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Expression and Immunoreactivities of Hepatitis E Virus Genotype 3 Open Reading Frame-2 (ORF-2) Recombinant Proteins Expressed in Insect Cells

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Abstract

Hepatitis E virus (HEV) is a fecal-orally transmitted virus that is endemic in many geographical areas with poor sanitary conditions and inadequate water supplies. In Europe, a low-endemic area, an increased number of autochthonous sporadic human cases of patients infected with HEV strains of genotype 3, have been reported lately. The relatively high prevalence of HEV genotype 3 infections in European pigs has raised concerns about a potential zoonotic transmission to humans. Determination of HEV seroprevalence in pigs would help to clarify its incidence and possible zoonotic implications. To this purpose, we have expressed and partially characterized swine genotype 3 HEV open reading frame-2 proteins upon infection of Sf21 insect cells with recombinant baculoviruses. The use of the expressed proteins as diagnostic antigens for the detection of antibodies to HEV has been further assayed with human and swine sera.

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References

  • Arankalle, V. A., Lole, K. S., Deshmukh, T. M., Chobe, L. P., & Gandhe, S. S. (2007). Evaluation of human (genotype 1) and swine (genotype 4)-ORF2-based ELISAs for anti-HEV IgM and IgG detection in an endemic country and search for type 4 human HEV infections. Journal of Viral Hepatitis, 14(6), 435–445.

    Article  PubMed  CAS  Google Scholar 

  • Banks, M., Heath, G. S., Grieson, S. S., King, D. P., Gresahm, A., Gironés, R., et al. (2004). Evidence for the presence of hepatitis E virus in pigs in the United Kingdom. Veterinary Records, 154(8), 223–227.

    CAS  Google Scholar 

  • Borgen, K, Herremans, T., Duizer, E., Vennema, H., Rutjes, S., Bosman, A., et al. (2008). Non-travel related hepatitis E virus genotype 3 infections in the Netherlands: A case series 2004–2006. BMC Infectious Diseases. doi:10.1186/1471-2334-8-61.

  • Bouwknegt, M., Engel, B., Herremans, M. M. P. T., Widdowson, M. A., Worm, H. C., Koopmans, M. P. G., et al. (2007a). Bayesian estimation of Hepatitis E virus seroprevalence for populations with different exposure levels to swine in the Netherlands. Epidemiology and Infection, 136(4), 567–576.

    PubMed  Google Scholar 

  • Bouwknegt, M., Lodder-Verschoor, F., van der Poel, W. H., Rutjes, S. A., & de Roda Husman, A. M. (2007b). Hepatitis E virus RNA in commercial porcine livers in the Netherlands. Journal of Food Protection, 70(12), 2889–2895.

    PubMed  CAS  Google Scholar 

  • Bouwknegt, M., Frankena, K., Rutjes, S. A., Wellenberg, G. J., de Roda Husman, A. M., van der Poel, W. H., et al. (2008). Estimation of hepatitis E virus transmission among pigs due to contact exposure. Veterinary Research. doi:10.1051/vetres:2008017.

  • Buti, M., Clemente-Casares, P., Jardi, R., Formiga-Cruz, M., Schaper, M., Valdes, A., et al. (2004). Sporadic cases of acute autochthonous hepatitis E in Spain. Journal of Hepatology, 41(1), 126–131.

    Article  PubMed  Google Scholar 

  • Buti, M., Dominguez, A., Plans, P., Jardi, R., Schaper, M., Espuñes, J., et al. (2006). Community-based seroepidemiological survey of hepatitis E virus infection in Catalonia, Spain. Clinical and Vaccine Immunology, 13(21), 1328–1332.

    Article  PubMed  CAS  Google Scholar 

  • Casas, M., Pujols, J., Rosell, R., de Deus, N, Peralta, B., Pina, S., et al. (2008). Retrospective serological study of hepatitis E infection in pigs from 1985 to 1997 in Spain. Veterinary Microbiology. doi:10.1016/j.vetmic.2008.09.075.

  • Dalton, H. R., Stableforth, W., Thurairajah, P., Hazeldine, S., Remnarace, R., Usama, W., et al. (2008). Autochthonous hepatitis E in Southwest England: Natural history, complications and seasonal variation, and hepatitis E virus IgG seroprevalence in blood donors, the elderly and patients with chronic liver disease. European Journal of Gastroenterology and Hepatology, 20(8), 784–790.

