Regional and Temporal Patterns of Long-Term Pseudorabies Virus Detection and Neuropathology in the Murine CNS

Viktoria Korff; Issam El-Debs; Barbara G. Klupp; Conrad M. Freuling; Jens P. Teifke; Thomas C. Mettenleiter; Julia Sehl-Ewert

doi:10.3390/pathogens15040395

journal article Open Access Apr 07, 2026

Regional and Temporal Patterns of Long-Term Pseudorabies Virus Detection and Neuropathology in the Murine CNS

Viktoria Korff

Issam El-Debs

Barbara G. Klupp Conrad M. Freuling

Jens P. Teifke

Thomas C. Mettenleiter

Julia Sehl-Ewert

Pathogens Vol. 15 No. 4 pp. 395 · MDPI AG

View at Publisher Save 10.3390/pathogens15040395

Abstract

Alphaherpesviruses, including Herpes Simplex Virus 1 (HSV-1) and Pseudorabies Virus (PrV), establish lifelong latency in the nervous system and can cause recurrent disease. While latency has classically been attributed to peripheral sensory ganglia, accumulating evidence indicates that the central nervous system (CNS) may also serve as a site of long-term viral persistence and reactivation. Here, we investigated the CNS as a viral reservoir using the attenuated mutant PrV-∆UL21/US3∆kin, which preferentially targets mesiotemporal brain regions. Following intranasal inoculation, mice were analyzed at 11–14, 21, 28, 42, 105, and 190 days post-infection (dpi). To assess the reactivation potential, a subset of animals received cyclophosphamide/dexamethasone at 170 dpi. Viral transcripts were detected by RNAscope™ in situ hybridization and RT-qPCR targeting the lytic gene UL19 encoding the major capsid protein and the latency-associated transcript (LAT). Histopathology included hematoxylin and eosin staining and immunohistochemistry for CD3, Iba1, GFAP, cleaved caspase-3 and viral glycoprotein gB. UL19 RNA signals displayed marked regional and temporal heterogeneity, with prominent detection in mesiotemporal structures. In contrast, LAT RNA levels remained low overall, with a transient peak during the acute phase. RT-qPCR confirmed high UL19 and LAT transcript levels during early infection, while LAT transcription returned to baseline levels thereafter. Histopathology showed a transition from acute necrotizing meningoencephalitis to prolonged low-grade inflammation with glial activation and focal apoptosis. Notably, UL19 RNA signals strongly correlated with T-cell infiltration, particularly at 42 dpi. Together, these findings define regional and temporal patterns of long-term PrV transcriptional activity and associated neuropathology in the murine CNS.

Topics

No keywords indexed for this article. Browse by subject →

References

63

[1]

Koyuncu "Latent versus productive infection: The alpha herpesvirus switch" Future Virol. (2018) 10.2217/fvl-2018-0023

[2]

Spear "Three classes of cell surface receptors for alphaherpesvirus entry" Virology (2000) 10.1006/viro.2000.0529

[3]

Mettenleiter "Pathogenesis of neurotropic herpesviruses: Role of viral glycoproteins in neuroinvasion and transneuronal spread" Virus Res. (2003) 10.1016/s0168-1702(02)00352-0

[4]

Smith "Herpesvirus transport to the nervous system and back again" Annu. Rev. Microbiol. (2012) 10.1146/annurev-micro-092611-150051

[5]

Hill "Gene Expression Analyzed by Microarrays in HSV-1 Latent Mouse Trigeminal Ganglion Following Heat Stress" Virus Genes (2001) 10.1023/a:1012517221937

[6]

Hill "Acute and Recurrent Infection with Herpes Simplex Virus in the Mouse: A Model for Studying Latency and Recurrent Disease" J. Gen. Virol. (1975) 10.1099/0022-1317-28-3-341

[7]

Duarte, L.F., Farías, M.A., Álvarez, D.M., Bueno, S.M., Riedel, C.A., and González, P.A. (2019). Herpes Simplex Virus Type 1 Infection of the Central Nervous System: Insights Into Proposed Interrelationships With Neurodegenerative Disorders. Front. Cell. Neurosci., 13. 10.3389/fncel.2019.00046

[8]

Gutekunst "Isolation of pseudorabies virus from trigeminal ganglia of a latently infected sow" Am. J. Vet. Res. (1980) 10.2460/ajvr.1980.41.08.1315

[9]

Roizman, B. (1985). Molecular Biology of Pseudorabies Virus. The Herpesviruses: Volume 3, Springer. 10.1007/978-1-4613-2383-9

[10]

Molecular Biology of Pseudorabies Virus: Impact on Neurovirology and Veterinary Medicine

Lisa E. Pomeranz, Ashley E. Reynolds, Christoph J. Hengartner

Microbiology and Molecular Biology Reviews 2005 10.1128/mmbr.69.3.462-500.2005

[11]

