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Mutational modifications of hepatitis A virus proteins 2B and 2C described for cell culture-adapted and attenuated virus are present in wild-type virus populations

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Abstract

Studies have identified certain mutations in the 2B and 2C proteins of hepatitis A virus (HAV) as being essential for efficient growth in cultured cells, and it is assumed that these mutations contribute to the attenuated phenotype. We found that mutations supporting cell culture growth already exist in wild-type HAV populations. This suggests that these variants are not entirely generated de novo but are selected from the wild-type population. In a prolonged case of hepatitis A, we found that sequences associated with cell culture adaptation predominated later in infection. This might suggest selection of an attenuated virus population during a prolonged clinical infection.

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References

  1. Dotzauer A (2008) Hepatitis A Virus. In: Mahy BWJ, Van Regenmortel MHV (eds) Encyclopedia of virology. Elsevier, Oxford, pp 343–350

    Chapter  Google Scholar 

  2. Dotzauer A, Kraemer L (2012) Innate and adaptive immune responses against picornaviruses and their counteractions: an overview. World J Virol 1(3):91–107

    Article  PubMed  PubMed Central  Google Scholar 

  3. Frösner GG, Deinhardt F, Scheid R, Gauss-Müller V, Holmes N, Messelberger V, Siegl G, Alexander JJ (1979) Propagation of human hepatitis A virus in a hepatoma cell line. Infection 7:303–305

    Article  PubMed  Google Scholar 

  4. Provost PJ, Hilleman MR (1979) Propagation of human hepatitis A virus in cell culture in vitro. Proc Soc Exp Biol Med 160:213–221

    Article  PubMed  CAS  Google Scholar 

  5. Flehmig B (1980) Hepatitis A-virus in cell culture: I. propagation of different hepatitis A-virus isolates in a fetal rhesus monkey kidney cell line (Frhk-4). Med Microbiol Immunol 168:239–248

    Article  PubMed  CAS  Google Scholar 

  6. Flehmig B, Vallbracht A, Wurster G (1981) Hepatitis A virus in cell culture. III. Propagation of hepatitis A virus in human embryo kidney cells and human embryo fibroblast strains. Med Microbiol Immunol 170:83–89

    Article  PubMed  CAS  Google Scholar 

  7. Gauss-Müller V, Frösner GG, Deinhardt F (1981) Propagation of hepatitis A virus in human embryo fibroblasts. J Med Virol 7:233–239

    Article  PubMed  Google Scholar 

  8. Daemer RJ, Feinstone SM, Gust ID, Purcell RH (1981) Propagation of human hepatitis A virus in African green monkey kidney cell culture: primary isolation and serial passage. Infect Immun 32:388–393

    PubMed  CAS  PubMed Central  Google Scholar 

  9. Binn LN, Lemon SM, Marchwicki RH, Redfield RR, Gates NL, Bancroft WH (1984) Primary isolation and serial passage of hepatitis A virus strains in primate cell cultures. J Clin Microbiol 20:28–33

    PubMed  CAS  PubMed Central  Google Scholar 

  10. Vallbracht A, Hofmann L, Wurster KG, Flehmig B (1984) Persistent infection of human fibroblasts by hepatitis A virus. J Gen Virol 65:609–615

    Article  PubMed  Google Scholar 

  11. De Chastonay J, Siegl G (1987) Replicative events in hepatitis A virus-infected MRC-5 cells. Virology 157:268–275

    Article  PubMed  Google Scholar 

  12. Dotzauer A, Feinstone SM, Kaplan G (1994) Susceptibility of nonprimate cell lines to hepatitis A virus infection. J Virol 68:6064–6068

    PubMed  CAS  PubMed Central  Google Scholar 

  13. Cohen JI, Rosenblum B, Ticehurst JR, Daemer RJ, Feinstone SM, Purcell RH (1987) Complete nucleotide sequence of an attenuated hepatitis A virus: comparison with wild-type virus. Proc Natl Acad Sci USA 84:2497–2501

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  14. Cohen JI, Ticehurst JR, Feinstone SM, Rosenblum B, Purcell RH (1987) Hepatitis A virus cDNA and its RNA transcripts are infectious in cell culture. J Virol 61:3035–3039

