scholarly journals Characterization of Three Novel Linear Neutralizing B-Cell Epitopes in the Capsid Protein of Swine Hepatitis E Virus

2018 ◽  
Vol 92 (13) ◽  
pp. e00251-18 ◽  
Author(s):  
Yiyang Chen ◽  
Baoyuan Liu ◽  
Yani Sun ◽  
Huixia Li ◽  
Taofeng Du ◽  
...  
Keyword(s):  
B Cell ◽  
Capsid Protein ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Disease Diagnosis ◽  
Antigenic Structure ◽  
Human Populations ◽  
Genotype 3 ◽  
Genotype 4 ◽  
B Cell Epitopes

ABSTRACTHepatitis E virus (HEV) causes liver disease in humans and is thought to be a zoonotic infection, with domestic animals, including swine and rabbits, being a reservoir. One of the proteins encoded by the virus is the capsid protein. This is likely the major immune-dominant protein and a target for vaccination. Four monoclonal antibodies (MAbs), three novel, 1E4, 2C7, and 2G9, and one previously characterized, 1B5, were evaluated for binding to the capsid protein from genotype 4 swine HEV. The results indicated that625DFCP628,458PSRPF462, and407EPTV410peptides on the capsid protein comprised minimal amino acid sequence motifs recognized by 1E4, 2C7, and 2G9, respectively. The data suggested that 2C7 and 2G9 epitopes were partially exposed on the surface of the capsid protein. Truncated genotype 4 swine HEV capsid protein (sp239, amino acids 368 to 606) can exist in multimeric forms. Preincubation of swine HEV with 2C7, 2G9, or 1B5 before addition to HepG2 cells partially blocked sp239 cell binding and inhibited swine HEV infection. The study indicated that 2C7, 2G9, and 1B5 partially blocked swine HEV infection of rabbits better than 1E4 or normal mouse IgG. The cross-reactivity of antibodies suggested that capsid epitopes recognized by 2C7 and 2G9 are common to HEV strains infecting most host species. Collectively, MAbs 2C7, 2G9, and 1B5 were shown to recognize three novel linear neutralizing B-cell epitopes of genotype 4 HEV capsid protein. These results enhance understanding of HEV capsid protein structure to guide vaccine and antiviral design.IMPORTANCEGenotype 3 and 4 HEVs are zoonotic viruses. Here, genotype 4 HEV was studied due to its prevalence in human populations and pig herds in China. To improve HEV disease diagnosis and prevention, a better understanding of the antigenic structure and neutralizing epitopes of HEV capsid protein are needed. In this study, the locations of three novel linear B-cell recognition epitopes within genotype 4 swine HEV capsid protein were characterized. Moreover, the neutralizing abilities of three MAbs specific for this protein, 2C7, 2G9, and 1B5, were studiedin vitroandin vivo. Collectively, these findings reveal structural details of genotype 4 HEV capsid protein and should facilitate development of applications for the design of vaccines and antiviral drugs for broader prevention, detection, and treatment of HEV infection of diverse human and animal hosts.

scholarly journals Characterization of Two Novel Linear B-Cell Epitopes in the Capsid Protein of Avian Hepatitis E Virus (HEV) That Are Common to Avian, Swine, and Human HEVs

2015 ◽  
Vol 89 (10) ◽  
pp. 5491-5501 ◽  
Author(s):  
Xinjie Wang ◽  
Qin Zhao ◽  
Lu Dang ◽  
Yani Sun ◽  
Jiming Gao ◽  
...  
Keyword(s):  
Amino Acid ◽  
B Cell ◽  
Capsid Protein ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Antigenic Structure ◽  
B Cell Epitopes ◽  
Avian Hepatitis E Virus ◽  
Avian Hev ◽  
Linear B

