Paula W. Annunziato

Persistence of Immunity to Live Attenuated Varicella Vaccine in Healthy Adults

The varicella vaccine was approved in 1995 for use in healthy varicella-susceptible children and adults. Long-term immunity in 461 healthy adults who were enrolled in varicella vaccine trials in 1979-1999 were studied. Forty vaccinees (9%), including 19 (21%) of 89 vaccinees with household exposure (HHE) to chickenpox, developed breakthrough chickenpox 8 weeks to 11.8 years (mean, 3.3 years) after vaccination. The median number of skin lesions among the 36 untreated vaccinees was 20 (range, 1-240 lesions), and the number of lesions was essentially the same with time since vaccination. Breakthrough chickenpox was mild, even among vaccinees who did not have seroconversion or those recipients who lost detectable antibody. Lower varicella-zoster virus (VZV) antibody titers measured within 3 months of vaccination as well as at the time of HHE were associated with an increased risk of breakthrough disease. This study demonstrated that the varicella vaccine was effective in providing adults with long-term protection from serious VZV disease.

Open Reading Frame S/L of Varicella-Zoster Virus Encodes a Cytoplasmic Protein Expressed in Infected Cells

ABSTRACT We report the discovery of a novel gene in the varicella-zoster virus (VZV) genome, designated open reading frame (ORF) S/L. This gene, located at the left end of the prototype VZV genome isomer, expresses a polyadenylated mRNA containing a splice within the 3′ untranslated region in virus-infected cells. Sequence analysis reveals significant differences between the ORF S/Ls of wild-type and attenuated strains of VZV. Antisera raised to a bacterially expressed portion of ORF S/L reacted specifically with a 21-kDa protein synthesized in cells infected with a VZV clinical isolate and with the original vaccine strain of VZV (Oka-ATCC). Cells infected with other VZV strains, including a wild-type strain that has been extensively passaged in tissue culture and commercially produced vaccine strains of Oka, synthesize a family of proteins ranging in size from 21 to 30 kDa that react with the anti-ORF S/L antiserum. MeWO cells infected with recombinant VZV harboring mutations in the C-terminal region of the ORF S/L gene lost adherence to the stratum and adjacent cells, resulting in an altered plaque morphology. Immunohistochemical analysis of VZV-infected cells demonstrated that ORF S/L protein localizes to the cytoplasm. ORF S/L protein was present in skin lesions of individuals with primary or reactivated infection and in the neurons of a dorsal root ganglion during virus reactivation.

Evidence of Latent Varicella-Zoster Virus in Rat Dorsal Root Ganglia

Latent varicella-zoster virus (VZV) was studied in ganglia of rats that had been inoculated subcutaneously with either a high-passaged wild-type, a low-passaged wild-type, or the vaccine strain of virus using in situ hybridization. Nine of 11 rats injected with virus and no control rats developed serum VZV antibodies as demonstrated by fluorescent antibody membrane antigen. Polymerase chain reaction 2 weeks following inoculation did not detect viremia in the rats. VZV was detected by in situ hybridization in ganglia of 10 of the 11 infected rats but not in ganglia of the control rats. The distribution of VZV DNA is identical to that seen in humans; satellite cells and neurons contain VZV DNA. Although all animals received unilateral injections of virus, VZV DNA was in ipsilateral and contralateral ganglia in 6 animals, suggesting that virus replication and viremia had occurred.

Congenital Varicella-Zoster Virus Infection and Barrett's Esophagus

Congenital varicella syndrome is a rare complication of varicella-zoster virus (VZV) infection during pregnancy. An infant was exposed to VZV at 18.5 weeks of gestation and had eye and skin abnormalities at birth and persistent feeding difficulties, prompting esophageal biopsies at 12 days and 20 and 20.5 months of age. Esophageal tissues demonstrated specialized intestinal metaplasia (Barretts esophagus). VZV DNA (in situ hybridization) and proteins (immunohistochemistry and polymerase chain reaction) were found in esophageal epithelial cells adjacent to the Barretts lesion. Immediate-early 63 protein (IE63) of VZV was demonstrated in the day 12 specimen, and IE62 and the late VZV glycoprotein E (gE) were found in the 20-month specimen. Clinical and endoscopic improvement followed fundoplication and acyclovir therapy, but VZV DNA and IE62 persisted in esophageal tissue. These findings associate VZV with specialized intestinal metaplasia within the esophagus and suggest a novel site for either latent or active VZV infection.

