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Hepatitis B escape mutants in Scottish blood donors

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Abstract

Hepatitis B virus (HBV) remains as the viral infection with the highest risk of transmission by transfusion. This risk is associated with window period donations, occult HBV infection (OBI) and the emergence of escape mutants, which render blood donations false negative for hepatitis B surface antigen (HBsAg) serological testing. A retrospective study was conducted to gain insights into the molecular epidemiology of HBV escape mutants in Scottish blood donors. The criterion for selection was HBV positivity either by serology or nucleic acid testing (NAT). HBsAg detection was compared across several commercial immunoassays. The full length S gene from plasma samples was PCR amplified, cloned and expressed in HepG2 cells. Eight samples showed HBsAg discordant results, while 5 OBI samples were found. Four escape mutants, containing missense mutations in the S gene, are described here. These mutations impaired HBsAg detection both from HBV infected plasma samples and from recombinant proteins derived from its infected donors. Phylogenetic analysis showed that most of the mutants were clustered in the genotype D and were closely related to strains from Asia and the Middle East. We report here a proline substitution, outside the major hydrophilic region, that impaired HBsAg detection in vivo and in vitro, warning about the risk for the emergence of vaccine escape mutants with mutations outside the major neutralisation site.

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References

  1. Brant L, Cawley C, Reynolds C, Byrne L (2008) Infection surveillance annual report 2007. The NHS Blood and Transplant and Health Protection Agency. http://www.hpa.org.uk/webc/HPAwebFile/HPAweb_C/1227255714122. Accessed 22 Nov 2012

  2. Brailsford S, Davison K, Reynolds C, Homer K, Vasconcelos M (2011) Safe supplies: new horizons. Annual Review from the NHS Blood and Transplant/Health Protection Agency. http://www.hpa.org.uk/webc/HPAwebFile/HPAweb_C/1317136707628. Accessed 22 Nov 2012

  3. Allain JP (2004) Occult hepatitis B virus infection: implications in transfusion. Vox Sang 86:83–91

    Article  PubMed  Google Scholar 

  4. Allain JP, Candotti D (2009) Diagnostic algorithm for HBV safe transfusion. Blood Transfus 7:174–182

    PubMed  Google Scholar 

  5. Allain JP, Cox L (2011) Challenges in hepatitis B detection among blood donors. Curr Opin Hematol 18:461–466

    Article  PubMed  CAS  Google Scholar 

  6. Cleland A, Nettleton P, Jarvis L, Simmonds P (1999) Use of bovine viral diarrhoea virus as an internal control for amplification of hepatitis C virus. Vox Sang 76:170–174

    Article  PubMed  CAS  Google Scholar 

  7. Jarvis L, Becker J, Tender A, Cleland A, Queiros L, Aquiar A, Azevedo J, Aprili G, Bressan F, Torres P, Nieto S, Ursitti A, Montoro J, Vila E, Ramada C, Saldanha J (2008) Evaluation of the Roche cobas s 201 system and cobas TaqScreen multiplex test for blood screening: a European multicenter study. Transfusion 48:1853–1861

    Article  PubMed  Google Scholar 

  8. Gnaneshan S, Ijaz S, Moran J, Ramsay M, Green J (2007) HepSEQ: international public health repository for hepatitis B. Nucleic Acids Res 35:D367–D370

    Article  PubMed  CAS  Google Scholar 

  9. Kay A, Zoulim F (2007) Hepatitis B virus genetic variability and evolution. Virus Res 127:164–176

    Article  PubMed  CAS  Google Scholar 

  10. Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C, Thierer T, Ashton B, Meintjes P, Drummond A (2012) Geneious basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28:1647–1649

    Article  PubMed  Google Scholar 

  11. Ponde RA (2011) The underlying mechanisms for the “isolated positivity for the hepatitis B surface antigen (HBsAg)” serological profile. Med Microbiol Immunol 200:13–22

