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bioRxiv · 10.1101/2025.08.22.671689

Optimisation of lytic herpes simplex virus infection in human induced pluripotent stem cell derived cortical neurones

Nash, D. A.·Nicholson, A. S.·Barrow, H. G.·Connor, V.·Crump, C. M.·Deane, J. E.·Graham, S. C.
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Abstract

Herpes simplex virus (HSV)-1 infection of cortical neurones is a leading cause of encephalitis. While we have substantial knowledge about the molecular virology of HSV-1 lytic infection in cells of the periphery, like keratinocytes or fibroblasts, we know much less about infection of human neurones owing to the challenges of working with neuronal cell-based models. Here we demonstrate the use of a human induced pluripotent stem cell (iPSC)-derived cortical neurone model (i3Neurones) for HSV-1 infection. i3Neurones are highly scalable and can be rapidly and efficiently differentiated into an isogenic population of cortical glutamatergic neurones. We show that i3Neurones support the full HSV-1 lytic replication cycle. We present an optimised protocol for the infection of i3Neurones with HSV-1 that allows their synchronous infection at near-100% efficiency, and optimised fixation methods that preserves organelle and neurite structure for immunocytochemistry analysis. Our study highlights i3Neurones as a robust, scalable platform for microscopy and biochemical studies of HSV-1 and other neurotropic pathogens. Data summaryThe authors confirm all supporting data, code and protocols have been provided within the article or through supplementary data files.

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Nash, D. A., Nicholson, A. S., Barrow, H. G., Connor, V., Crump, C. M., Deane, J. E., Graham, S. C.. 2025-08-22. Optimisation of lytic herpes simplex virus infection in human induced pluripotent stem cell derived cortical neurones. https://doi.org/10.1101/2025.08.22.671689

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