Abstract
We describe a primary neuronal culture system suitable for molecular characterization of herpes simplex virus type 1 (HSV-1) infection, latency, and reactivation. While several alternative models are available, including infections of live animal and explanted ganglia, these are complicated by the presence of multiple cell types, including immune cells, and difficulties in manipulating the neuronal environment. The highly pure neuron culture system described here can be readily manipulated and is ideal for molecular studies that focus exclusively on the relationship between the virus and host neuron, the fundamental unit of latency. As such it allows for detailed investigations of both viral and neuronal factors involved in the establishment and maintenance of HSV-1 latency and in viral reactivation induced by defined stimuli.
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References
Wilson AC, Mohr I (2012) A cultured affair: HSV latency and reactivation in neurons. Trends Microbiol 20:604–611
Wagner EK, Bloom DC (1997) Experimental investigation of herpes simplex virus latency. Clin Microbiol Rev 10:419–443
Kobayashi M, Wilson AC, Chao MV et al (2012) Control of viral latency in neurons by axonal mTOR signaling and the 4E-BP translation repressor. Genes Dev 26:1527–1532
Camarena V, Kobayashi M, Kim JY et al (2010) Nature and duration of growth factor signaling through receptor tyrosine kinases regulates HSV-1 latency in neurons. Cell Host Microbe 8:320–330
Wilcox CL, Smith RL, Freed CR et al (1990) Nerve growth factor-dependence of herpes simplex virus latency in peripheral sympathetic and sensory neurons in vitro. J Neurosci 10: 1268–1275
Wilcox CL, Johnson EM (1988) Characterization of nerve growth factor-dependent herpes simplex virus latency in neurons in vitro. J Virol 62:393–399
Roehm PC, Camarena V, Nayak S et al (2011) Cultured vestibular ganglion neurons demonstrate latent HSV1 reactivation. Laryngoscope 121:2268–2275
Kuhn MA, Nayak S, Camarena V et al (2012) A cell culture model of facial palsy resulting from reactivation of latent herpes simplex type 1. Otol Neurotol 33:87–92
Warren KG, Brown SM, Wroblewska Z et al (1978) Isolation of latent herpes simplex virus from the superior cervical and vagus ganglions of human beings. N Engl J Med 298: 1068–1069
Price RW, Katz BJ, Notkins AL (1975) Latent infection of the peripheral ANS with herpes simplex virus. Nature 257:686–688
Kim JY, Mandarino A, Chao MV et al (2012) Transient reversal of episome silencing precedes VP16-dependent transcription during reactivation of latent HSV-1 in neurons. PLoS Pathog 8:e1002540
Harlow E, Lane D (eds) (1988) Antibodies. A laboratory manual. Cold Spring Harbor Press, Cold Spring Harbor, NY
Sambrook J, Fritsch EF, Maniatis T (eds) (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Press, Cold Spring Harbor, NY
Acknowledgement
We thank our colleagues Moses Chao and Ed Ziff and the members of their laboratories for teaching us so much about the isolation and culture of neurons and for graciously hosting us in the aftermath of hurricane Sandy. This work was supported by grants from the National Institutes of Health (AI073898, GM61139, S10RR017970, T32AI007647, and T32AI07180), the Vilcek Foundation, and a collaborative project grant from the New York University Langone Medical Center.
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Kim, J.Y., Shiflett, L.A., Linderman, J.A., Mohr, I., Wilson, A.C. (2014). Using Homogeneous Primary Neuron Cultures to Study Fundamental Aspects of HSV-1 Latency and Reactivation. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus. Methods in Molecular Biology, vol 1144. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0428-0_11
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DOI: https://doi.org/10.1007/978-1-4939-0428-0_11
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-0428-0
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