ABSTRACTAntisera raised against the avian hepatitis E virus (HEV) capsid protein are cross-reactive with human and swine HEV capsid proteins. In this study, two monoclonal antibodies (MAbs) against the avian HEV capsid protein, namely, 3E8 and 1B5, were shown to cross-react with the swine HEV capsid protein. The motifs involved in binding both MAbs were identified and characterized using phage display biopanning, peptide synthesis, and truncated or mutated protein expression, along with indirect enzyme-linked immunosorbent assay (ELISA) and Western blotting. The results showed that the I/VPHD motif is a necessary core sequence and that P and H are two key amino acids for recognition by MAb 3E8. The VKLYM/TS motif is the minimal amino acid sequence necessary for recognition by MAb 1B5. Cross-reactivity between the two epitopes and antibodies against avian, swine, and human HEVs in sera showed that both epitopes are common to avian, swine, and human HEVs. In addition, amino acid sequence alignment of the capsid proteins revealed that the key motifs of both novel epitopes are the same in HEVs from different animal species, predicting that they may be common to HEV isolates from boars, rabbits, rats, ferrets, mongooses, deer, and camels as well. Protein modeling analysis showed that both epitopes are at least partially exposed on the surface of the HEV capsid protein. Protective capacity analysis demonstrated that the two epitopes are nonprotective against avian HEV infection in chickens. Collectively, these studies characterize two novel linear B-cell epitopes common to avian, swine, and human HEVs, which furthers the understanding of HEV capsid protein antigenic structure.IMPORTANCEMore and more evidence indicates that the host range diversity of hepatitis E virus (HEV) is a global public health concern. A better understanding of the antigenic structure of the HEV capsid protein may improve disease diagnosis and prevention. In this study, binding site mapping and localization as well as the antigenic biology of two novel linear B-cell epitopes common to several different species of HEV were characterized. These findings partially reveal the antigenic structure of the HEV capsid protein and provide potential applications for the development of diagnostics and interventions for HEV infection.

scholarly journals Molecular tracing of Japan-indigenous hepatitis E viruses

2006 ◽  
Vol 87 (4) ◽  
pp. 949-954 ◽  
Author(s):  
Yasuhito Tanaka ◽  
Kazuaki Takahashi ◽  
Etsuro Orito ◽  
Yoshiyasu Karino ◽  
Jong-Hon Kang ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Demographic History ◽  
Hepatitis E ◽  
Genotype 3 ◽  
Nucleotide Substitutions ◽  
Genotype 4 ◽  
History Of ◽  
The Uk ◽  
Evolutionary Growth ◽  
Rna Polymerase Gene

The ancestor(s) of apparently Japan-indigenous strains of Hepatitis E virus (HEV) was probably of foreign origin, but it remains unclear when and from where it made inroads. In this study, 24 genotype 3 and 24 genotype 4 HEV strains recovered in Japan each showed a significant cluster, clearly distinct from those of foreign strains, in the phylogenetic tree constructed from an 821 nt RNA polymerase gene fragment. The evolutionary rate, approximately 0·8×10−3 nucleotide substitutions per site per year, enabled tracing of the demographic history of HEV and suggested that the ancestors of Japan-indigenous HEV had made inroads around 1900, when several kinds of Yorkshire pig were imported from the UK to Japan. Interestingly, the evolutionary growth of genotype 3 in Japan has been slow since the 1920s, whereas genotype 4 has spread rapidly since the 1980s. In conclusion, these data suggest that the indigenization and spread of HEV in Japan were associated with the popularization of eating pork.

scholarly journals Hepatitis E virus genotype 3 and capsid protein in the blood and urine of immunocompromised patients

2019 ◽  
Vol 78 (3) ◽  
pp. 232-240 ◽  
Author(s):  
Olivier Marion ◽  
Nicolas Capelli ◽  
Sebastien Lhomme ◽  
Martine Dubois ◽  
Mélanie Pucelle ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Immunocompromised Patients ◽  
Genotype 3 ◽  
Virus Genotype

scholarly journals Mapping of linear B cell epitopes on open reading frames 2- and 3-encoded proteins of hepatitis E virus using synthetic peptides

1993 ◽  
Vol 109 (2-3) ◽  
pp. 251-255 ◽  
Author(s):  
Pierre Coursaget ◽  
Yves Buisson ◽  
Nathalie Depril ◽  
Pierre Cann ◽  
Martine Chabaud ◽  
...  
Keyword(s):  
B Cell ◽  
Synthetic Peptides ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Open Reading Frames ◽  
B Cell Epitopes ◽  
Encoded Proteins ◽  
Reading Frames ◽  
Linear B

scholarly journals Human linear B-cell epitopes encoded by the hepatitis E virus include determinants in the RNA-dependent RNA polymerase.

