Varicella-zoster Virus Vaccine Strain Research Articles

The impact of varicella vaccination in the United States

The addition of varicella vaccine to the universal childhood immunization schedule in the United States in 1995 can be seen as a bold step. Shown to be safe and efficacious against varicella in extensive prelicensure studies, it is nonetheless the first vaccine against a herpesvirus and, furthermore, it is a live, attenuated vaccine. Both wild-type and vaccine strain varicella zoster virus (VZV) are noteworthy for their ability to establish latent infection within the host, with the subsequent possibility of reactivation. Therefore, at the population level, a successful vaccination program could result in the eventual displacement of wild-type VZV by the attenuated vaccine virus. The immediate objective of universal vaccination, however, was to reduce the significant morbidity and mortality associated with primary VZV infection. Data now accumulating suggest that the varicella vaccine as used in the United States has so far been highly effective. The challenge for the future is to predict how the resulting substantial reduction in circulation of VZV will affect immunity among both vaccinees and the unvaccinated. Vaccination strategies likely will need to be adjusted as the epidemiology of VZV in the United States continues to evolve.

Varicella-zoster virus infection of human neural cells in vivo.

Varicella-zoster virus (VZV) establishes latency in sensory ganglia and causes herpes zoster upon reactivation. These investigations in a nonobese diabetic severe combined immunodeficient mouse-human neural cell model showed that VZV infected both neurons and glial cells and spread efficiently from cell to cell in vivo. Neural cell morphology and protein synthesis were preserved, in contrast to destruction of epithelial cells by VZV. Expression of VZV genes in neural cells was characterized by nuclear retention of the major viral transactivating protein and a block in synthesis of the predominant envelope glycoprotein. The attenuated VZV vaccine strain retained infectivity for neurons and glial cells in vivo. VZV gene expression in differentiated human neural cells in vivo differs from neural infection by herpes simplex virus, which is characterized by latency-associated transcripts, and from lytic VZV replication in skin. The chimeric nonobese diabetic severe combined immunodeficient mouse model may be useful for investigating other neurotropic human viruses.

A real-time PCR assay for the detection of varicella-zoster virus DNA and differentiation of vaccine, wild-type and control strains

Varicella-zoster vaccine is a live attenuated virus. It is, therefore, necessary to have a test to differentiate vaccine from wild-type varicella-zoster virus (VZV) strains for the investigation of varicella or zoster-like rash illness in individuals vaccinated previously. In addition, it is necessary to have a rapid VZV assay for use in the context of smallpox bioterrorism laboratory testing. Using specific primers and hybridization probes, a rapid method to differentiate vaccine strain VZV from wild-type VZV was developed based on the presence or absence of a Pst I restriction site within open reading frame (ORF) 38. Using this ORF 38 assay in conjunction with a similar previously described ORF 62 assay allows for further differentiation of vaccine strain, wild-type and a laboratory control strain (Ellen) VZV. This is accomplished because Ellen VZV is similar to wild-type VZV with respect to the ORF 38 assay but is similar to vaccine strain VZV with respect to the ORF 62 assay. The hybridization probes for each ORF are labeled with different fluorescent tags thus allowing both assays to be run simultaneously in a single tube. Both assays demonstrate a high degree of specificity for VZV and can reliably detect between 10 and 100 copies of VZV DNA. Thus, the real-time polymerase chain reaction (PCR) assay for VZV described below provides a rapid assay allowing the simultaneous differentiation of vaccine, wild-type and laboratory control strains of VZV.

Use of a rodent model to show that varicella-zoster virus ORF61 is dispensable for establishment of latency.

Varicella-zoster virus (VZV) results in a latent infection in humans after primary infection. Latency has also been established in guinea pigs and rats after inoculation with the virus. It was found that infection of cotton rats with the Oka vaccine strain of VZV results in a latent infection. To begin to identify which genes are required for latency, we infected cotton rats with VZV strain Oka that is deleted for ORF61. ORF61 protein transactivates certain VZV promoters and enhances the infectivity of viral DNA in transient transfections. Deletion of ORF61 results in abnormal syncytia and impairs the growth of VZV in vitro. Inoculation of cotton rats with ORF61-deleted Oka virus resulted in latent VZV infection in the nervous system similar to that seen for animals infected with parental virus. Thus, the cotton rat can be used to study the ability of mutants in the Oka vaccine strain of VZV to establish latent infection.

