{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2025,12,15]],"date-time":"2025-12-15T16:04:01Z","timestamp":1765814641333,"version":"3.48.0"},"posted":{"date-parts":[[2025,12,12]]},"group-title":"Microbiology","reference-count":68,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,12,12]],"date-time":"2025-12-12T00:00:00Z","timestamp":1765497600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,12,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Varicella-zoster virus (VZV) is a ubiquitous human neurotropic alphaherpesvirus that establishes lifelong latency in sensory ganglionic neurons. Subsequent viral reactivation causes herpes zoster, a morbid disease often complicated by neuropathic pain. Mechanisms underlying VZV latency and reactivation are not understood, mostly due to the lack of permissive animal models and challenges of current\n                  <jats:italic>in vitro<\/jats:italic>\n                  latency modelling. Here, we evaluated HD10.6 cells, a simplified and easily expandable human sensory neuron line to model VZV latency and reactivation. Mature HD10.6 (mHD10.6) differentiated neurons supported productive VZV infection, viral DNA replication, production of infectious progeny, and viral spread in cultures. VZV infection was associated with limited cytopathic effects and ultrastructural changes. Infecting mHD10.6 neurons with cell-free VZV in the presence of antivirals resulted in a quiescent-persistent state, characterized by persistent VZV genomes with restricted VZV gene expression and absence of infectious virus. Importantly, VZV could be reactivated by treatment with capsaicin, as evidenced by increased lytic viral transcription and virus spread. In conclusion, this study establishes human HD10.6 neurons as a novel and scalable\n                  <jats:italic>in vitro<\/jats:italic>\n                  model for studying VZV latency and reactivation to identify virus and host factors governing latency that may serve as therapeutic targets to restrict VZV reactivation.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Importance<\/jats:title>\n                  <jats:p>Most individuals carry latent varicella-zoster virus (VZV) in their dorsal root ganglia (DRG), which can reactivate to cause shingles and chronic pain. The mechanisms by which VZV establishes latency and triggers of reactivation are incompletely understood. Current platforms for the study of VZV latency do not easily support functional experiments (ganglia) or are difficult to expand and complicated by mixed neuronal populations (stem cell-derived neurons). Here, we demonstrate that matured HD10.6 (mHD10.6) cells derived from immortalized human DRG-derived neurons provide a clonal, scalable, and easy-to-expand platform for studying lytic, latent, and reactivated VZV infection in sensory neurons. We propose that the HD10.6 platform could provide the basis to conduct studies on the viral latent state that have hitherto not been possible.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2025.12.12.693952","type":"posted-content","created":{"date-parts":[[2025,12,13]],"date-time":"2025-12-13T05:05:11Z","timestamp":1765602311000},"source":"Crossref","is-referenced-by-count":0,"title":["A human sensory neuron model for varicella-zoster virus latency and reactivation\n                  <i>in vitro<\/i>"],"prefix":"10.64898","author":[{"given":"Alexander C.","family":"Havelaar","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph S.","family":"Flot","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saori","family":"Fukuda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anja W.M.","family":"de Jong","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R.I.","family":"Koning","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amber","family":"Schotting","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sem","family":"van \u2018t Geloof","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomohiko","family":"Sadaoka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2465-2674","authenticated-orcid":false,"given":"Georges M.G.M.","family":"Verjans","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1901-9970","authenticated-orcid":false,"given":"Paul R.","family":"Kinchington","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8393-296X","authenticated-orcid":false,"given":"Werner