1992 ◽  
Vol 89 (9) ◽  
pp. 3855-3858 ◽  
Author(s):  
M. Kaur ◽  
K. C. Hyams ◽  
M. A. Purdy ◽  
K. Krawczynski ◽  
W. M. Ching ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
B Cell ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Rna Dependent Rna Polymerase ◽  
B Cell Epitopes ◽  
Linear B

scholarly journals Identification of Immunodominant and Conformational Epitopes in the Capsid Protein of Hepatitis E Virus by Using Monoclonal Antibodies

2000 ◽  
Vol 74 (17) ◽  
pp. 8011-8017 ◽  
Author(s):  
Michaela A. Riddell ◽  
Fan Li ◽  
David A. Anderson
Keyword(s):  
Monoclonal Antibodies ◽  
Capsid Protein ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Antigenic Structure ◽  
Conformational Epitopes ◽  
Baculovirus System ◽  
Cross Inhibition ◽  
Murine Monoclonal Antibodies ◽  
Immunodominant Epitopes

ABSTRACT Antibody to the capsid (PORF2) protein of hepatitis E virus (HEV) is sufficient to confer immunity, but knowledge of B-cell epitopes in the intact capsid is limited. A panel of murine monoclonal antibodies (MAbs) was generated following immunization with recombinant ORF2.1 protein, representing the C-terminal 267 amino acids (aa) of the 660-aa capsid protein. Two MAbs reacted exclusively with the conformational ORF2.1 epitope (F. Li, J. Torresi, S. A. Locarnini, H. Zhuang, W. Zhu, X. Guo, and D. A. Anderson, J. Med. Virol. 52:289–300, 1997), while the remaining five demonstrated reactivity with epitopes in the regions aa 394 to 414, 414 to 434, and 434 to 457. The antigenic structures of both the ORF2.1 protein expressed in Escherichia coli and the virus-like particles (VLPs) expressed using the baculovirus system were examined by competitive enzyme-linked immunosorbent assays (ELISAs) using five of these MAbs and HEV patient sera. Despite the wide separation of epitopes within the primary sequence, all the MAbs demonstrated some degree of cross-inhibition with each other in ORF2.1 and/or VLP ELISAs, suggesting a complex antigenic structure. MAbs specific for the conformational ORF2.1 epitope and a linear epitope within aa 434 to 457 blocked convalescent patient antibody reactivity against VLPs by approximately 60 and 35%, respectively, while MAbs against epitopes within aa 394 to 414 and 414 to 434 were unable to block patient serum reactivity. These results suggest that sequences spanning aa 394 to 457 of the capsid protein participate in the formation of strongly immunodominant epitopes on the surface of HEV particles which may be important in immunity to HEV infection.

Hepatitis E virus genotype 4 in a pig farm, Italy, 2013

2014 ◽  
Vol 143 (3) ◽  
pp. 529-533 ◽  
Author(s):  
I. MONNE ◽  
L. CEGLIE ◽  
G. DI MARTINO ◽  
A. NATALE ◽  
S. ZAMPROGNA ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Genotype 3 ◽  
Virus Genotype ◽  
Genotype 4 ◽  
North East ◽  
The North ◽  
Pig Farm ◽  
Swine Population ◽  
First Time

SUMMARYZoonotic strains of hepatitis E virus (HEV) in Europe have been reported to belong to genotypes 3 and 4. In 2012 and 2013, 57 pig farms in Northern Italy that had previously resulted seropositive for HEV were surveyed for the presence of the virus, with positive samples subsequently genotyped. Hepatitis E RNA was identified in 17/57 (29·8%) seropositive farms. Phylogenetic analysis demonstrated that distinct subtypes of genotype 3 were circulating in the north-east of Italy; as well, for the first time in the Italian swine population, genotype 4 was identified and attributed to subtype d.

scholarly journals Detection of Serum Antibodies to Hepatitis E Virus Based on HEV Genotype 3 ORF2 Capsid Protein Expressed in Nicotiana benthamiana

2017 ◽  
Vol 37 (4) ◽  
pp. 313-319 ◽  
Author(s):  
Milena Mazalovska ◽  
Nikola Varadinov ◽  
Tsvetoslav Koynarski ◽  
Ivan Minkov ◽  
Pavel Teoharov ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Nicotiana Benthamiana ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Genotype 3 ◽  
Serum Antibodies

scholarly journals Analyses of Clinical and Biological Data for French and Belgian Immunocompetent Patients Infected With Hepatitis E Virus Genotypes 4 and 3