    PubMed  CAS  PubMed Central  Google Scholar 

  15. Jansen RW, Newbold JE, Lemon SM (1988) Complete nucleotide sequence of a cell culture-adapted variant of hepatitis A virus: comparison with wild-type virus with restricted capacity for in vitro replication. Virology 163:299–307

    Article  PubMed  CAS  Google Scholar 

  16. Cohen JI, Rosenblum B, Feinstone SM, Ticehurst JR, Purcell RH (1989) Attenuation and cell culture adaptation of hepatitis A virus (HAV): a genetic analysis with HAV cDNA. J Virol 63:5364–5370

    PubMed  CAS  PubMed Central  Google Scholar 

  17. Ross BC, Anderson BN, Edwards PC, Gust ID (1989) Nucleotide sequence of high-passage hepatitis A virus strain HM175: comparison with wild-type and cell culture-adapted strains. J Gen Virol 70:2805–2810

    Article  PubMed  CAS  Google Scholar 

  18. Day SP, Lemon SM (1990) A single base mutation in the 5’ noncoding region of HAV enhances replication of virus in vitro. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  19. Emerson SU, McRill C, Rosenblum B, Feinstone SM, Purcell RH (1991) Mutations responsible for adaptation of hepatitis A virus to efficient growth in cell culture. J Virol 65:4882–4886

    PubMed  CAS  PubMed Central  Google Scholar 

  20. Day SP, Murphy P, Brown EA, Lemon SM (1992) Mutations within the 5’ nontranslated region of hepatitis A virus RNA which enhance replication in BS-C-1 cells. J Virol 66:6533–6540

    PubMed  CAS  PubMed Central  Google Scholar 

  21. Emerson SU, Huang YK, McRill C, Lewis M, Purcell RH (1992) Mutations in both the 2B and 2C genes of hepatitis A virus are involved in adaptation to growth in cell culture. J Virol 66:650–654

    PubMed  CAS  PubMed Central  Google Scholar 

  22. Emerson SU, Huang YK, Purcell RH (1993) 2B and 2C mutations are essential but mutations throughout the genome of HAV contribute to adaptation to cell culture. Virology 194:475–480

    Article  PubMed  CAS  Google Scholar 

  23. Tedeschi V, Purcell RH, Emerson SU (1993) Partial characterization of hepatitis A viruses from three intermediate passage levels of a series resulting in adaptation to growth in cell culture and attenuation of virulence. J Med Virol 39:16–22

    Article  PubMed  CAS  Google Scholar 

  24. Graff J, Normann A, Feinstone SM, Flehmig B (1994) Nucleotide sequence of wild-type hepatitis A virus GBM in comparison with two cell culture-adapted variants. J Virol 68:548–554

    PubMed  CAS  PubMed Central  Google Scholar 

  25. Funkhouser AW, Purcell RH, D’Hondt E, Emerson SU (1994) Attenuated hepatitis A virus: genetic determinants of adaptation to growth in MRC-5 cells. J Virol 68:148–157

    PubMed  CAS  PubMed Central  Google Scholar 

  26. Funkhouser AW, Raychaudhuri G, Purcell RH, Govindarajan S, Elkins R, Emerson SU (1996) Progress toward the development of a genetically engineered attenuated hepatitis A virus vaccine. J Virol 70:7948–7957

    PubMed  CAS  PubMed Central  Google Scholar 

  27. Schultz DE, Honda M, Whetter LE, McKnight KL, Lemon SM (1996) Mutations within the 5’ nontranslated RNA of cell culture-adapted hepatitis A virus which enhance cap-independent translation in cultured African green monkey kidney cells. J Virol 70:1041–1049

    PubMed  CAS  PubMed Central  Google Scholar 

  28. Graff J, Normann A, Flehmig B (1997) Influence of the 5’ noncoding region of hepatitis A virus strain GBM on its growth in different cell lines. J Gen Virol 78:1841–1849

    PubMed  CAS  Google Scholar 

  29. Raychaudhuri G, Govindarajan S, Shapiro M, Purcell RH, Emerson SU (1998) Utilization of chimeras between human (HM-175) and simian (AGM-27) strains of hepatitis A virus to study the molecular basis of virulence. J Virol 72:7467–7475