J.D.","family":"Ouwendijk","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025121508000730000_2025.12.12.693952v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2009.06.036"},{"key":"2025121508000730000_2025.12.12.693952v1.2","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1002\/ana.410020313","article-title":"Attempts to recover varicella virus from ganglia","volume":"2","year":"1977","journal-title":"Ann. Neurol"},{"key":"2025121508000730000_2025.12.12.693952v1.3","first-page":"478","article-title":"Varicella-zoster virus DNA in human sensory ganglia","volume":"307","year":"1983","journal-title":"Nature"},{"key":"2025121508000730000_2025.12.12.693952v1.4","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jiy407"},{"key":"2025121508000730000_2025.12.12.693952v1.5","doi-asserted-by":"crossref","unstructured":"Gershon, A. A. et al. Varicella zoster virus infection. Nat. Rev. Dis. Primer 1, 15016 (2015).","DOI":"10.1038\/nrdp.2015.16"},{"key":"2025121508000730000_2025.12.12.693952v1.6","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.69.12.8151-8154.1995"},{"key":"2025121508000730000_2025.12.12.693952v1.7","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.8.4658"},{"key":"2025121508000730000_2025.12.12.693952v1.8","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.77.12.6979-6987.2003"},{"key":"2025121508000730000_2025.12.12.693952v1.9","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.79.22.14079-14087.2005"},{"key":"2025121508000730000_2025.12.12.693952v1.10","doi-asserted-by":"crossref","unstructured":"Depledge, D. P. , Sadaoka, T. & Ouwendijk, W. J. D . Molecular Aspects of Varicella-Zoster Virus Latency. Viruses 10, 349 (2018).","DOI":"10.3390\/v10070349"},{"key":"2025121508000730000_2025.12.12.693952v1.11","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.80.10.4921-4926.2006"},{"key":"2025121508000730000_2025.12.12.693952v1.12","doi-asserted-by":"crossref","unstructured":"Depledge, D. P. et al. A spliced latency-associated VZV transcript maps antisense to the viral transactivator gene 61. Nat. Commun. 9, (2018).","DOI":"10.1038\/s41467-018-03569-2"},{"key":"2025121508000730000_2025.12.12.693952v1.13","doi-asserted-by":"crossref","unstructured":"Braspenning, S. E. et al. Decoding the Architecture of the Varicella-Zoster Virus Transcriptome. mBio 11, (2020).","DOI":"10.1128\/mBio.01568-20"},{"key":"2025121508000730000_2025.12.12.693952v1.14","doi-asserted-by":"crossref","unstructured":"Ouwendijk, W. J. D. et al. Varicella-zoster virus VLT-ORF63 fusion transcript induces broad viral gene expression during reactivation from neuronal latency. Nat. Commun. 11, (2020).","DOI":"10.1038\/s41467-020-20031-4"},{"key":"2025121508000730000_2025.12.12.693952v1.15","doi-asserted-by":"crossref","unstructured":"Baird, N. L. , Zhu, S. , Pearce, C. M. & Viejo-Borbolla, A . Current In Vitro Models to Study Varicella Zoster Virus Latency and Reactivation. Viruses 11, 103 (2019).","DOI":"10.3390\/v11020103"},{"key":"2025121508000730000_2025.12.12.693952v1.16","doi-asserted-by":"crossref","unstructured":"Laemmle, L. , Goldstein, R. S. & Kinchington, P. R . Modeling Varicella Zoster Virus Persistence and Reactivation \u2013 Closer to Resolving a Perplexing Persistent State. Front. Microbiol. 10, (2019).","DOI":"10.3389\/fmicb.2019.01634"},{"key":"2025121508000730000_2025.12.12.693952v1.17","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1004885"},{"key":"2025121508000730000_2025.12.12.693952v1.18","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1522575113"},{"key":"2025121508000730000_2025.12.12.693952v1.19","doi-asserted-by":"crossref","unstructured":"Kurapati, S. et al. Role of the JNK Pathway in Varicella-Zoster Virus Lytic Infection and Reactivation. J. Virol. 