Mahjoub "A 2.5-kilobase deletion containing a cluster of nine microRNAs in the latency-associated-transcript locus of the pseudorabies virus affects the host response of porcine trigeminal ganglia during established latency" J. Virol. (2015) 10.1128/jvi.02181-14

[12]

Anselmo, A., Flori, L., Jaffrezic, F., Rutigliano, T., Cecere, M., Cortes-Perez, N., Lefèvre, F., Rogel-Gaillard, C., and Giuffra, E. (2011). Co-Expression of Host and Viral MicroRNAs in Porcine Dendritic Cells Infected by the Pseudorabies Virus. PLoS ONE, 6. 10.1371/journal.pone.0017374

[13]

Perng "Virus-Induced Neuronal Apoptosis Blocked by the Herpes Simplex Virus Latency-Associated Transcript" Science (2000) 10.1126/science.287.5457.1500

[14]

MicroRNAs expressed by herpes simplex virus 1 during latent infection regulate viral mRNAs

Jennifer Lin Umbach, Martha F. Kramer, Igor Jurak et al.

Nature 2008 10.1038/nature07103

[15]

Shen "Two small RNAs encoded within the first 1.5 kilobases of the herpes simplex virus type 1 latency-associated transcript can inhibit productive infection and cooperate to inhibit apoptosis" J. Virol. (2009) 10.1128/jvi.00871-09

[16]

Divito "A Triple Entente: Virus, Neurons, and CD8+ T Cells Maintain HSV-1 Latency" Immunol. Res. (2006) 10.1385/ir:36:1:119

[17]

Harrison "Regulation of herpes simplex virus type 1 latency-reactivation cycle and ocular disease by cellular signaling pathways" Exp. Eye Res. (2022) 10.1016/j.exer.2022.109017

[18]

Cohen "Herpesvirus latency" J. Clin. Investig. (2020) 10.1172/jci136225

[19]

Yao "In vivo reactivation of latent herpes simplex virus 1 in mice can occur in the brain before occurring in the trigeminal ganglion" J. Virol. (2014) 10.1128/jvi.01616-14

[20]

Sivasubramanian "Herpes Simplex Virus Type 1 Preferentially Enhances Neuro-Inflammation and Senescence in Brainstem of Female Mice" J. Virol. (2022) 10.1128/jvi.01081-22

[21]

Herpes Simplex Virus-1 in the Brain: The Dark Side of a Sneaky Infection

Maria Elena Marcocci, Giorgia Napoletani, Virginia Protto et al.

Trends in Microbiology 2020 10.1016/j.tim.2020.03.003

[22]

Stahl "Herpes simplex virus encephalitis update" Curr. Opin. Infect. Dis. (2019) 10.1097/qco.0000000000000554

[23]

Bradshaw "Herpes Simplex Virus-1 Encephalitis in Adults: Pathophysiology, Diagnosis, and Management" Neurotherapeutics (2016) 10.1007/s13311-016-0433-7

[24]

Feng, S., Liu, Y., Zhou, Y., Shu, Z., Cheng, Z., Brenner, C., and Feng, P. (2023). Mechanistic insights into the role of herpes simplex virus 1 in Alzheimer’s disease. Front. Aging Neurosci., 15. 10.3389/fnagi.2023.1245904

[25]

Protto "Role of HSV-1 in Alzheimer’s disease pathogenesis: A challenge for novel preventive/therapeutic strategies" Curr. Opin. Pharmacol. (2022) 10.1016/j.coph.2022.102200

[26]

Mettenleiter "Aujeszky’s disease (pseudorabies) virus: The virus and molecular pathogenesis—State of the art, June 1999" Vet. Res. (2000)

[27]

Babic "Propagation of Pseudorabies Virus in the Nervous System of the Mouse after Intranasal Inoculation" Virology (1994) 10.1006/viro.1994.1576

[28]

Sehl, J., Hölper, J.E., Klupp, B.G., Baumbach, C., Teifke, J.P., and Mettenleiter, T.C. (2020). An improved animal model for herpesvirus encephalitis in humans. PLoS Pathog., 16. 10.1371/journal.ppat.1008445

[29]

Schwaiger "Clinical, neuropathological, and immunological short- and long-term feature of a mouse model mimicking human herpes virus encephalitis" Brain Pathol. (2022) 10.1111/bpa.13031

[30]

Armien "Chronic cortical and subcortical pathology with associated neurological deficits ensuing experimental herpes encephalitis" Brain Pathol. (2010) 10.1111/j.1750-3639.2009.00354.x

[31]

Asenbauer "Chronic active destructive herpes simplex encephalitis with recovery of viral DNA 12 years after disease onset" Neuropediatrics (1998) 10.1055/s-2007-973546

[32]

Armangue "Frequency, symptoms, risk factors, and outcomes of autoimmune encephalitis after herpes simplex encephalitis: A prospective observational study and retrospective analysis" Lancet Neurol. (2018) 10.1016/s1474-4422(18)30244-8

[33]

Osorio "A murine model of pseudorabies virus latency" Microb. Pathog. (1992) 10.1016/0882-4010(92)90064-u

[34]

Kaplan "A Comparison of Herpes Simplex and Pseudorabies Viruses" Virology (1959) 10.1016/0042-6822(59)90068-6

[35]

Gage "Whole animal perfusion fixation for rodents" J. Vis. Exp. (2012)

[36]

Rao "Histopathological evaluation of the nervous system in National Toxicology Program rodent studies: A modified approach" Toxicol. Pathol. (2011) 10.1177/0192623311401044

[37]

Paxinos, G., and Franklin, K.B.J. (2001). The Mouse Brain in Stereotaxic Coordinates, Academic Press. [2nd ed.].