The out of Africa model of varicella-zoster virus evolution: single nucleotide polymorphisms and private alleles distinguish Asian clades from European/North American clades

Until 1998, varicella-zoster virus (VZV) was generally considered sufficiently stable to allow the use of a single sequenced virus (VZV-Dumas) as a consensual representation of the world VZV genotype. But recent investigations have uncovered a gE mutant virus called VZV-MSP with a second genotype and a distinguishable accelerated cell spread phenotype. A subsequent study suggested that single nucleotide polymorphisms (SNPs) could be applied toward the genetic analysis of the VZV genome. To further assess the scope of genetic variation in the VZV genome on a worldwide basis, we carried out an extensive SNP analysis of structural glycoprotein genes gB, gE, gH, gI, gL, as well as the IE62 regulatory gene in viruses collected from Western Europe, North America and Asia, including the VZV vaccine strain. The SNP data showed segregation of viral isolates of Asian origin from those of Western ancestry into distinct phylogenetic clades. Unexpectedly, however, VZV from Thailand segregated with VZV from Iceland and the United States, i.e. it was more Western than Asian in nature. Further, SNP analysis disclosed strikingly unusual genotypes, e.g. gH genes with up to five missense mutations and gL genes with insertions of an in-frame methionine codon. In summary, these VZV genomic analyses have shown that individual VZV strains, like closely related human beings, have distinctive SNP profiles containing private alleles within just five VZV genes ( gB, gH, gE, gL and IE62) that provide a fingerprint to localize ancestry of the viral strain.

Open reading frame S/L of varicella-zoster virus encodes a cytoplasmic protein expressed in infected cells.

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.

Viral strain identification in varicella vaccinees with disseminated rashes.

Approximately 15% of recipients of live attenuated varicella vaccine may develop mild breakthrough varicella months to years after immunization. Although some vaccinees will develop zoster, it is less common in recipients of vaccine than in those who have had natural varicella. To determine the varicella-zoster virus (VZV) strain responsible for breakthrough varicella and zoster in recipients of varicella vaccine. A PCR assay capable of distinguishing wild-type from vaccine strain VZV was performed on samples from skin lesions from vaccinees with breakthrough varicella and zoster. All of 57 vaccinees with breakthrough varicella, clinically diagnosed on the basis of a generalized maculopapular or vesicular rash, in which there was amplifiable DNA [corrected], had wild-type VZV infection based on analysis of viral DNA. The Oka vaccine strain of VZV was not identified in any of these cases. In contrast, in 32 patients with zosteriform rashes, the vaccine strain was identified in 22 samples, and the wild-type strain was identified in 10 samples. Wild-type virus was identified in all generalized rashes occurring after the immediate 6-week postvaccination period. When reactivation of vaccine strain occurred, it presented as typical zoster. We find no evidence that reactivation of vaccine virus occurs with the clinical picture of generalized rash.

Postlicensure safety surveillance for varicella vaccine.

