Recombinant Varicella-Zoster Virus Glycoproteins E and I: Immunologic Responses and Clearance of Virus in a Guinea Pig Model of Chronic Uveitis

Kimura, Hiroshi; Wang, Yun; Pesnicak, Lesley; Cohen, Jeffrey I.; Hooks, John J.; Straus, Stephen E.; Williams, Russell B.

doi:10.1086/515638

Cited by 40 publications

(32 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…4,5 We have established a system to quantify CMV, EBV, and varicella-zoster virus (VZV) genomes using a 'real-time' quantitative PCR method. [6][7][8] This method measures the accumulation of PCR products with a fluorogenic probe and real-time laser scanning in a 96-well plate, and has a very wide dynamic range. 9 Since this assay does not require additional handling, such as electrophoresis, faster assays with a higher throughput are possible.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Monitoring four herpesviruses in unrelated cord blood transplantation

Tanaka

Kimura

Hoshino

et al. 2000

Bone Marrow Transplant

View full text Add to dashboard Cite

Summary:Cord blood transplantation, which has lower risk of graft-versus-host disease than bone marrow transplantation, might have higher risk of infections. A system to quantify four herpesviruses, CMV, human herpesvirus 6 (HHV6), EBV, varicella-zoster virus using the realtime PCR assay was established and applied for prospective viral load monitoring in three recipients undergoing cord blood transplantation. CMV and HHV6 were detected in peripheral blood from all three recipients, while EBV was detected in two. Varicellazoster virus was not detected at all. At the peak of HHV6 or CMV, each patient showed virus-related symptoms. During the pre-transplant period, CMV DNA was detected in two recipients who later developed CMV-related diseases. These observations indicate that our system is not only useful for managing herpesviruses infections in transplant recipients, but also a powerful method for clarifying the relationships between the viral load and clinical symptoms. Bone Marrow Transplantation (2000) 26, 1193-1197. Keywords: real-time PCR; CMV; human herpesvirus 6 (HHV6); EBV; cord blood transplantation (CBT) Recently, the number of cord blood transplants (CBT) from related and unrelated donors has increased to treat malignant and nonmalignant disorders. The hematopoietic stem cells found in cord blood possess higher proliferative potential than adult bone marrow cells. 1 CBT also has a lower risk for severe GVHD than bone marrow or peripheral blood stem cell transplantation. 1 On the other hand, recipients of CBT may have higher risk of infections than recipients of other hematopoietic stem cell transplants (HSCT) because the cord blood contains few memory T cells to respond to exogenous antigens. Nuckols 2 reported that infectious complications were frequently encountered at autopsy of CBT recipients. It was also reported that a positive serostatus for CMV was associated with worsened survival in CBT recipients. 3 Meanwhile, herpesvirus infections cause serious compli- cations in the management of HSCT recipients. Herpesviruses latently infect many individuals and become reactivated when the immune system is suppressed. Therefore, quantitative analysis is essential to diagnose symptomatic herpesvirus infections. 4,5 We have established a system to quantify CMV, EBV, and varicella-zoster virus (VZV) genomes using a 'real-time' quantitative PCR method. [6][7][8] This method measures the accumulation of PCR products with a fluorogenic probe and real-time laser scanning in a 96-well plate, and has a very wide dynamic range. 9 Since this assay does not require additional handling, such as electrophoresis, faster assays with a higher throughput are possible. In this study, we newly established a system for human herpesvirus 6 (HHV6) and quantified four herpesviruses simultaneously in three recipients who underwent unrelated CBT. With this method, we observed how each viral burden correlated with its clinical manifestations in these recipients. Patients and methods Patient characteristicsThree recipients of ...

show abstract

Section: Discussionmentioning

confidence: 99%

“…[6][7][8] The PCR primers and probe for HHV6 were selected from the U31 gene (large tegment protein). The upstream primer was 5′-TTTGCAGTCATCACGAT-CGG-3′, and downstream primer was 5′-AGAGCGACAAATTGGAGGTTTC-3′.…”

Section: Real-time Quantitative Pcr Assaymentioning

confidence: 99%

Monitoring four herpesviruses in unrelated cord blood transplantation

Tanaka

Kimura

Hoshino

et al. 2000

Bone Marrow Transplant

View full text Add to dashboard Cite

show abstract

“…Studies with gE subunit vaccines already demonstrated the induction of specific B and T-cell responses in animals. 42,43 Interestingly, preliminary results from an open randomized study in adults exploring a recombinant gE vaccine with adjuvant for zoster prevention demonstrated more robust cellular (i.e. CD4) and humoral immune responses in older adults compared to the application of a live varicella vaccine.…”

Section: Discussionmentioning

confidence: 99%

Varicella-zoster virus glycoproteins B and E are major targets of CD4+ and CD8+ T cells reconstituting during zoster after allogeneic transplantation

