Nugent J, Birch-Machin I, Smith KC. Analysis of equid herpesvirus 1 strain variation reveals a point mutation of the DNA polymerase strongly associated with neuropathogenic versus nonneuropathogenic disease outbreaks.. J Virol 2006;80:4047-60.
- Ibrahim el SM, Pagmajav O, Yamaguchi T, Matsumura T, Fukushi H. Growth and virulence alterations of equine herpesvirus 1 by insertion of a green fluorescent protein gene in the intergenic region between ORFs 62 and 63.. Microbiol Immunol 2004;48:831-42.
- Minato E, Aoshima K, Kobayashi A, Ohnishi N, Sasaki N, Kimura T. Exogenous expression of equine MHC class I molecules in mice increases susceptibility to equine herpesvirus 1 pulmonary infection.. Vet Pathol 2019;56:703-10.
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method.. Methods 2001;25:402-8.
- Hasebe R, Kimura T, Nakamura K. Differential susceptibility of equine and mouse brain microvascular endothelial cells to equine herpesvirus 1 infection.. Arch Virol 2006;151:775-86.
- Kimura T, Okumura M, Kim E, Sasaki M, Orba Y, Sawa H. Characterization of Japanese encephalitis virus infection in an immortalized mesencephalic cell line, CSM14.1.. Microbiol Immunol 2013;57:723-31.
- Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis.. Nat Methods 2012;9:671-5.
- Hudson SJ, Dix RD, Streilein JW. Induction of encephalitis in SJL mice by intranasal infection with herpes simplex virus type 1: a possible model of herpes simplex encephalitis in humans.. J Infect Dis 1991;163:720-7.
- Drummond CW, Eglin RP, Esiri MM. Herpes simplex virus encephalitis in a mouse model: PCR evidence for CNS latency following acute infection.. J Neurol Sci 1994;127:159-63.
- Kristie TM, Vogel JL, Sears AE. Nuclear localization of the C1 factor (host cell factor) in sensory neurons correlates with reactivation of herpes simplex virus from latency.. Proc Natl Acad Sci U S A 1999;96:1229-33.
- Patel JR, Edington N. The pathogenicity in mice of respiratory, abortion and paresis isolates of equine herpesvirus-1.. Vet Microbiol 1983;8:301-5.
- Nowotny N, Burtscher H, Bürki F. Neuropathogenicity for suckling mice of equine herpesvirus 1 from the Lipizzan outbreak 1983 and of selected other EHV 1 strains.. J Vet Med Ser B 1987;34:441-8.
- Bartels T, Steinbach F, Hahn G, Ludwig H, Borchers K. In situ study on the pathogenesis and immune reaction of equine herpesvirus type 1 (EHV-1) infections in mice.. Immunology 1998;93:329-34.
- Walker C, Love DN, Whalley JM. Comparison of the pathogenesis of acute equine herpesvirus 1 (EHV-1) infection in the horse and the mouse model: a review.. Vet Microbiol 1999;68:3-13.
- Gosztonyi G, Borchers K, Ludwig H. Pathogenesis of equine herpesvirus-1 infection in the mouse model.. APMIS 2009;117:10-21.
Citations
This article has been cited 1 times.- Maeda M, Abe M, Aoshima K, Kobayashi A, Fukushi H, Kimura T. Identification of the Promoter Antisense Transcript Enhancing the Transcription of the Equine Herpesvirus-1 Immediate-Early Gene. Viruses 2024 Jul 25;16(8).
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