This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wang, K.
Right arrow Articles by Straus, S. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wang, K.
Right arrow Articles by Straus, S. E.

 Previous Article  |  Next Article 

Journal of Virology, November 2005, p. 14079-14087, Vol. 79, No. 22
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.22.14079-14087.2005

Laser-Capture Microdissection: Refining Estimates of the Quantity and Distribution of Latent Herpes Simplex Virus 1 and Varicella-Zoster Virus DNA in Human Trigeminal Ganglia at the Single-Cell Level

Kening Wang,* Tsz Y. Lau, Melissa Morales, Erik K. Mont,{dagger} and Stephen E. Straus

Medical Virology Section, Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland

Received 9 June 2005/ Accepted 15 August 2005

There remains uncertainty and some controversy about the percentages and types of cells in human sensory nerve ganglia that harbor latent herpes simplex virus 1 (HSV-1) and varicella-zoster virus (VZV) DNA. We developed and validated laser-capture microdissection and real-time PCR (LCM/PCR) assays for the presence and copy numbers of HSV-1 gG and VZV gene 62 sequences in single cells recovered from sections of human trigeminal ganglia (TG) obtained at autopsy. Among 970 individual sensory neurons from five subjects, 2.0 to 10.5% were positive for HSV-1 DNA, with a median of 11.3 copies/positive cell, compared with 0.2 to 1.5% of neurons found to be positive by in situ hybridization (ISH) for HSV-1 latency-associated transcripts (LAT), the classical surrogate marker for HSV latency. This indicates a more pervasive latent HSV-1 infection of human TG neurons than originally thought. Combined ISH/LCM/PCR assays revealed that the majority of the latently infected neurons do not accumulate LAT to detectable levels. We detected VZV DNA in 1.0 to 6.9% of individual neurons from 10 subjects. Of the total 1,722 neurons tested, 4.1% were VZV DNA positive, with a median of 6.9 viral genomes/positive cell. After removal by LCM of all visible neurons on a slide, all surrounding nonneuronal cells were harvested and assayed: 21 copies of HSV-1 DNA were detected in ~5,200 nonneuronal cells, while nine VZV genomes were detected in ~14,200 nonneuronal cells. These data indicate that both HSV-1 and VZV DNAs persist in human TG primarily, if not exclusively, in a moderate percentage of neuronal cells.

* Corresponding author. Mailing address: Medical Virology Section, Laboratory of Clinical Infectious Diseases, NIAID/NIH, Building 10, Room 11N-228, 10 Center Drive, Bethesda, MD 20892. Phone: (301) 496-7895. Fax: (301) 496-7383. E-mail: kwang{at}niaid.nih.gov.

{dagger} Present address: Miami-Dade County Medical Examiner Department, Miami, FL 33136.

Journal of Virology, November 2005, p. 14079-14087, Vol. 79, No. 22
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.22.14079-14087.2005




This article has been cited by other articles:

  • Proenca, J. T., Coleman, H. M., Connor, V., Winton, D. J., Efstathiou, S. (2008). A historical analysis of herpes simplex virus promoter activation in vivo reveals distinct populations of latently infected neurones. J. Gen. Virol. 89: 2965-2974 [Abstract] [Full Text]  
  • Reichelt, M., Zerboni, L., Arvin, A. M. (2008). Mechanisms of Varicella-Zoster Virus Neuropathogenesis in Human Dorsal Root Ganglia. J. Virol. 82: 3971-3983 [Abstract] [Full Text]  
  • Zerboni, L., Reichelt, M., Jones, C. D., Zehnder, J. L., Ito, H., Arvin, A. M. (2007). From the Cover: Aberrant infection and persistence of varicella-zoster virus in human dorsal root ganglia in vivo in the absence of glycoprotein I. Proc. Natl. Acad. Sci. USA 104: 14086-14091 [Abstract] [Full Text]  
  • Wang, K., Mahalingam, G., Hoover, S. E., Mont, E. K., Holland, S. M., Cohen, J. I., Straus, S. E. (2007). Diverse Herpes Simplex Virus Type 1 Thymidine Kinase Mutants in Individual Human Neurons and Ganglia. J. Virol. 81: 6817-6826 [Abstract] [Full Text]  
  • Thompson, R. L., Sawtell, N. M. (2006). Evidence that the Herpes Simplex Virus Type 1 ICP0 Protein Does Not Initiate Reactivation from Latency In Vivo. J. Virol. 80: 10919-10930 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»