Reactivated and latent varicella-zoster virus in human dorsal root ganglia.

Abstract

Ganglia obtained at autopsy were examined by in situ hybridization from one patient with zoster (also called herpes zoster or shingles), two varicella-zoster virus (VZV)-seropositive patients without clinical evidence of zoster, one VZV-seronegative child, and one fetus. Ganglia positive for VZV had a hybridization signal in both neuronal and nonneuronal satellite cells. Ganglia obtained from the fetus and from the seronegative infant were consistently negative for VZV. Two striking observations were evident regarding the presence of VZV DNA in ganglia obtained from the individual with zoster at the time of death. First, ganglia innervating the sites of reactivation and ganglia innervating adjacent sites yielded strongly positive signals in neurons and satellite cells, whereas ganglia from distant sites were rarely positive. Second, VZV DNA was found in both the nuclei and the cytoplasm of neurons innervating areas of zoster. However, in neurons innervating zoster-free areas, VZV DNA was found only in the nucleus of neurons and their supporting satellite cells. Im Understanding the cellular and viral factors governing VZV latency and reactivation is essential to achieve this goal. Determining the location of persistent and reactivated virus is essential for defining the molecular basis of VZV pathogenesis.Electron microscopy has demonstrated VZV in sensory ganglia of patients with zoster (5, 6), and polymerase chain reaction (PCR) and Southern blot hybridization have detected VZV DNA in latently infected dorsal root ganglia (7,8). Studies utilizing in situ hybridization methods have identified several different VZV transcripts in affected ganglia (7,(9)(10)(11)(12)(13).These studies have led to controversy as to the nature of the cell harboring the latent VZV genome. VZV transcripts have been localized exclusively in neurons (10, 13, 14) or in nonneuronal satellite cells (9, 12) depending on the in situ hybridization technique used.In the current study, the cellular and intracellular location of the VZV genome in latently infected human dorsal root ganglia and in ganglia with clinical evidence of reactivated virus was determined by in situ hybridization. Evidence for virus replication in affected ganglia was provided by polymerase chain reaction (PCR) and immunohistochemistry. These results demonstrate that latent VZV resides in both neurons and their supporting satellite cells and that latent virus can reactivate in neurons. MATERIALS AND METHODSTissue Specimens. Dorsal root ganglia from two VZVseropositive patients without clinical evidence of zoster, one VZV-seronegative child and one fetus without a maternal history of varicella, were obtained at autopsy. Ganglia from an 85-year-old man with malignant glioma of the thoracic spinal cord who had a zosteriform rash in the distribution of the right Ti1, T12, and Li sensory nerves at the time of death were also obtained. Prior to developing zoster, the patient had radiation therapy to the spinal cord at the levels of T4 through Li. At autopsy, he had a ve...

Anatomical mapping of human herpesvirus reservoirs of infection

“…[9][10][11][13][14][15][16] Dissemination may lead to widespread organ involvement, including the eye, brain, lung, myocardium, and liver. Primary VZV infection, most often acquired in early childhood, is characterized by fever and a diffuse maculopapular rash.…”

The DNA Element Controlling Expression of the Varicella-Zoster Virus Open Reading Frame 28 and 29 Genes Consists of Two Divergent Unidirectional Promoters Which Have a Common USF Site

“…Transcription initiation from the ORF28 promoter was primed with a 26-base oligonucleotide (5Ј-AACTTT CGAAGTCATGGTGGCTAGGC-3Ј) complementary to a portion of the RL located 10 nucleotides upstream of the 5Ј end of the ORF 28/29 regulatory element. Both oligonucleotides were commercially synthesized (IDT, Coralville, Iowa), polyacrylamide gel electrophoresis (PAGE) purified, and 5Ј end-labeled with T4 polynucleotide kinase (PNK) (Promega) and [␥- 32 P]ATP (6,000 Ci/ mmol) (Perkin Elmer, Boston, Mass.). The primers were labeled in a 40-l reaction mixture containing 4 l of 10ϫ PNK buffer, 2 l of primer (10 pmol/l), 5 l of [␥-32 P]ATP, 2 l of T4 PNK, and 27 l of DEPC-H 2 O. Primer extension was performed following the protocol of the primer extension system (Promega).…”

“…As a member of the neurotropic alphaherpesvirus subfamily, VZV can establish latency in the dorsal root ganglia following primary infection (4). Latent VZV DNA is predominantly localized in the neurons, although some researchers also identified viral sequences in nonneuronal satellite cells (14,22,23,32,37). Data obtained from human ganglia and animal models indicate that during latent infection, a small subset of lytic viral genes is expressed while most of the VZV genome is transcriptionally quiescent.…”

The Glycoprotein B Cytoplasmic Domain Lysine Cluster Is Critical for Varicella-Zoster Virus Cell-Cell Fusion Regulation and Infection

“…They have been observed in skin lesions and trigeminal ganglia taken at autopsy from individuals afflicted with zoster at time of death (7,8). The fusion of infected satellite cells and neurons observed in the cadaver ganglia and in the human dorsal root ganglia xenograft model has implicated the damage to the fused cells as a potential factor contributing to PHN after VZV reactivation in the sensory nerve ganglia (9,10). Understanding and defining the mechanisms that govern VZV syncytium formation could lead to the development of effective PHN treatments.…”