2021 ◽  
Vol 12 ◽  
Author(s):  
Florence Micas ◽  
Vanessa Suin ◽  
Jean-Marie Péron ◽  
Caroline Scholtes ◽  
Edouard Tuaillon ◽  
...  
Keyword(s):  
Confidence Interval ◽  
Odds Ratio ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Biological Data ◽  
Bivariate Analysis ◽  
Genotype 3 ◽  
Genotype 4 ◽  
Dark Urine ◽  
Immunocompetent Patients

Hepatitis E virus (HEV) genotypes 3 and 4 are the major causes of acute hepatitis in industrialized countries. Genotype 3 is mainly found in Europe and America, while genotype 4 is predominant in Asia. Several Japanese studies have suggested that genotype 4 is more virulent than genotype 3. We investigated this aspect by analyzing the clinical and biological data for 27 French and Belgian immunocompetent patients infected with HEV genotype 4. Their infections were probably acquired locally, since none of these patients reported traveling outside France or Belgium during the 2–8 weeks before symptoms onset. Each patient was matched for age (±5 years) and gender with two patients infected with HEV genotype 3. Bivariate analysis indicated that the HEV genotype 4-infected patients had significantly higher alanine aminotransferase (ALT) (2067 IU/L) and aspartate aminotransferase (AST) (1581 IU/L) activities and total bilirubin concentrations (92.4 μmol/L) than did those infected with HEV genotype 3 (1566 IU/L, p = 0.016; 657 IU/L, p = 0.003 and 47 μmol/L, p = 0.046) at diagnosis. In contrast, more patients infected with HEV genotype 3 reported dark urine (71% vs. 39%, p = 0.02) and experienced asthenia (89% vs. 58%, p < 0.01) than did those infected with HEV genotype 4. Two HEV genotype 4-infected patients died of multi-organ failure, while none of the genotype 3-infected patients died (p = 0.035). Finally, stepwise regression analysis retained only a greater increase in ALT (odds-ratio: 1.0005, 95% confidence interval: 1.00012–1.00084) and less frequent fever (odds-ratio = 0.1244; 95% confidence interval: 0.01887–0.82020) for patients infected with HEV genotype 4. We conclude that HEV-4 infections are likely to be associated with higher ALT activity than HEV-3 infections. Additional immunological and virological studies are required to confirm these findings and better understand the influence, if any, of genotype on HEV pathophysiology.

scholarly journals Cross-species infection of pigs with a novel rabbit, but not rat, strain of hepatitis E virus isolated in the United States

2012 ◽  
Vol 93 (8) ◽  
pp. 1687-1695 ◽  
Author(s):  
Caitlin M. Cossaboom ◽  
Laura Córdoba ◽  
Brenton J. Sanford ◽  
Pablo Piñeyro ◽  
Scott P. Kenney ◽  
...  
Keyword(s):  
United States ◽  
Capsid Protein ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
The United States ◽  
Human Pathogen ◽  
Genotype 3 ◽  
Virus Shedding ◽  
Rat Strain ◽  
Low Levels

Hepatitis E virus (HEV) is an important human pathogen. In addition to humans, HEV has also been identified in pig, chicken, mongoose, deer, rat, rabbit and fish. There are four recognized and two putative genotypes of mammalian HEV. Genotypes 1 and 2 are restricted to humans, while genotypes 3 and 4 are zoonotic. The recently identified rabbit HEV is a distant member of genotype 3. Here, we first expressed and purified the recombinant capsid protein of rabbit HEV and showed that the capsid protein of rabbit HEV cross-reacted with antibodies raised against avian, rat, swine and human HEV. Conversely, we showed that antibodies against rabbit HEV cross-reacted with capsid proteins derived from chicken, rat, swine and human HEV. Since pigs are the natural host of genotype 3 HEV, we then determined if rabbit HEV infects pigs. Twenty pigs were divided into five groups of four each and intravenously inoculated with PBS, US rabbit HEV, Chinese rabbit HEV, US rat HEV and swine HEV, respectively. Results showed that only half of the pigs inoculated with rabbit HEV had low levels of viraemia and faecal virus shedding, indicative of active but not robust HEV infection. Infection of pigs by rabbit HEV was further verified by transmission of the virus recovered from pig faeces to naïve rabbits. Pigs inoculated with rat HEV showed no evidence of infection. Preliminary results suggest that rabbit HEV is antigenically related to other HEV strains and infects pigs and that rat HEV failed to infect pigs.

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