    Article  PubMed  CAS  Google Scholar 

  • Daniel, H. D., Warier, A., Abraham, P., & Sridharan, G. (2004). Age-wise exposure rates to hepatitis E virus in Southern Indian patient population without liver disease. The American Journal of Tropical Medicine and Hygiene, 71(5), 675–678.

    PubMed  Google Scholar 

  • De Deus, N., Casas, M., Peralta, B., Nofrarias, M., Pina, S., Martín, M., et al. (2008). Hepatitis E virus infection dynamics and organic distribution in naturally infected pigs in a farrow to finish farm. Veterinary Microbiology, 132(1–2), 19–28.

    Article  PubMed  CAS  Google Scholar 

  • Drobeniuc, J., Favorov, M. O., Shapiro, C. N., Bell, B. P., Mast, E. E., Dadu, A., et al. (2001). Hepatitis E virus antibody prevalence among persons who work with swine. Journal of Infectious Diseases, 184(12), 1594–1597.

    Article  PubMed  CAS  Google Scholar 

  • Feagins, A. R., Opriessnig, T., Guenette, D. K., Halbur, P. G., & Meng, X. J. (2007). Detection and characterization of infectious hepatitis E virus from commercial pig livers sold in local grocery stores in the USA. Journal of General Virology, 88(3), 912–917.

    Article  PubMed  CAS  Google Scholar 

  • Fernández-Barredo, S., Galiana, C., García, A., Vega, S., Gómez, M. T., & Pérez-Gracia, M. T. (2006). Detection of hepatitis E virus shedding in feces of pigs at different stages of production using reverse transcription polymerase chain reaction. The Journal of Veterinary Diagnostic Investigation, 18(5), 462–465.

    Google Scholar 

  • Galiana, C., Fernández-Barredo, S., García, A., Gómez, M. T., & Pérez-Gracia, M. T. (2008). Occupational exposure to hepatitis E virus (HEV) in swine workers. The American Journal of Tropical Medicine and Hygiene, 78(6), 1012–1015.

    PubMed  Google Scholar 

  • He, J., Tam, A. W., Yarbough, P. O., Reyes, R. G., & Carl, M. (1993). Expression and diagnostic utility of hepatitis E virus putative structural proteins expressed in insect cells. Journal of Clinical Microbiology, 31(8), 2167–2173.

    PubMed  CAS  Google Scholar 

  • He, J., Ching, W. M., Yarbough, P., Wang, H., & Carl, M. (1995). Purification of a baculovirus-expressed hepatitis E virus structural protein and utility in an enzyme-linked immunosorbent assay. Journal of Clinical Microbiology, 33(12), 3308–3311.

    PubMed  CAS  Google Scholar 

  • Herremans, M., Baker, J., Duizer, E., Vennema, H., & Koopmans, M. P. (2007a). Use of serological assays for diagnosis of hepatitis E virus genotype 1 and 3 infections in a setting of low endemicity. Clinical and Vaccine Immunology, 14(5), 562–568.

    Article  PubMed  CAS  Google Scholar 

  • Herremans, M., Duizer, E., Jusic, E., & Koopmans, M. P. (2007b). Detection of hepatitis E virus-specific immunoglobulin in a patient infected with hepatitis E virus genotype 1 or 3. Clinical and Vaccine Immunology, 14(3), 276–280.

    Article  PubMed  CAS  Google Scholar 

  • Li, F., Torresi, J., Locarnini, S. A., Zhuang, H., Zhu, W., Guo, X., et al. (1997a). Amino-terminal epitopes are exposed when full-length open reading frame 2 of hepatitis E virus is expressed in Escherichia coli, but carboxy-terminal epitopes are masked. Journal Medical Virology, 52(3), 289–300.

    Article  CAS  Google Scholar 

  • Li, T. C., Yamakawa, Y., Suzuki, K., Tatsumi, M., Razak, M. A., Uchida, T., et al. (1997b). Expression and self-assembly of empty virus-like particles of hepatitis E virus. Journal of Virology, 71(10), 7207–7213.

    PubMed  CAS  Google Scholar 

  • Mansuy, J. M., Peron, J. M., Abravanel, F., Poirson, H., Dubois, M., Miedouge, M., et al. (2004). Hepatitis E in South West France in individuals who have never visited an endemic area. Journal of Medical Virology, 74(3), 419–424.