91, (2017).","DOI":"10.1128\/JVI.00640-17"},{"key":"2025121508000730000_2025.12.12.693952v1.20","doi-asserted-by":"crossref","unstructured":"Sadaoka, T. , et al. Human stem cell derived sensory neurons are positioned to support varicella zoster virus latency. (2020).","DOI":"10.1101\/2020.01.24.919290"},{"key":"2025121508000730000_2025.12.12.693952v1.21","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0013989"},{"key":"2025121508000730000_2025.12.12.693952v1.22","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00515-11"},{"key":"2025121508000730000_2025.12.12.693952v1.23","doi-asserted-by":"crossref","first-page":"e1012367","DOI":"10.1371\/journal.ppat.1012367","article-title":"Repression of varicella zoster virus gene expression during quiescent infection in the absence of detectable histone deposition","volume":"21","year":"2025","journal-title":"PLOS Pathog"},{"key":"2025121508000730000_2025.12.12.693952v1.24","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.19-13-05420.1999"},{"key":"2025121508000730000_2025.12.12.693952v1.25","doi-asserted-by":"crossref","unstructured":"Thellman, N. M. , Botting, C. , Madaj, Z. & Triezenberg, S. J . An Immortalized Human Dorsal Root Ganglion Cell Line Provides a Novel Context To Study Herpes Simplex Virus 1 Latency and Reactivation. J. Virol. 91, JVI.00080-17 (2017).","DOI":"10.1128\/JVI.00080-17"},{"key":"2025121508000730000_2025.12.12.693952v1.26","doi-asserted-by":"crossref","unstructured":"Haberberger, R. V. , Barry, C. & Matusica, D . Immortalized Dorsal Root Ganglion Neuron Cell Lines. Front. Cell. Neurosci. 14, (2020).","DOI":"10.3389\/fncel.2020.00184"},{"key":"2025121508000730000_2025.12.12.693952v1.27","doi-asserted-by":"crossref","unstructured":"Chen, Y.-C. , Li, H. , Martin-Caraballo, M. & Hsia, S. V . Establishing a Herpesvirus Quiescent Infection in Differentiated Human Dorsal Root Ganglion Neuronal Cell Line Mediated by Micro-RNA Overexpression. Pathogens 11, 803 (2022).","DOI":"10.3390\/pathogens11070803"},{"key":"2025121508000730000_2025.12.12.693952v1.28","doi-asserted-by":"crossref","unstructured":"Zhang, Q. , Martin-Caraballo, M. & Hsia, S. V . Modulation of Voltage-Gated Sodium Channel Activity in Human Dorsal Root Ganglion Neurons by Herpesvirus Quiescent Infection. J. Virol. 94, (2020).","DOI":"10.1128\/JVI.01823-19"},{"key":"2025121508000730000_2025.12.12.693952v1.29","doi-asserted-by":"crossref","unstructured":"Sadaoka, T. et al. A Variant Allele in Varicella-Zoster Virus Glycoprotein B Selected during Production of the Varicella Vaccine Contributes to Its Attenuation. mBio 13, (2022).","DOI":"10.1128\/mbio.01864-22"},{"key":"2025121508000730000_2025.12.12.693952v1.30","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01183-07"},{"key":"2025121508000730000_2025.12.12.693952v1.31","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02245-13"},{"key":"2025121508000730000_2025.12.12.693952v1.32","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.77.21.11718-11732.2003"},{"key":"2025121508000730000_2025.12.12.693952v1.33","first-page":"e01108","article-title":"Human Embryonic Stem Cell-Derived Neurons Are Highly Permissive for Varicella-Zoster Virus Lytic Infection","volume":"92","year":"2018","journal-title":"J. Virol"},{"key":"2025121508000730000_2025.12.12.693952v1.34","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.15.4.554"},{"key":"2025121508000730000_2025.12.12.693952v1.35","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.29.4.589"},{"key":"2025121508000730000_2025.12.12.693952v1.36","doi-asserted-by":"publisher","DOI":"10.2165\/00003495-200464180-00011"},{"key":"2025121508000730000_2025.12.12.693952v1.37","first-page":"359","article-title":"Brivudine: a herpes virostatic with rapid antiviral activity and once-daily dosing","volume":"39","year":"2003","journal-title":"Drugs Today Barc. Spain 1998"},{"key":"2025121508000730000_2025.12.12.693952v1.38","doi-asserted-by":"publisher","DOI":"10.1016\/S0166-3542(03)00065-2"},{"key":"2025121508000730000_2025.12.12.693952v1.39","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.61.7.2311-2315.1987"},{"key":"2025121508000730000_2025.12.12.693952v1.40","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.62.2.393-399.1988"},{"key":"2025121508000730000_2025.12.12.693952v1.41","first-page":"573","article-title":"Helicase-primase inhibitor amenamevir for herpesvirus infection: Towards practical application for treating herpes zoster","volume":"53","year":"2017","journal-title":"Drugs Today Barc. Spain 1998"},{"key":"2025121508000730000_2025.12.12.693952v1.42","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.antiviral.2016.12.008","article-title":"Profile of anti-herpetic action of ASP2151 (amenamevir) as a helicase-primase inhibitor","volume":"139","year":"2017","journal-title":"Antiviral