In situ hybridization detection of varicella zoster virus in paraffin-embedded skin biopsy samples

BACKGROUNDnWhen virologic and molecular diagnostic techniques are unavailable, the diagnosis of varicella zoster virus (VZV) infection depends on clinical criteria and histologic evaluation of skin biopsy specimens or Tzank preparations. These methods can misdiagnose chickenpox and zoster, particularly when the clinical manifestations are atypical.nnnOBJECTIVEnTo improve diagnosis in these settings, we developed an in situ hybridization technique for the detection of VZV utilizing a fluorescein-labeled oligonucleotide probe visualized with anti-fluorescein alkaline phosphatase-conjugated antibody.nnnSTUDY DESIGNnWe retrospectively examined 26 paraffin-embedded skin biopsy specimens with histologic features consistent with VZV or herpes simplex virus (HSV) infection and 11 control cases by in situ hybridization. In situ hybridization for VZV and HSV-1 was compared with polymerase chain reaction (PCR) for VZV and HSV-1 and clinical and histologic examination.nnnRESULTSnThirteen of the 26 study cases and two of the 11 control cases were positive for VZV by in situ hybridization. When compared with PCR, in situ hybridization was 92% sensitive and 88% specific. When compared with clinical diagnosis, in situ hybridization was 86% sensitive and 87% specific. All cases of chickenpox had VZV-positive inflammatory cells in the dermis but this finding was less frequent among the cases of zoster.nnnCONCLUSIONSnThis in situ hybridization technique is a sensitive and specific method for the diagnosis of VZV in skin lesions that is applicable to most histopathology laboratory settings. In addition, in situ hybridization reveals individual infected cells and may provide insight into the pathogenesis of VZV skin infection.

Varicella-zoster virus proteins in skin lesions: implications for a novel role of ORF29p in chickenpox.

ABSTRACT Skin biopsy samples from varicella-zoster virus (VZV)-infected patients examined by immunohistochemistry demonstrated VZV replication in nonepithelial cell types. ORF29p, a nonstructural nuclear protein, was found in nerves of two of six patients with chickenpox. In tissue culture, ORF29p was secreted by VZV-infected fibroblasts. Extracellular ORF29p can be taken up through endocytosis by human neurons, implying a novel role for this protein in pathogenesis.

New Developments in Pertussis Vaccines

The incidence of pertussis disease declined following widespread immunisation earlier in the twentieth century. However, adverse affects associated with whole-cell pertussis vaccines and recent concern about their efficacy has lead to poor acceptance in a number of countries. Administration of whole-cell pertussis vaccine is associated with a number of undesirable effects such as fever, local reactions and prolonged crying episodes, in addition to rare but serious acute neurological sequelae that may occur in 1 in 100 000 to 1 000 000 vaccinations. Furthermore, recent increases in the incidence of pertussis, even among vaccinated individuals, raise questions regarding the efficacy of the whole-cell vaccine. A number of acellular pertussis vaccines have been developed to improve vaccine efficacy and decrease vaccine-associated adverse effects. Acellular pertussis vaccines consist of purified, inactivated pertussis antigens. Two types of acellular vaccines are widely used in Japan and are gaining acceptance in other countries. The B-type vaccine contains equal amounts of pertussis toxin and filamentous haemagglutinin. The T-type vaccine contains larger amounts of filamentous haemagglutinin and smaller amounts of pertussis toxin, pertactin and other agglutinogens. In studies conducted in Japan, the US and Sweden, these vaccines have been associated with few adverse effects and elicit a measurable antibody titre to their components. Efficacy studies evaluating these vaccines have also been promising, although no study has compared the efficacy of whole-cell vaccine with the efficacy of acellular pertussis vaccine, and the quantities of acellular vaccine antigens required for protection have not been determined. Studies to answer these questions are ongoing.

Varicella zoster virus in human and rat tissue specimens

The limited supple of appropriate tissues for study has been an impediment to investigations of varicella zoster virus (VZV) latency. Human dorsal root ganglia (DRG) harboring latent virus are not plentiful and are not amenable to manipulation for studying the events surrounding the establishment, maintenance, and cessation of latency. An alternative to studies in human DRG is the rat model of latency, which appears to provide a reliable method of investigating VZV latency. Other alternatives include studies in other human tissues involved in VZV pathogenesis. In order to improve our understanding of the establishment and cessation of latency, we performed comparative immunohistochemical analysis of chickenpox and zoster skin lesions. This analysis revealed that during primary infection and reactivation productive VZV infection occurs in a variety of cell types and that the major VZV DNA binding protein, ORF29p, is present in peripheral axons early during the course of chickenpox. VZV latency was studied in the rat model by in situ hybridization and compared with similar studies performed in human DRG containing latent virus, confirming that VZV DNA persists in the same sites in DRG of the two species.