title: An Immortalized Human Dorsal Root Ganglion Cell Line Provides a Novel Context To Study Herpes Simplex Virus 1 Latency and Reactivation.
abstract: A defining characteristic of alphaherpesviruses is the establishment of lifelong latency in host sensory ganglia with occasional reactivation causing recurrent lytic infections. As an alternative to rodent models, we explored the use of an immortalized cell line derived from human dorsal root ganglia. HD10.6 cells proliferate by virtue of a transduced tetracycline-regulated v-myconcogene. In the presence of doxycycline, HD10.6 cells mature to exhibit neuronal morphology and express sensory neuron-associated markers such as neurotrophin receptors TrkA, TrkB, TrkC, and RET and the sensory neurofilament peripherin. Infection of mature HD10.6 neurons by herpes simplex virus 1 (HSV-1) results in a delayed but productive infection. However, infection at a low multiplicity of infection (MOI) in the presence of acyclovir results in a quiescent infection resembling latency in which viral genomes are retained in a low number of neurons, viral gene expression is minimal, and infectious virus is not released. At least some of the quiescent viral genomes retain the capacity to reactivate, resulting in viral DNA replication and release of infectious virus. Reactivation can be induced by depletion of nerve growth factor; other commonly used reactivation stimuli have no significant effect.IMPORTANCEInfections by herpes simplex viruses (HSV) cause painful cold sores or genital lesions in many people; less often, they affect the eye or even the brain. After the initial infection, the virus remains inactive or latent in nerve cells that sense the region where that infection occurred. To learn how virus maintains and reactivates from latency, studies are done in neurons taken from rodents or in whole animals to preserve the full context of infection. However, some cellular mechanisms involved in HSV infection in rodents are different from those in humans. We describe the use of a human cell line that has the properties of a sensory neuron. HSV infection in these cultured cells shows the properties expected for a latent infection, including reactivation to produce newly infectious virus. Thus, we now have a cell culture model for latency that is derived from the normal host for this virus.
authors.affiliation: Center for Epigenetics, Van Andel Research Institute, Grand Rapids, Michigan, USA.
authors.forename: Nikki M
authors.initials: NM
authors.lastname: Thellman
authors.affiliation: Center for Epigenetics, Van Andel Research Institute, Grand Rapids, Michigan, USA.
authors.forename: Carolyn
authors.initials: C
authors.lastname: Botting
authors.affiliation: Bioinformatics and Biostatistics Core, Van Andel Research Institute, Grand Rapids, Michigan, USA.
authors.forename: Zachary
authors.initials: Z
authors.lastname: Madaj
authors.affiliation: Center for Epigenetics, Van Andel Research Institute, Grand Rapids, Michigan, USA steve.triezenberg@vai.edu.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Triezenberg
mesh_terms: D000212:Acyclovir / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology; D018929:Cell Culture Techniques; D002460:Cell Line; D004318:Doxycycline / Q000494:pharmacology; D005727:Ganglia, Spinal / Q000187:drug effects / Q000821:virology*; D015967:Gene Expression Regulation, Viral; D006561:Herpes Simplex / Q000821:virology; D018259:Herpesvirus 1, Human / Q000502:physiology*; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics; D020932:Nerve Growth Factor / Q000494:pharmacology; D064531:Peripherins / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D051096:Proto-Oncogene Proteins c-ret / Q000235:genetics; D020917:Receptor, trkA / Q000235:genetics; D020813:Receptor, trkB; D020812:Receptor, trkC / Q000235:genetics; D011984:Sensory Receptor Cells / Q000502:physiology / Q000821:virology*; D014775:Virus Activation; D017735:Virus Latency; D057074:Virus Release / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D008562:Membrane Glycoproteins; C000611485:NTRK1 protein, human; C578994:PRPH protein, human; D064531:Peripherins; D020932:Nerve Growth Factor; D011505:Protein-Tyrosine Kinases; D051096:Proto-Oncogene Proteins c-ret; C099282:RET protein, human; D020917:Receptor, trkA; D020813:Receptor, trkB; D020812:Receptor, trkC; C000591259:tropomyosin-related kinase-B, human; D004318:Doxycycline; D000212:Acyclovir
keywords: HSV-1; quiescent infection; sensory neurons