A truncated hepatitis E virus ORF2 protein expressed in tobacco plastids is immunogenic in mice

Zhou, Yuan; Lee, Maggie Yuk-Ting; Ng, James Ming-Him; Chye, Mee‐Len; Yip, Wing Ho; Zee, S. Y.; Lam, Eric

doi:10.3748/wjg.v12.i2.306

Cited by 45 publications

(29 citation statements)

References 20 publications

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“…SR1 leaves. The effectiveness of the rice promoter and terminator in tobacco plastids had been demonstrated previously (Leelavathi et al, 2003;Reddy et al, 2002;Zhou et al, 2006). From 20 bombardments, 6 of 10 spectinomycinresistant shoots obtained were confirmed transplastomic by PCR using primer pair Hel-296/Hel-297 (Table I) which amplifies DNA fragments resulting from homologous recombination at the rbcL arm.…”

Section: Estimating the Transplastomic Status Of Transformed Linesmentioning

confidence: 99%

Expression of viral capsid protein antigen against Epstein‐Barr virus in plastids of Nicotiana tabacum cv. SR1

Lee

Zhou

Lung

et al. 2006

Biotech & Bioengineering

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Epstein-Barr virus (EBV) infects nearly 90% of adults worldwide and is the pathogenic source of a broad spectrum of malignancies originating from lymphoid and epithelial cells. Currently, no vaccine has been developed to immunologically inactivate this virus. In infected patients, anti-EBV viral capsid antigen (VCA) immunoglobins represent some of the useful diagnostic markers for carcinoma development. To demonstrate that the EBV VCA antigen can be produced in plants, the plastid genome of tobacco (Nicotiana tabacum cv. SR1) was transformed with a VCA-expressing cassette. The EBV VCA mRNA was actively transcribed in the transplastomic plants and antigen production was detected. This study indicates that plastid transformation could be a promising strategy in EBV VCA antigen production.

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Section: Estimating the Transplastomic Status Of Transformed Linesmentioning

confidence: 99%

Expression of viral capsid protein antigen against Epstein‐Barr virus in plastids of Nicotiana tabacum cv. SR1

Lee

Zhou

Lung

et al. 2006

Biotech & Bioengineering

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“…To prevent HEV infection, an HEV recombinant protein vaccine has been developed [68]. In one of the immunization projects, a large-scale placebo-controlled trial of a vaccine was demonstrated to be 96% effective in the high-risk areas [69]; however, the vaccine was unavailable to commercial use. Consequently, even if the efficacy of HEV vaccine has been demonstrated, its availability still remains a major problem.…”

Section: Prevention and Treatment Of Hepatitis E In Hd Patients And Rmentioning

confidence: 99%

Hepatitis E Virus Infection: A General Review with a Focus on Hemodialysis and Kidney Transplant Patients

Hosseini-Moghaddam¹,

Abbaszadeh²,

Alavian³

et al. 2010

Am J Nephrol

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Background: Hepatitis E virus (HEV) infection is a self-limited viral disease that causes acute hepatitis epidemics in developing countries. The common route of transmission for HEV is supposedly fecal-oral. Serological evidence may be unexpectedly found in hemodialysis (HD) patients and kidney transplant recipients. Although the route of HEV transmission is not usually determined in HD subjects, this virus seems to be transmitted either directly through HD or nosocomially. In this study, we gathered the published information on HEV infection in HD patients and kidney transplant recipients. Methods: For this review, we collected the relevant articles by searching through Medline and Google Scholar from January 1980 up to September 2009. Results: Some variables including older age, low education, living in rural versus urban areas and the duration of HD seem to be risk factors for HEV infection in HD patients. Compared with non-HD subjects, HEV infection may be specifically associated with poor outcome in HD patients. Specific considerations seem to be required to prevent transmission of HEV to HD patients. Conclusion: More extensive investigations are required to determine the disease burden of HEV infection in HD subjects in countries which experience outbreaks of HEV infection.

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“…Since HEV grows poorly in cell culture, the ORF2 gene or fragments thereof have been cloned and expressed in different expression systems, such as prokaryotes, insect cells, animal cells, and transgenic plants [6]. Recently, different recombinant antigens have been used in the design of assays for diagnosing HEV; these assays have optimized the sensitivity and specificity of the antigens in order to provide the best diagnostic test.…”

Section: Introductionmentioning

confidence: 99%

“…HEV belongs to the Hepeviridae family [4]. Its genome consists of a positive single-stranded RNA containing 3 overlapping open reading frames (ORFs), ORF1, ORF2, and ORF3 [5,6]. According to the phylogenetic analysis, HEV has 5 genotypes with different geographical distributions and hosts [7].…”

Section: Introductionmentioning

confidence: 99%

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Development of Enzyme-Linked Immunosorbent Assays Using 2 Truncated ORF2 Proteins for Detection of IgG Antibodies Against Hepatitis E Virus

2014

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Background: Without appropriate culture systems for hepatitis E virus (HEV), sufficient natural viral proteins are difficult to generate for use in serological tests. Therefore, it is important to produce large amounts of HEV recombinant proteins in an economical way. The present study developed ELISAs using 2 truncated forms of the HEV open reading frame (ORF) 2 protein in order to detect anti-HEV IgG in serum samples.Methods: Two truncated forms of the ORF2 protein were expressed in Escherichia coli and were purified by Ni 2+ -chelate-affinity chromatography (Qiagen, Germany). Two ELISAs were developed using these proteins and were compared with DIA.PRO HEV IgG ELISA kit (DIA.PRO. Italy) in 220 serum samples.Results: High yields of the target proteins were obtained through codon optimization. The concentration and purity of the proteins were improved with Amicon filters (EMD Millipore, USA). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting analysis of the resultant proteins showed a protein band of approximately 60 kDa corresponding to ORF2.1 (amino acids 112-660) and a protein band of approximately 55 kDa corresponding to ORF2.2 (amino acids 112-607). Positive agreement, negative agreement, and concordance of the 2 in-house ELISAs compared with DIA.PRO HEV IgG ELISA kit were 87%, 99.5%, and 98.1%, respectively (kappa = 0.899, P = 0.625). Conclusions:The newly developed ELISAs are useful for detecting anti-HEV IgG in serum samples and are highly concordant with DIA.PRO HEV IgG ELISA kit.

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A truncated hepatitis E virus ORF2 protein expressed in tobacco plastids is immunogenic in mice

Abstract: HEV-E2 fragment can be inserted into the plastid genome and the recombinant pE2 antigen derived is antigenic in mice. Hence, plastids may be a novel source for cost-effective production of HEV vaccines.

Cited by 45 publications

References 20 publications

Expression of viral capsid protein antigen against Epstein‐Barr virus in plastids of Nicotiana tabacum cv. SR1

Expression of viral capsid protein antigen against Epstein‐Barr virus in plastids of Nicotiana tabacum cv. SR1

Hepatitis E Virus Infection: A General Review with a Focus on Hemodialysis and Kidney Transplant Patients

Development of Enzyme-Linked Immunosorbent Assays Using 2 Truncated ORF2 Proteins for Detection of IgG Antibodies Against Hepatitis E Virus

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