Differentiated Human SH-SY5Y Cells Provide a Reductionist Model of Herpes Simplex Virus 1 Neurotropism.

نویسندگان

  • Mackenzie M Shipley
  • Colleen A Mangold
  • Chad V Kuny
  • Moriah L Szpara
چکیده

Neuron-virus interactions that occur during herpes simplex virus (HSV) infection are not fully understood. Neurons are the site of lifelong latency and are a crucial target for long-term suppressive therapy or viral clearance. A reproducible neuronal model of human origin would facilitate studies of HSV and other neurotropic viruses. Current neuronal models in the herpesvirus field vary widely and have caveats, including incomplete differentiation, nonhuman origins, or the use of dividing cells that have neuropotential but lack neuronal morphology. In this study, we used a robust approach to differentiate human SH-SY5Y neuroblastoma cells over 2.5 weeks, producing a uniform population of mature human neuronal cells. We demonstrate that terminally differentiated SH-SY5Y cells have neuronal morphology and express proteins with subcellular localization indicative of mature neurons. These neuronal cells are able to support a productive HSV-1 infection, with kinetics and overall titers similar to those seen in undifferentiated SH-SY5Y cells and the related SK-N-SH cell line. However, terminally differentiated, neuronal SH-SY5Y cells release significantly less extracellular HSV-1 by 24 h postinfection (hpi), suggesting a unique neuronal response to viral infection. With this model, we are able to distinguish differences in neuronal spread between two strains of HSV-1. We also show expression of the antiviral protein cyclic GMP-AMP synthase (cGAS) in neuronal SH-SY5Y cells, which is the first demonstration of the presence of this protein in nonepithelial cells. These data provide a model for studying neuron-virus interactions at the single-cell level as well as via bulk biochemistry and will be advantageous for the study of neurotropic viruses in vitroIMPORTANCE Herpes simplex virus (HSV) affects millions of people worldwide, causing painful oral and genital lesions, in addition to a multitude of more severe symptoms such as eye disease, neonatal infection, and, in rare cases, encephalitis. Presently, there is no cure available to treat those infected or prevent future transmission. Due to the ability of HSV to cause a persistent, lifelong infection in the peripheral nervous system, the virus remains within the host for life. To better understand the basis of virus-neuron interactions that allow HSV to persist within the host peripheral nervous system, improved neuronal models are required. Here we describe a cost-effective and scalable human neuronal model system that can be used to study many neurotropic viruses, such as HSV, Zika virus, dengue virus, and rabies virus.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Establishing cell model of herpes simplex virus type 2 latent infection and reactivation in SH-SY5Y cells

Objective: To establish cell model of herpes simplex virus type 2 latent infection and reactivation in SHSY5Y cells. Methods: Effects of ACV on SH-SY5Y cells were observed after cells were treated with ACV concentrations of 20, 40, 60, 80, 100, 120, or 140 μmol/L, and also cells morphology was observed under phase contrast microscope after HSV-2 was inoculated into SH-SY5Y cells using MOI of 0....

متن کامل

Up-Regulation of Integrinsn α2β1 and α3β1 Expression in Human Foreskin Fibroblast Cells after In-Vitro Infection with Herpes Simplex Virus Type 1

The interaction of Herpes Simplex Virus type 1 (HSV-1) with human fetal foreskin fibroblast (HFFF) cell was studied using a recent isolate of HSV-1 which was propagated in Hep-2 cells. HFFF cells were challenged with HSV-1 with a multiplicity of infection (MOI) of 1 virus/cell for 24 hours. Flow cytometric analysis demonstrated that HSV-1 challenged HFFF cells expressed increased levels of α2β1...

متن کامل

Effects of Morphine on Replication of Herpes Simplex Virus Type 1&2

Background and Aims: Several drugs are being used in treatment of HSV infection in human but still introducing an effective safe drug is desirable. Methods: We investigated the inhibitory effect of morphine on replication of HSV in vitro. Results: The results indicated that a concentration of up to 200 ug/ml morphine had a limited effect on Vero cell viability. At this concentration the growt...

متن کامل

Effects of Sodium Valproate on the Replication of Herpes Simplex Virus Type 1: An in Vitro Study

Background: Sodium valproate, an anticonvulsant drug, is reported to stimulate Human Immunodeficiency Virus type 1 and Human cytomegalovirus replication. Since epileptic patients undergoing sodium valproate therapy may suffer from various virus infections, the effect of this drug on replication of viruses especially those affecting neuronal tissues such as Herpes simplex virus type 1 is worthy ...

متن کامل

Replication Characteristics of Herpes Simplex Virus Type-1 (HSV-1) Recombinants in 3 Types of Tissue Cultures

A complication in the analysis of the role of ICP34.5 gene in the herpes simplex virus type-1 (HSV-1) lifecycle is the presence of overlapping antisense gene, open reading frame P (ORF P), which is also deleted in HSV-1 ICP34.5 negative mutants. A HSV-1 wild type strain (17+) ICP34.5/ORF P deletion mutant (1716) is totally avirulent in animal models and impaired in a number of in vitro function...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Journal of virology

دوره 91 23  شماره 

صفحات  -

تاریخ انتشار 2017