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Hepatitis E virus lifecycle and identification of 3 forms of the ORF2 capsid protein

(2018) GASTROENTEROLOGY. 154(1). p.211-223
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Abstract
BACKGROUND & AIMS: Hepatitis E virus (HEV) infection is a major cause of acute hepatitis worldwide. Approximately 2 billion people live in areas endemic for HEV and are at risk of infection. The HEV genome encodes 3 proteins, including the ORF2 capsid protein. Detailed analyses of the HEV life cycle has been hampered by the lack of an efficient viral culture system. METHODS: We performed studies with gt3 HEV cell culture-produced particles and patient blood and stool samples. Samples were fractionated on iodixanol gradients and cushions. Infectivity assays were performed in vitro and in human liver chimeric mice. Proteins were analyzed by biochemical and proteomic approaches. Infectious particles were analyzed by transmission electron microscopy. HEV antigen levels were measured with the Wantai enzyme-linked immunosorbent assay. RESULTS: We developed an efficient cell culture system and isolated HEV particles that were infectious in vitro and in vivo. Using transmission electron microscopy, we defined the ultrastructure of HEV cell culture-produced particles and particles from patient sera and stool samples. We also identified the precise sequence of the infectious particle-associated ORF2 capsid protein. In cultured cells and in samples from patients, HEV produced 3 forms of the ORF2 capsid protein: infectious/intracellular ORF2 (ORF2i), glycosylated ORF2 (ORF2g), and cleaved ORF2 (ORF2c). The ORF2i protein associated with infectious particles, whereas the ORF2g and ORF2c proteins were massively secreted glycoproteins not associated with infectious particles. ORF2g and ORF2c were the most abundant antigens detected in sera from patients. CONCLUSIONS: We developed a cell culture system and characterized HEV particles; we identified 3 ORF2 capsid proteins (ORF2i, ORF2g, and ORFc). These findings will advance our understanding of the HEV life cycle and improve diagnosis.
Keywords
Hepatitis E, PLC/PRF/5 Cells, Infectious Particles, ORF2 Products, CHRONIC HEV INFECTION, LIVER CHIMERIC MICE, EXPRESSION, CELLS, GLYCOSYLATION, DISCOVERY, REPLICON, DEPENDS, MODEL

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MLA
Montpellier, Claire, Czeslaw Wychowski, Ibrahim Ibrahim Mahmoud Sayed, et al. “Hepatitis E Virus Lifecycle and Identification of 3 Forms of the ORF2 Capsid Protein.” GASTROENTEROLOGY 154.1 (2018): 211–223. Print.
APA
Montpellier, C., Wychowski, C., Ibrahim Mahmoud Sayed, I., Meunier, J.-C., Saliou, J.-M., Ankavay, M., Bull, A., et al. (2018). Hepatitis E virus lifecycle and identification of 3 forms of the ORF2 capsid protein. GASTROENTEROLOGY, 154(1), 211–223.
Chicago author-date
Montpellier, Claire, Czeslaw Wychowski, Ibrahim Ibrahim Mahmoud Sayed, Jean-Christophe Meunier, Jean-Michel Saliou, Maliki Ankavay, Anne Bull, et al. 2018. “Hepatitis E Virus Lifecycle and Identification of 3 Forms of the ORF2 Capsid Protein.” Gastroenterology 154 (1): 211–223.
Chicago author-date (all authors)
Montpellier, Claire, Czeslaw Wychowski, Ibrahim Ibrahim Mahmoud Sayed, Jean-Christophe Meunier, Jean-Michel Saliou, Maliki Ankavay, Anne Bull, André Pillez, Florence Abravanel, François Helle, Etienne Brochot, Hervé Drobecq, Rayan Farhat, Cécile-Marie Aliouat-Denis, Juliano G Haddad, Jacques Izopet, Philip Meuleman, Anne Goffard, Jean Dubuisson, and Laurence Cocquerel. 2018. “Hepatitis E Virus Lifecycle and Identification of 3 Forms of the ORF2 Capsid Protein.” Gastroenterology 154 (1): 211–223.
Vancouver
1.
Montpellier C, Wychowski C, Ibrahim Mahmoud Sayed I, Meunier J-C, Saliou J-M, Ankavay M, et al. Hepatitis E virus lifecycle and identification of 3 forms of the ORF2 capsid protein. GASTROENTEROLOGY. 2018;154(1):211–23.
IEEE
[1]
C. Montpellier et al., “Hepatitis E virus lifecycle and identification of 3 forms of the ORF2 capsid protein,” GASTROENTEROLOGY, vol. 154, no. 1, pp. 211–223, 2018.
@article{8548729,
  abstract     = {{BACKGROUND & AIMS: Hepatitis E virus (HEV) infection is a major cause of acute hepatitis worldwide. Approximately 2 billion people live in areas endemic for HEV and are at risk of infection. The HEV genome encodes 3 proteins, including the ORF2 capsid protein. Detailed analyses of the HEV life cycle has been hampered by the lack of an efficient viral culture system.
METHODS: We performed studies with gt3 HEV cell culture-produced particles and patient blood and stool samples. Samples were fractionated on iodixanol gradients and cushions. Infectivity assays were performed in vitro and in human liver chimeric mice. Proteins were analyzed by biochemical and proteomic approaches. Infectious particles were analyzed by transmission electron microscopy. HEV antigen levels were measured with the Wantai enzyme-linked immunosorbent assay.
RESULTS: We developed an efficient cell culture system and isolated HEV particles that were infectious in vitro and in vivo. Using transmission electron microscopy, we defined the ultrastructure of HEV cell culture-produced particles and particles from patient sera and stool samples. We also identified the precise sequence of the infectious particle-associated ORF2 capsid protein. In cultured cells and in samples from patients, HEV produced 3 forms of the ORF2 capsid protein: infectious/intracellular ORF2 (ORF2i), glycosylated ORF2 (ORF2g), and cleaved ORF2 (ORF2c). The ORF2i protein associated with infectious particles, whereas the ORF2g and ORF2c proteins were massively secreted glycoproteins not associated with infectious particles. ORF2g and ORF2c were the most abundant antigens detected in sera from patients.
CONCLUSIONS: We developed a cell culture system and characterized HEV particles; we identified 3 ORF2 capsid proteins (ORF2i, ORF2g, and ORFc). These findings will advance our understanding of the HEV life cycle and improve diagnosis.}},
  author       = {{Montpellier, Claire and Wychowski, Czeslaw and Ibrahim Mahmoud Sayed, Ibrahim and Meunier, Jean-Christophe and Saliou, Jean-Michel and Ankavay, Maliki and Bull, Anne and Pillez, André and Abravanel, Florence and Helle, François and Brochot, Etienne and Drobecq, Hervé and Farhat, Rayan and Aliouat-Denis, Cécile-Marie and Haddad, Juliano G and Izopet, Jacques and Meuleman, Philip and Goffard, Anne and Dubuisson, Jean and Cocquerel, Laurence}},
  issn         = {{0016-5085}},
  journal      = {{GASTROENTEROLOGY}},
  keywords     = {{Hepatitis E,PLC/PRF/5 Cells,Infectious Particles,ORF2 Products,CHRONIC HEV INFECTION,LIVER CHIMERIC MICE,EXPRESSION,CELLS,GLYCOSYLATION,DISCOVERY,REPLICON,DEPENDS,MODEL}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{211--223}},
  title        = {{Hepatitis E virus lifecycle and identification of 3 forms of the ORF2 capsid protein}},
  url          = {{http://dx.doi.org/10.1053/j.gastro.2017.09.020}},
  volume       = {{154}},
  year         = {{2018}},
}

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