    Article  PubMed  Google Scholar 

  • Matsuda, H., Okada, K., Takahasi, K., & Mishiro, S. (2003). Severe hepatitis E virus infection after ingestion of uncooked liver from a wild boar. Journal of Infectious Diseases, 188(6), 944.

    Article  PubMed  Google Scholar 

  • Meng, X. J. (2003). Swine hepatitis E virus: Cross-species infection and risk in xenotransplantation. Current Topics in Microbiology and Immunology, 278, 185–216.

    PubMed  CAS  Google Scholar 

  • Meng, X. J., Wiseman, B., Elvinger, F., Guenette, D. K., Toth, T. E., Engle, R. E., et al. (2002). Prevalence of antibodies to hepatitis E virus in veterinarians working with swine and in normal blood donors in the United States and other countries. Journal of Clinical Microbiology, 40(1), 117–122.

    Article  PubMed  CAS  Google Scholar 

  • Mizuo, H., Yazaki, Y., Sugarawa, K., Tsuda, F., Takahasi, M., Nishizawa, T., et al. (2005). Possible risk factors for the transmission of hepatitis E virus and for the severe form of hepatitis E acquired locally in Hokkaido, Japan. Journal of Medical Virology, 76(3), 341–349.

    Article  PubMed  CAS  Google Scholar 

  • Olsen, B., Axelsson-Olsson, D., Thelin, A., & Weiland, O. (2006). Unexpected high prevalence of IgG-antibodies to hepatitis E virus in Swedish pig farmers and controls. The Scandinavian Journal of Infectious Diseases, 38(1), 55–58.

    Article  Google Scholar 

  • Peréz-Gracia, M. T., García-Valdivia, M. S., Galán, F., & Rodríguez-Iglesias, M. A. (2004). Detection of hepatitis E virus in patients sera in southern Spain. Acta Virologica, 48(3), 197–200.

    PubMed  Google Scholar 

  • Pintó, R., & Saiz, J. C. (2007). Human viruses in water. In A. Bosch (Ed.), Perspectives in medical virology (Vol. 17, pp. 39–67). Amsterdam, The Netherlands: Elsevier.

    Google Scholar 

  • Preiss, J. C., Plentz, A., Engelman, E., Schneider, T., Jilg, W., Zeitz, M., et al. (2006). Autochthonous hepatitis E virus infection in Germany with sequence similarities to other European isolates. Infection, 34(3), 173–175.

    Article  PubMed  CAS  Google Scholar 

  • Robinson, R. A., Burgess, W. H., Emerson, S. U., Leibowitz, R. S., Sosnovtseva, S. A., Tsarev, S., et al. (1998). Structural characterization of recombinant hepatitis E virus ORF2 proteins in baculovirus-infected insect cells. Protein Expression and Purification, 12(1), 75–84.

    Article  PubMed  CAS  Google Scholar 

  • Seminati, C., Mateu, E., Peralta, B., de Deus, N., & Martín, M. (2008). Distribution of hepatitis E virus infection and its prevalence in pigs on commercial farms in Spain. Veterinary Journal, 175(1), 130–132.

    Article  CAS  Google Scholar 

  • Tam, A. W., Smith, M. M., Guerra, M. E., Huang, C. C., Bradley, D. W., Fry, K. E., et al. (1991). Hepatitis E virus (HEV): molecular cloning and sequencing of the full-length viral genome. Virology, 185(1), 120–131.

    Article  PubMed  CAS  Google Scholar 

  • Tei, S., Kitajimam, N., Takahashim, K., & Mishirom, S. (2003). Zoonotic transmission of hepatitis E virus from deer to human beings. The Lancet, 362(9381), 371–373.

    Article  Google Scholar 

  • Tsarev, S. A., Tsareva, T. S., Emerson, S. U., Kapikian, A. Z., Ticehurst, J., London, W., et al. (1993). ELISA for antibody to hepatitis E virus (HEV) based on complete open-reading frame 2 protein expressed in insect cells: Identification of HEV infection in primates. Journal of Infectious Diseases, 168(2), 369–378.

    PubMed  CAS  Google Scholar 

  • Van der Poel, W. H., Verschoor, F., van der Heide, R., Herrera, M. I., Vivo, A., Kooreman, M., et al. (2001). Hepatitis E virus sequences in swine related to sequences in humans, The Netherlands. Emerging Infectious Diseases, 7(6), 970–976.