    PubMed  CAS  PubMed Central  Google Scholar 

  30. Frings W, Dotzauer A (2001) Adaptation of primate cell-adapted hepatitis A virus strain HM175 to growth in guinea pig cells is independent of mutations in the 5’ nontranslated region. J Gen Virol 82:597–602

    PubMed  CAS  Google Scholar 

  31. Emerson SU, Huang YK, Nguyen H, Brockington A, Govindarajan S, St Claire M, Shapiro M, Purcell RH (2002) Identification of VP1/2A and 2C as virulence genes of hepatitis A virus and demonstration of genetic instability of 2C. J Virol 76:8551–8559

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  32. Graff J, Emerson SU (2003) Importance of amino acid 216 in nonstructural protein 2B for replication of hepatitis A virus in cell culture and in vivo. J Med Virol 71:7–17

    Article  PubMed  CAS  Google Scholar 

  33. Ticehurst JR, Racaniello VR, Baroudy BM, Baltimore D, Purcell RH, Feinstone SM (1983) Molecular cloning and characterization of hepatitis A virus cDNA. Proc Natl Acad Sci USA 80:5885–5889

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  34. Teterina NL, Bienz K, Egger D, Gorbalenya AE, Ehrenfeld E (1997) Induction of intracellular membrane rearrangements by HAV proteins 2C and 2BC. Virology 237:66–77

    Article  PubMed  CAS  Google Scholar 

  35. Jecht M, Probst C, Gauss-Müller V (1998) Membrane permeability induced by hepatitis A virus proteins 2B and 2BC and proteolytic processing of HAV 2BC. Virology 252:218–227

    Article  PubMed  CAS  Google Scholar 

  36. Gosert R, Egger D, Bienz K (2000) A cytopathic and a cell culture adapted hepatitis A virus strain differ in cell killing but not in intracellular membrane rearrangements. Virology 266:157–169

    Article  PubMed  CAS  Google Scholar 

  37. de Jong AS, de Mattia F, Van Dommelen MM, Lanke K, Melchers WJ, Willems PH, van Kuppeveld FJ (2008) Functional analysis of picornavirus 2B proteins: effects on calcium homeostasis and intracellular protein trafficking. J Virol 82:3782–3790

    Article  PubMed  PubMed Central  Google Scholar 

  38. Paulmann D, Magulski T, Schwarz R, Heitmann L, Flehmig B, Vallbracht A, Dotzauer A (2008) Hepatitis A virus protein 2B suppresses beta interferon (IFN) gene transcription by interfering with IFN regulatory factor 3 activation. J Gen Virol 89:1593–1604

    Article  PubMed  CAS  Google Scholar 

  39. Gust ID, Feinstone SM (1988) Hepatitis A. CRC Press, Boca Raton, p 95

    Google Scholar 

  40. Dotzauer A, Heitmann A, Laue T, Kraemer L, Schwabe K, Paulmann D, Flehmig B, Vallbracht A (2012) The role of immunoglobulin A in prolonged and relapsing hepatitis A virus infections. J Gen Virol 93:754–760

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Bertram Flehmig, Children´s Hospital, Department 1, University of Tübingen, Tübingen, Germany; Sigrid Baumgarte, Behörde für Soziales, Familie, Gesundheit und Verbraucherschutz, Institut für Hygiene und Umwelt, Hamburg, Germany and Meike Pahlmann, Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, for providing stool samples from hepatitis A patients. This work was supported by the Tönjes-Vagt-Stiftung, Bremen, Germany.

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Authors and Affiliations

  1. Department of Virology, University of Bremen, Leobener Straße/UFT, 28359, Bremen, Germany

    Rebecca Weilandt, Dajana Paulmann, Kore Schlottau, Angelika Vallbracht & Andreas Dotzauer

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  1. Rebecca Weilandt
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  2. Dajana Paulmann
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  3. Kore Schlottau
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  4. Angelika Vallbracht
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  5. Andreas Dotzauer
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Correspondence to Andreas Dotzauer.

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Weilandt, R., Paulmann, D., Schlottau, K. et al. Mutational modifications of hepatitis A virus proteins 2B and 2C described for cell culture-adapted and attenuated virus are present in wild-type virus populations. Arch Virol 159, 2699–2704 (2014). https://doi.org/10.1007/s00705-014-2103-6

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  • DOI: https://doi.org/10.1007/s00705-014-2103-6

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