[38]

RNAscope

Fay Wang, John Flanagan, Nan SU et al.

The Journal of Molecular Diagnostics 2012 10.1016/j.jmoldx.2011.08.002

[39]

Wernike "Molecular double-check strategy for the identification and characterization of Suid herpesvirus 1" J. Virol. Methods (2014) 10.1016/j.jviromet.2014.08.022

[40]

Olfactory and trigeminal routes of HSV-1 CNS infection with regional microglial heterogeneity

Christy S. Niemeyer, Laetitia Merle, Andrew N. Bubak et al.

Journal of Virology 2024 10.1128/jvi.00968-24

[41]

McFarland "Contrasting patterns of virus spread and neuropathology following microinjection of herpes simplex virus into the hippocampus or cerebellum of mice" J. Neurol. Sci. (1987) 10.1016/0022-510x(87)90233-4

[42]

Wu "Herpes simplex virus type 1 inoculation enhances hippocampal excitability and seizure susceptibility in mice" Eur. J. Neurosci. (2003) 10.1111/j.1460-9568.2003.03075.x

[43]

Chee, K., Razmara, A., Geller, A.S., Harris, W.B., Restrepo, D., Thompson, J.A., and Kramer, D.R. (2022). The role of the piriform cortex in temporal lobe epilepsy: A current literature review. Front. Neurol., 13. 10.3389/fneur.2022.1042887

[44]

Costa, B., and Vale, N. (2024). Virus-Induced Epilepsy vs. Epilepsy Patients Acquiring Viral Infection: Unravelling the Complex Relationship for Precision Treatment. Int. J. Mol. Sci., 25. 10.3390/ijms25073730

[45]

Wouk "Viral infections and their relationship to neurological disorders" Arch. Virol. (2021) 10.1007/s00705-021-04959-6

[46]

Held "Control of HSV-1 latency in human trigeminal ganglia--current overview" J. Neurovirol. (2011) 10.1007/s13365-011-0063-0

[47]

Geraghty "Entry of Alphaherpesviruses Mediated by Poliovirus Receptor-Related Protein 1 and Poliovirus Receptor" Science (1998) 10.1126/science.280.5369.1618

[48]

Lathe "Distribution of cellular HSV-1 receptor expression in human brain" J. Neurovirol. (2017) 10.1007/s13365-016-0504-x

[49]

Prandovszky "Spatiotemporal changes of the herpes simplex virus entry receptor nectin-1 in murine brain during postnatal development" J. Neurovirol. (2006) 10.1080/13550280600760594

[50]

Korff "Neurotropism of alphaherpesviruses is most prominent in the mesiotemporal, piriform and prefrontal cortices in mice" Neuroscience (2025) 10.1016/j.neuroscience.2025.08.024

Showing 50 of 63 references

Metrics

0

Citations

63

References

Details

Published: Apr 07, 2026
Vol/Issue: 15(4)
Pages: 395
License: View

Authors

V

Viktoria Korff

Department of Experimental Animal Facilities and Biorisk Management, Friedrich-Loeffler-Institut, 17493 Greifswald, Germany

I

Issam El-Debs

Department of Experimental Animal Facilities and Biorisk Management, Friedrich-Loeffler-Institut, 17493 Greifswald, Germany

B

Barbara G. Klupp

Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald, Germany

C

Conrad M. Freuling

Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald, Germany

J

Jens P. Teifke

Department of Experimental Animal Facilities and Biorisk Management, Friedrich-Loeffler-Institut, 17493 Greifswald, Germany

T

Thomas C. Mettenleiter

Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald, Germany

J

Julia Sehl-Ewert

Department of Experimental Animal Facilities and Biorisk Management, Friedrich-Loeffler-Institut, 17493 Greifswald, Germany

Funding

Deutsche Forschungsgemeinschaft Award: 466759708

Cite This Article

Viktoria Korff, Issam El-Debs, Barbara G. Klupp, et al. (2026). Regional and Temporal Patterns of Long-Term Pseudorabies Virus Detection and Neuropathology in the Murine CNS. Pathogens, 15(4), 395. https://doi.org/10.3390/pathogens15040395

Regional and Temporal Patterns of Long-Term Pseudorabies Virus Detection and Neuropathology in the Murine CNS

You May Also Like