Since its licensure in 1995, the extensive use of varicella vaccine and close surveillance of the associated anecdotal reports of suspected adverse effects provide the opportunity to detect potential risks not observed before licensure because of the relatively small sample size and other limitations of clinical trials. To detect potential hazards, including rare events, associated with varicella vaccine, and to assess case reports for clinical and epidemiological implications. Postlicensure case-series study of suspected vaccine adverse events reported to the US Vaccine Adverse Event Reporting System (VAERS) from March 17, 1995, through July 25, 1998. Numbers of reported adverse events, proportions, and reporting rates (reports per 100,000 doses distributed). VAERS received 6574 case reports of adverse events in recipients of varicella vaccine, a rate of 67.5 reports per 100,000 doses sold. Approximately 4% of reports described serious adverse events, including 14 deaths. The most frequently reported adverse events were rashes, possible vaccine failures, and injection site reactions. Misinterpretation of varicella serology after vaccination appeared to account for 17% of reports of possible vaccine failures. Among 251 patients with herpes zoster, 14 had the vaccine strain of varicella zoster virus (VZV), while 12 had the wild-type virus. None of 30 anaphylaxis cases was fatal. An immunodeficient patient with pneumonia had the vaccine strain of VZV in a lung biopsy. Pregnant women occasionally received varicella vaccine through confusion with varicella zoster immunoglobulin. Although the role of varicella vaccine remained unproven in most serious adverse event reports, there were a few positive rechallenge reports and consistency of many cases with syndromes recognized as complications of natural varicella. Most of the reported adverse events associated with varicella vaccine are minor, and serious risks appear to be rare. We could not confirm a vaccine etiology for most of the reported serious events; several will require further study to clarify whether varicella vaccine plays a role. Education is needed to ensure appropriate use of varicella serologic assays and to eliminate confusion between varicella vaccine and varicella zoster immunoglobulin. JAMA. 2000;284:1271-1279

Humoral immunoresponse to varicella-zoster virus pernasally coadministered with Escherichia coli enterotoxin in mice

It is evaluated whether Escherichia coli enterotoxin is useful for induction of immunity to varicella-zoster virus (VZV) as a mucosal adjuvant in mice. When a commercially available live varicella vaccine (Oka strain) and toxin were administered simultaneously via a nasal route three times at 2 or 6 month intervals, an antibody neutralizing VZV was detected in half or all of the mice vaccinated, respectively. The antibody specific to the vaccine strain of VZV reacted to five proteins, molecular weights of which were 110 K, 100 K, 62 K, 54 K and 46 K. These proteins were composed of glycosylated products of all kinds of glycoproteins. These results suggest that although a nasal administration of the vaccine without the adjuvant has little immunogenicity in mice, the simultaneous administration of the live vaccine and the toxin over a long period induces a specific humoral immunity to VZV.

Polymerase chain reaction and restriction fragment length polymorphism analysis of varicella-zoster virus isolates from the United States and other parts of the world.

A polymerase chain reaction (PCR) assay that identifies and differentiates wild-type (wt) and vaccine strains of varicella-zoster virus (VZV) was used to determine if VZV strains with restriction fragment length polymorphisms resembling those of the Japanese Oka vaccine strain were present in the wt pool outside of Japan. Virus samples (n = 114) from patients with chickenpox and zoster from various parts of the United States and Australia were analyzed. The assay correctly identified 113 samples as wt strain. The 1 sample identified as Oka vaccine strain came from a child with leukemia who developed a vaccine-associated rash after receiving the live attenuated varicella vaccine. At this point, there is no evidence that wt strains resembling the vaccine are circulating outside of Japan. This indicates that this PCR assay can be utilized to distinguish rashes due to vaccine and wt VZV.

Identification of the Oka strain of the live attenuated varicella vaccine from other clinical isolates by molecular epidemiologic analysis.

A method was developed to distinguish the Oka vaccine strain of varicella-zoster virus (VZV) from other clinical isolates. The molecular characteristics of 52 clinical isolates from varicella or zoster patients with no history of VZV vaccination and the Oka strain, including vaccine and parental viruses, were analyzed by PstI cleavage of the PstI site-less (PSL) region. This was followed by single-strand conformational polymorphism (SSCP) after polymerase chain reaction amplification of repeating region 2 (R2). Most of the clinical isolates tested, especially recent isolates, had a PstI site in the PSL region, but the Oka strain did not. The SSCP patterns of R2 in Oka strain virus differed from those of other viruses. These results suggest that analysis of the PstI site followed by SSCP of R2 will be useful for identifying the Oka vaccine virus in isolates.

Herpes zoster after varicella immunization

cine was licensed in the United States in 1995. The vaccine is safe, immunogenic, and highly protective against severe varicella in healthy children and adults and in certain immunocompromised patients.1-3 It was initially hoped that herpes zoster might not occur after immunization. However, Oka vaccine-type varicella-zoster virus can cause latent infection in children with acute lymphocytic leukemia.4,5 We describe herpes zoster caused by vaccine strain varicella-zoster virus in a child.