Kleemann

Distler²,

Wagner³

et al. 2011

Haematologica

View full text Add to dashboard Cite

The online version of this article has a Supplementary Appendix. BackgroundAfter allogeneic hematopoietic stem-cell transplantation patients are at increased risk for herpes zoster as long as varicella-zoster virus specific T-cell reconstitution is impaired. This study aimed to identify immunodominant varicella-zoster virus antigens that drive recovery of virusspecific T cells after transplantation. Design and MethodsAntigens were purified from a varicella-zoster virus infected cell lysate by high-performance liquid chromatography and were identified by quantitative mass spectrometric analysis. To approximate in vivo immunogenicity for memory T cells, antigen preparations were consistently screened with ex vivo PBMC of varicella-zoster virus immune healthy individuals in sensitive interferon-γ ELISpot assays. Candidate virus antigens identified by the approach were genetically expressed in PBMC using electroporation of in vitro transcribed RNA encoding full-length proteins and were then analyzed for recognition by CD4 + and CD8 + memory T cells. ResultsVaricella-zoster virus encoded glycoproteins B and E, and immediate early protein 62 were identified in immunoreactive lysate material. Predominant CD4 + T-cell reactivity to these proteins was observed in healthy virus carriers. Furthermore, longitudinal screening in allogeneic stem-cell transplantation patients showed strong expansions of memory T cells recognizing glycoproteins B and E after onset of herpes zoster, while immediate early protein 62 reactivity remained moderate. Reactivity to viral glycoproteins boosted by acute zoster was mediated by both CD4 + and CD8 + T cells. ConclusionsOur data demonstrate that glycoproteins B and E are major targets of varicella-zoster virus specific CD4 + and CD8 + T-cell reconstitution occurring during herpes zoster after allogeneic stemcell transplantation. Varicella-zoster virus glycoproteins B and E might form the basis for novel non-hazardous zoster subunit vaccines suitable for immunocompromised transplant patients.

show abstract

“…Recently, we established a quantitative real-time polymerase chain reaction (PCR) assay to quantify the VZV genome copies (8). Using this assay, we observed that there was a higher occurrence of viremic VZV in varicella than in zoster, and that acyclovir treatment marked-*Address correspondence to Dr. Yoshinori Ito, Department of PediatricslDevelopmental Pediatrics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 466-8550, Japan.…”

mentioning

confidence: 99%

“…Whole blood was obtained from the patients, and plasma and PBMC were separated by density gradient with Ficoll-Paque (Pharmacia Biotech, Uppsala, Sweden). The real-time quantitative PCR assay was performed using a TaqMan PCR kit and a Model 7700 Sequence Detector (PE Applied Biosystems, Foster City, U.S.A.), as previously described (8,9). Primers to amplify the glycoprotein B (gB) and ORF62 genes were selected.…”

mentioning

confidence: 99%

Investigation of Varicella‐Zoster Virus DNA in Lymphocyte Subpopulations by Quantitative PCR Assay

Ito

Kimura

Hara

et al. 2001

Microbiology and Immunology

View full text Add to dashboard Cite

Abstract:To investigate the nature of viremia during the acute phase of varicella, we studied the viral load in nine otherwise healthy children with varicella. Plasma and peripheral blood mononuclear cells (PBMC) were obtained, then PBMC were divided into CD4+T, CD8+T, and B lymphocytes and monocyWmacrophage fractions. The viral DNA in each component was quantified using a real-time quantitative polymerase chain reaction assay. Varicella-zoster virus (VZV) DNA was detected in plasma, PBMC and all subpopulations. The amount of viral DNA was similar in each PBMC subpopulation, suggesting that each lymphocyte fraction and monocytes carry similar amounts of VZV DNA during viremia.Key words: Varicella-zoster virus, Lymphocyte subpopulations, Quantitative PCR assay Varicella (chickenpox) is characterized by febrile illness with pruritic vesicles, and is the result of primary infection with varicella-zoster virus (VZV). The pathogenesis is as follows: virus entry at the respiratory mucosa and primary viremia are believed to occur; then secondary viremia of greater magnitude occurs; finally, the rash results. Understanding the nature of the viremia is thought to be important in order to clarify the pathophysiology of varicella infections and complications of the disease (I, 7). Peripheral blood mononuclear cell (PBMC)-associated viremia has been demonstrated just before and after the onset of disease (2,5,6,13,15,16). Determining the phenotypes of the PBMC subpopulations is complicated by the low frequency of positive cells and the difficulty in achieving highly purified preparations of subpopulations.Recently, we established a quantitative real-time polymerase chain reaction (PCR) assay to quantify the VZV genome copies (8). Using this assay, we observed that there was a higher occurrence of viremic VZV in varicella than in zoster, and that acyclovir treatment marked-*Address correspondence to Dr. Yoshinori Ito, Department of PediatricslDevelopmental Pediatrics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 466-8550, Japan. Fax: 81-52-744-2974. E-mail: yyito @med.nagoya-u.ac.jp 267 ly suppressed viremia in varicella (9). In this study, we quantified the viral load in the peripheral blood leukocyte subpopulations to determine which subpopulations harbor VZV during the acute phase of varicella. We also quantified the viral load in plasma to investigate whether viral DNA in blood is restricted to PBMC fraction.Nine healthy children with clinically diagnosed varicella (0 to 3 years old, median: I year) were enrolled in this study. None of these patients had a previous history of varicella. At the time of entry, none of the patients had received oral acyclovir therapy.All the specimens were obtained after informed consent. Blood samples were taken when the patients had clinical indications in the acute phase. Sampling occurred 2 to 5 days after the onset (mean: 2.3 days). Whole blood was obtained from the patients, and plasma and PBMC were separated by density gradient with Ficoll-P...

show abstract

Recombinant Varicella-Zoster Virus Glycoproteins E and I: Immunologic Responses and Clearance of Virus in a Guinea Pig Model of Chronic Uveitis

Cited by 40 publications

References 32 publications

Monitoring four herpesviruses in unrelated cord blood transplantation

Monitoring four herpesviruses in unrelated cord blood transplantation

Varicella-zoster virus glycoproteins B and E are major targets of CD4+ and CD8+ T cells reconstituting during zoster after allogeneic transplantation

Investigation of Varicella‐Zoster Virus DNA in Lymphocyte Subpopulations by Quantitative PCR Assay

Contact Info

Product

Resources

About