title: An In Vitro HSV-1 Reactivation Model Containing Quiescently Infected PC12 Cells.
abstract: Advances in the understanding of the infection and reactivation process of herpes simplex type 1 (HSV-1) are generally gained by monolayer cultures or extensive and cost-intensive animal models. So far, no reliable in vitro skin model exists either to investigate the molecular mechanisms involved in controlling latency and virus reactivation or to test pharmaceuticals. Here we demonstrate the first in vitro HSV-1 reactivation model generated by using the human keratinocyte cell line HaCaT grown on a collagen substrate containing primary human fibroblasts. We integrated the unique feature of a quiescently infected neuronal cell line, the rat pheochromocytoma line PC12, within the dermal layer of the three-dimensional skin equivalent. Transmission electron microscopy, a cell-based TCID50 assay, and polymerase chain reaction analysis were used to verify cell latency. Thereby viral DNA could be detected, whereas extracellular as well as intracellular virus activity could not be found. Further, the infected PC12 cells show no spontaneous reactivation within the in vitro skin equivalent. In order to simulate a physiologically comparable HSV-1 infection, we achieved a specific and pointed reactivation of quiescently HSV-1 infected PC12 cells by UVB irradiation at 1000 mJ/cm(2).
authors.affiliation: Institute for Interfacial Engineering and Plasma Technology, University of Stuttgart , Stuttgart, Germany .
authors.forename: Ina
authors.initials: I
authors.lastname: Hogk
authors.forename: Michaela
authors.initials: M
authors.lastname: Kaufmann
authors.forename: Doris
authors.initials: D
authors.lastname: Finkelmeier
authors.forename: Steffen
authors.initials: S
authors.lastname: Rupp
authors.forename: Anke
authors.initials: A
authors.lastname: Burger-Kentischer
keywords: alternative testing; herpes simplex virus type 1; in vitro reactivation model; quiescent infection; virus reactivation
references.citation: J Virol. 1985 Aug;55(2):410-6
references.pmid: 2991566
references.citation: J Gen Virol. 1996 Oct;77 ( Pt 10):2583-90
references.pmid: 8887494
references.citation: Virology. 1997 Apr 14;230(2):236-43
references.pmid: 9143279
references.citation: Virology. 1983 May;127(1):159-67
references.pmid: 6305013
references.citation: J Gen Virol. 2002 Dec;83(Pt 12):2943-2950
references.pmid: 12466470
references.citation: J Neurovirol. 1999 Jun;5(3):258-67
references.pmid: 10414516
references.citation: J Gen Virol. 1999 Aug;80 ( Pt 8):2149-2155
references.pmid: 10466814
references.citation: J Neurovirol. 2000 Aug;6(4):341-9
references.pmid: 10951558
references.citation: J Infect Dis. 1992 Sep;166(3):500-6
references.pmid: 1323616
references.citation: Adv Drug Deliv Rev. 2002 Nov 1;54 Suppl 1:S19-30
references.pmid: 12460713
references.citation: J Neurovirol. 1995 Mar;1(1):19-29
references.pmid: 9222339
references.citation: Nature. 2007 Feb 22;445(7130):874-80
references.pmid: 17314974
references.citation: J Virol. 1994 Dec;68(12):8045-55
references.pmid: 7966594
references.citation: Proc Natl Acad Sci U S A. 1976 Jul;73(7):2424-8
references.pmid: 1065897
references.citation: Annu Rev Microbiol. 1987;41:543-71
references.pmid: 2825588
references.citation: J Virol. 1999 Mar;73(3):1767-73
references.pmid: 9971753
references.citation: J Gen Virol. 2002 Jul;83(Pt 7):1591-1600
references.pmid: 12075077
references.citation: J Virol Methods. 2001 Aug;96(2):107-26
references.pmid: 11445142
references.citation: Virology. 1996 Nov 1;225(1):72-81
references.pmid: 8918535
references.citation: J Gen Virol. 1994 Oct;75 ( Pt 10):2691-8
references.pmid: 7931155
references.citation: Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6529-33
references.pmid: 7517552
references.citation: Virology. 1987 Jun;158(2):265-75
references.pmid: 3035783
references.citation: Semin Cancer Biol. 2005 Oct;15(5):365-77
references.pmid: 15975824
references.citation: Gene Ther. 2005 Jun;12(11):891-901
references.pmid: 15908995
references.citation: Immunol Rev. 1996 Aug;152:157-73
references.pmid: 8930672
references.citation: Science. 1982 Sep 17;217(4565):1145-6
references.pmid: 6180477
references.citation: Altern Lab Anim. 2008 May;36(2):161-87
references.pmid: 18522484
references.citation: J Virol. 2001 Mar;75(6):2912-20
references.pmid: 11222716
references.citation: Tissue Eng Part A. 2010 Sep;16(9):2821-31
references.pmid: 20486787
references.citation: J Virol. 1997 Jul;71(7):5423-31
references.pmid: 9188614
references.citation: Physiol Rev. 2003 Apr;83(2):581-632
references.pmid: 12663867
references.citation: J Virol. 2000 Feb;74(4):1885-91
references.pmid: 10644361
references.citation: J Virol. 1996 Sep;70(9):6524-8
references.pmid: 8709294
references.citation: Curr Protoc Cell Biol. 2007 Sep;Chapter 15:Unit 15.12
references.pmid: 18228498
references.citation: Curr Protoc Microbiol. 2005 Oct;Chapter 14:Unit 14E.1
references.pmid: 18770556
references.citation: Virus Genes. 2006 Oct;33(2):163-7
references.pmid: 16972030
references.citation: GMS Krankenhhyg Interdiszip. 2008 Mar 11;3(1):Doc11
references.pmid: 20204113
references.citation: J Virol. 1999 May;73(5):4171-80
references.pmid: 10196313
references.citation: Invest Ophthalmol Vis Sci. 2010 Sep;51(9):4703-6
references.pmid: 20393107
references.citation: Brain. 1989 Oct;112 ( Pt 5):1277-94
references.pmid: 2804613
references.citation: Viral Immunol. 2005;18(3):466-73
references.pmid: 16212525
references.citation: Adv Drug Deliv Rev. 2011 Apr 30;63(4-5):352-66
references.pmid: 21241756
references.citation: Nature. 2008 Aug 7;454(7205):780-3
references.pmid: 18596690
references.citation: Arch Virol. 2008;153(7):1335-45
references.pmid: 18548318
references.citation: Microbiology (Reading). 2002 Feb;148(Pt 2):497-506
references.pmid: 11832513