Immunization with recombinant varicella-zoster virus expressing herpes simplex virus type 2 glycoprotein D reduces the severity of genital herpes in guinea pigs.

Varicella-zoster virus (VZV) is an attractive candidate for a live-virus vector for the delivery of foreign antigens. The Oka vaccine strain of VZV is safe and effective in humans, and recombinant Oka VZV (ROka) can be generated by transfecting cells with a set of overlapping cosmid DNAs. By this method, the herpes simplex virus type 2 (HSV-2) glycoprotein D (gD2) gene was inserted into an intergenic site in the unique short region of the Oka VZV genome. Expression of gD2 in cells infected with the recombinant Oka strain VZV (ROka-gD2) was confirmed by antibody staining of fixed cells and by immunoblot analysis. Immune electron microscopy demonstrated the presence of gD2 in the envelope of ROka-gD2 virions. The ability of ROka-gD2 to protect guinea pigs against HSV-2 challenge was assessed by inoculating animals with three doses of uninfected human fibroblasts, fibroblasts infected with ROka VZV, or fibroblasts infected with ROka-gD2. Neutralizing antibodies specific for HSV-2 developed in animals immunized with ROka-gD2. Forty days after the third inoculation, animals were challenged intravaginally with HSV-2. Inoculation of guinea pigs with ROka-gD2 significantly reduced the severity of primary HSV-2 infection (P < 0.001). These experiments demonstrate that the Oka strain of VZV can be used as a live virus vector to protect animals from disease with a heterologous virus.

Infection of human fetal dorsal root neurons with wild type varicella virus and the Oka strain varicella vaccine

The relative ability of a varicella-zoster virus (VZV) clinical isolate and a live attenuated VZV vaccine strain (Oka) to infect human neurons was determined in vitro. VZV infection of neurons prepared in culture from dorsal root ganglia of fetuses was assessed using an infectious center assay. Cultures were infected with 50-5,000 pfu of either VZV and assayed at either 24 or 48 hours post-VZV infection. Cultures infected with the clinical VZV isolate had seven-fold more infected neurons than cultures infected with the vaccine strain VZV.

Restriction fragment differences between the genomes of the Oka varicella vaccine virus and american wild‐type varicella‐zoster virus

The Oka vaccine strains of varicella-zoster virus (VZV) have a significantly different BgII DNA restriction pattern from that of American wild-type isolates of VZV. This difference consists primarily of an additional BgII site, which lies within the BamHI "D" fragment. In conjunction with a study of the efficacy of an experimental Merck/Oka VZV vaccine, the area of the genome from which the most marked restriction pattern alteration arises was studied more closely to determine if there are other significant differences between the Oka strains and American wild-type strains. BamHI "D" fragments from the DNA of the Oka parent strain (the progenitor of the vaccine strain), the RIT/Oka vaccine strain (a derivative of the Oka parent strain), the Merck/Oka vaccine strain, and the EF strain (an American wild type), were submitted to extensive endonuclease digestion studies to ascertain if additional unique restriction sites are present in the Oka parent or vaccine strains. The extra BgII restriction site characteristic of the Merck/Oka vaccine strain is also present in the DNA of the parent virus as well as its derivatives and was therefore not produced by the "attenuation" process. No other novel sites were found in the Oka parent or Oka-derived strains in this section of the genome. The Merck/Oka vaccine strain of VZV, despite its Japanese origin, is therefore quite similar to circulating American varicella-zoster virus strains. Varicella-zoster virus DNA, at least in the area of the BamHI D fragment, also appears to be remarkably stable from strain to strain.

Inoculation of guinea pigs with varicella-zoster virus via the respiratory route.

Guinea pigs were inoculated by the respiratory route with wild-type (Cyr) or vaccine (Oka) strain varicella zoster virus (VZV). Wild-type cell-free virus obtained by sonication produced neutralizing antibody responses in steroid-treated animals when given via the intratracheal route, and induced neutralizing antibody as well as a pneumonitis in normal animals when given via the intrabronchial (i.b.) route. A humoral response also followed i.b. instillation of cell-associated wild-type or vaccine strain VZV. Prior i.b. administration of thioglycollate or exposure to hyperoxia altered the number and function of pulmonary macrophages, respectively, but viral susceptibility of the guinea pigs was not enhanced. Both strains of VZV could be isolated from bronchial washings up to 48 hours after i.b. instillation of cell-associated virus, but neither strain was isolated thereafter from cultures of bronchial washings or explanted lung tissues.