Res"},{"key":"2025121508000730000_2025.12.12.693952v1.43","doi-asserted-by":"publisher","DOI":"10.3390\/v13081547"},{"key":"2025121508000730000_2025.12.12.693952v1.44","first-page":"330","article-title":"Sensory neuron-specific actions of capsaicin: mechanisms and applications","volume":"11","year":"1990","journal-title":"Trends Pharmacol. Sci"},{"key":"2025121508000730000_2025.12.12.693952v1.45","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2022.03.003"},{"key":"2025121508000730000_2025.12.12.693952v1.46","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1004989"},{"key":"2025121508000730000_2025.12.12.693952v1.47","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00445-11"},{"key":"2025121508000730000_2025.12.12.693952v1.48","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01506-13"},{"key":"2025121508000730000_2025.12.12.693952v1.49","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.75.19.9483-9492.2001"},{"key":"2025121508000730000_2025.12.12.693952v1.50","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1004173"},{"key":"2025121508000730000_2025.12.12.693952v1.51","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.77.23.12852-12864.2003"},{"key":"2025121508000730000_2025.12.12.693952v1.52","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2012.08.005"},{"key":"2025121508000730000_2025.12.12.693952v1.53","doi-asserted-by":"crossref","unstructured":"Baranovskiy, A. G. et al. Structural basis of herpesvirus helicase-primase inhibition by pritelivir and amenamevir. Sci. Adv. 11, eadz1989 (2025).","DOI":"10.1126\/sciadv.adz1989"},{"key":"2025121508000730000_2025.12.12.693952v1.54","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-025-02168-4"},{"key":"2025121508000730000_2025.12.12.693952v1.55","doi-asserted-by":"publisher","DOI":"10.1016\/0002-9343(82)90055-9"},{"key":"2025121508000730000_2025.12.12.693952v1.56","doi-asserted-by":"crossref","first-page":"e58037","DOI":"10.7554\/eLife.58037","article-title":"Neuronal hyperexcitability is a DLK-dependent trigger of herpes simplex virus reactivation that can be induced by IL-1","volume":"9","year":"2020","journal-title":"eLife"},{"key":"2025121508000730000_2025.12.12.693952v1.57","doi-asserted-by":"publisher","DOI":"10.1080\/13550280590952781"},{"key":"2025121508000730000_2025.12.12.693952v1.58","doi-asserted-by":"crossref","first-page":"1592","DOI":"10.1093\/infdis\/jiy358","article-title":"High Viral Diversity and Mixed Infections in Cerebral Spinal Fluid From Cases of Varicella Zoster Virus Encephalitis","volume":"218","year":"2018","journal-title":"J. Infect. Dis"},{"key":"2025121508000730000_2025.12.12.693952v1.59","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msu406"},{"key":"2025121508000730000_2025.12.12.693952v1.60","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6976.2011.00320.x"},{"key":"2025121508000730000_2025.12.12.693952v1.61","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00204-11"},{"key":"2025121508000730000_2025.12.12.693952v1.62","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2004.03.062"},{"key":"2025121508000730000_2025.12.12.693952v1.63","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01838-09"},{"key":"2025121508000730000_2025.12.12.693952v1.64","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02245-13"},{"key":"2025121508000730000_2025.12.12.693952v1.65","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2014.05.027"},{"key":"2025121508000730000_2025.12.12.693952v1.66","doi-asserted-by":"publisher","DOI":"10.1111\/j.2044-8295.1908.tb00176.x"},{"key":"2025121508000730000_2025.12.12.693952v1.67","doi-asserted-by":"publisher","DOI":"10.1007\/s12250-020-00230-5"},{"key":"2025121508000730000_2025.12.12.693952v1.68","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.201201140"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2025.12.12.693952","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,15]],"date-time":"2025-12-15T16:00:22Z","timestamp":1765814422000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2025.12.12.693952"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,12]]},"references-count":68,"URL":"https:\/\/doi.org\/10.64898\/2025.12.12.693952","relation":{},"subject":[],"published":{"date-parts":[[2025,12,12]]},"subtype":"preprint"}}