    Article  PubMed  Google Scholar 

  • Wang, Y., Zhang, H., Li, Z., Gu, W., Lan, H., Hao, W., et al. (2001). Detection of sporadic cases of hepatitis E virus (HEV) infection in China using immunoassays based on recombinant open reading frame 2 and 3 polypeptides from HEV genotype 4. Journal of Clinical Microbiology, 39(12), 4370–4379.

    Article  PubMed  CAS  Google Scholar 

  • Widdowson, M. A., Jaspers, W. J., van der Poel, W. H., Verschoor, F., de Roda Husman, A. M., Winter, H. L., et al. (2003). Cluster of cases of acute hepatitis associated with hepatitis E virus infection acquired in the Netherlands. Clinical Infectious Diseases, 36(1), 29–33.

    Article  PubMed  Google Scholar 

  • Withers, M. R., Correa, M. T., Morrow, M., Stebbins, M. E., Seriwatana, J., Webster, W. D., et al. (2002). Antibody levels to hepatitis E virus in North Carolina swine workers, non-swine workers, swine, and murids. The American Journal of Tropical Medicine & Hygiene, 66(4), 384–388.

    Google Scholar 

  • Worm, H. C., van der Poel, W. H. M., & Brandstätter, G. (2002). Hepatitis E: An overview. Microbes & Infection, 4(8), 657–666.

    Article  CAS  Google Scholar 

  • Yazaki, Y., Mizuo, H., Takahasi, M., Nishizawa, T., Sasaki, N., Goatnada, Y., et al. (2003). Sporadic acute hepatitis E in Hokkaiso, Japan, may be food-borne, as suggested by the presence of hepatitis E virus in pig liver as food. Journal of General Virology, 84(9), 2351–2357.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, Y., McAtee, P., Yarbourgh, P. O., Tam, A. W., & Fuerst, T. (1997). Expression, characterization, and immunoreactivities of a soluble hepatitis E virus putative capsid protein species expressed in insect cells. Clinical Diagnostic Laboratory Immunology, 4(4), 423–428.

    CAS  Google Scholar 

  • Zhang, J. Z., Im, S. W., Lau, S. H., Cahu, T. N., Lai, S. T., Ng, S. P., et al. (2002). Occurrence of hepatitis E virus IgM, low avidity IgG serum antibodies, and viremia in sporadic cases of non-A,-B,-C acute hepatitis. Journal of Medical Virology, 66(1), 40–48.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The study was supported in part by grants (AGL2004-06071 and CSD2006-0007) from the Spanish Ministerio de Ciencia e Innovación (MICINN) to J.C.S. N.J.O. has been supported by a scholarship from Instituto Nacional de Investigación y Tecnología Agraraia y Alimentaria (INIA) and by a COST-929 short-term mission. J.A. was supported by a scholarship from the MICIIN. E.E.R. was supported by a contract ascribed to grant CSD2006-0007 from MICIIN. We are indebted to M.T. Pérez-Gracia (U. Cardenal Herrera-CEU, Valencia, Spain), M. Buti (Hospital Vall d′Hebron. Barcelona, Spain), and M. Martín (CReSA, UAB, Barcelona, Spain) for kindly providing us with swine fecal extract, human and swine sera, respectively.

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Authors and Affiliations

  1. Department of Biotechnology, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra. Coruña km. 7.5, 28040, Madrid, Spain

    Nereida Jiménez de Oya, Inmaculada Galindo, Estela Escribano-Romero, Ana-Belén Blázquez, Julio Alonso-Padilla, José M. Escribano & Juan-Carlos Saiz

  2. Department of Infectious Pathology and Epidemiology, Faculty of Veterinaria, University of Zaragoza, Zaragoza, Spain

    Nabil Halaihel

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  1. Nereida Jiménez de Oya
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  2. Inmaculada Galindo
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  3. Estela Escribano-Romero
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  4. Ana-Belén Blázquez
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  5. Julio Alonso-Padilla
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  6. Nabil Halaihel
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  7. José M. Escribano
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  8. Juan-Carlos Saiz
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Correspondence to Juan-Carlos Saiz.

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Jiménez de Oya, N., Galindo, I., Escribano-Romero, E. et al. Expression and Immunoreactivities of Hepatitis E Virus Genotype 3 Open Reading Frame-2 (ORF-2) Recombinant Proteins Expressed in Insect Cells. Food Environ Virol 1, 77–84 (2009). https://doi.org/10.1007/s12560-009-9012-y

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