Biologic and biophysical markers of a live varicella vaccine strain (Oka): identification of clinical isolates from vaccine recipients.

Possible biologic and biophysical markers of the Oka vaccine strain of varicella-zoster virus (VZV) were explored. The ratio between the infectivity of the vaccine strain in guinea-pig embryo fibroblasts and that in human embryo fibroblasts was consistently higher than the corresponding ratios for wild-type strains; this result seemed to correlate with the passage history of the vaccine strain in guinea-pig embryo fibroblasts. According to profiles of cleavage of digested DNA by various restriction endonucleases, the mobility of one fragment (K) produced by restriction endonuclease Hpa I was unique to the Oka strain. Nine clinical isolates from vaccine recipients (seven with varicella and two with zoster) were examined by both methods (infectivity and cleavage profiles) to determine whether or not these strains were vaccine derived. The laboratory judgments obtained by the two methods were in agreement and were consistent with clinical conclusions concerning five of the strains. Since the test of relative infectivity is simple and convenient, it is suitable for differentiation of the vaccine strain from other wild-type strains; if the results are not conclusive, profiles of cleavage by Hpa I may be helpful in reaching a definite judgment.

Susceptibility of vaccine strains of varicella-zoster virus to antiviral compounds.

Using a plaque reduction assay, we determined the 50% effective doses of six antiviral compounds against low- and high-passage viruses of the KMcC and Oka strains of varicella-zoster virus vaccine. The potency, as indicated by the ranges of 50% effective doses (micrograms per milliliter) of the antiviral compounds, in decreasing order was as follows: (E)-5-(2-bromovinyl)-2'-deoxyuridine, 0.0007 to 0.0035; 1-(2'-flouro-2-deoxy-beta-D-arabinofuranosyl)-5-iodocytosine, 0.0063 to 0.0091; aphidicolin, 0.092 to 0.180; acyclovir, 0.79 to 1.81; vidarabine, 0.62 to 2.10; and phosphonoformic acid, 8.18 to 16.4. Susceptibility to the various antiviral compounds was independent of passage level or strain. These data, along with the available in vivo data, indicate that varicella-zoster virus vaccine infections requiring antiviral therapy most probably would be treated as effectively as would natural varicella infections.

Common expression of varicella-zoster viral glycoprotein antigens in vitro and in chickenpox and zoster vesicles.

Human cells infected with varicella-zoster virus (VZV) produce at least three major virus-specific, immunogenic glycoproteins: gp118, gp98, and gp62. Since glycoproteins gp98 and gp62 were found to be prominent constituents of the infected cell membrane, a murine monoclonal antibody (clone 3B3) that reacted avidly with this glycoprotein complex was selected as a probe for detection of VZV replication in laboratory and clinical settings. Cultured cells of human, simian, and caviid origin, when infected with wild-type isolates as well as laboratory and vaccine strains of VZV all expressed these viral glycoproteins. The monoclonal antibody immunostained the basal and malpighian epithelial layers of a zoster vesicle biopsy specimen and also reacted with all specimens of vesicular cells obtained from epidemiologically unrelated patients with chickenpox and zoster. Thus, these studies demonstrate that the VZV-specific glycoprotein complex gp98/gp62 is highly conserved, abundantly expressed, and easily detected with a monoclonal antibody probe.

Restriction endonuclease analysis of varicella‐zoster vaccine virus and wild‐type dnas

The DNA from several clinical isolates of varicella-zoster virus (VZV) were compared with the DNA from the vaccine strain VZV using three restriction endonucleases: BamHI, BgII, and HpaI. When electrophoresed through an agarose gel, the vaccine DNA digestion pattern was significantly different from the digestion patterns of the wild-type DNAs. Variations in the digestion pattern of the separate clinical isolates were also observed.