    Article  PubMed  Google Scholar 

  12. Persson B, Argos P (1994) Prediction of transmembrane segments in proteins utilising multiple sequence alignments. J Mol Biol 237:182–192

    Article  PubMed  CAS  Google Scholar 

  13. Eble BE, MacRae DR, Lingappa VR, Ganem D (1987) Multiple topogenic sequences determine the transmembrane orientation of the hepatitis B surface antigen. Mol Cell Biol 7:3591–3601

    PubMed  CAS  Google Scholar 

  14. Blanchet M, Sureau C (2006) Analysis of the cytosolic domains of the hepatitis B virus envelope proteins for their function in viral particle assembly and infectivity. J Virol 80:11935–11945

    Article  PubMed  CAS  Google Scholar 

  15. Zuckerman JN, Zuckerman AJ (2003) Mutations of the surface protein of hepatitis B virus. Antiviral Res 60:75–78

    Article  PubMed  CAS  Google Scholar 

  16. Ma Q, Wang Y (2012) Comprehensive analysis of the prevalence of hepatitis B virus escape mutations in the major hydrophilic region of surface antigen. J Med Virol 84:198–206

    Article  PubMed  CAS  Google Scholar 

  17. Mangold CM, Streeck RE (1993) Mutational analysis of the cysteine residues in the hepatitis B virus small envelope protein. J Virol 67:4588–4597

    PubMed  CAS  Google Scholar 

  18. Huang CH, Yuan Q, Chen PJ, Zhang YL, Chen CR, Zheng QB, Yeh SH, Yu H, Xue Y, Chen YX, Liu PG, Ge SX, Zhang J, Xia NS (2012) Influence of mutations in hepatitis B virus surface protein on viral antigenicity and phenotype in occult HBV strains from blood donors. J Hepatol 57:720–729

    Article  PubMed  CAS  Google Scholar 

  19. Brojer E, Grabarczyk P, Liszewski G, Mikulska M, Allain JP, Letowska M (2006) Characterization of HBV DNA+/HBsAg− blood donors in Poland identified by triplex NAT. Hepatology 44:1666–1674

    Article  PubMed  CAS  Google Scholar 

  20. Carman WF, Van Deursen FJ, Mimms LT, Hardie D, Coppola R, Decker R, Sanders R (1997) The prevalence of surface antigen variants of hepatitis B virus in Papua New Guinea, South Africa, and Sardinia. Hepatology 26:1658–1666

    Article  PubMed  CAS  Google Scholar 

  21. Sayiner AA, Agca H, Sengonul A, Celik A, Akarsu M (2007) A new hepatitis B virus vaccine escape mutation in a renal transplant recipient. J Clin Virol 38:157–160

    Article  PubMed  CAS  Google Scholar 

  22. Hsu HY, Chang MH, Ni YH, Chiang CL, Chen HL, Wu JF, Chen PJ (2010) No increase in prevalence of hepatitis B surface antigen mutant in a population of children and adolescents who were fully covered by universal infant immunization. J Infect Dis 201:1192–1200

    Article  PubMed  Google Scholar 

  23. Norder H, Courouce AM, Coursaget P, Echevarria JM, Lee SD, Mushahwar IK, Robertson BH, Locarnini S, Magnius LO (2004) Genetic diversity of hepatitis B virus strains derived worldwide: genotypes, subgenotypes, and HBsAg subtypes. Intervirology 47:289–309

    Article  PubMed  Google Scholar 

  24. Salisse J, Sureau C (2009) A function essential to viral entry underlies the hepatitis B virus “a” determinant. J Virol 83:9321–9328

    Article  PubMed  CAS  Google Scholar 

  25. Carman WF, Zanetti AR, Karayiannis P, Waters J, Manzillo G, Tanzi E, Zuckerman AJ, Thomas HC (1990) Vaccine-induced escape mutant of hepatitis B virus. Lancet 336:325–329

    Article  PubMed  CAS  Google Scholar 

  26. Kalinina T, Iwanski A, Will H, Sterneck M (2003) Deficiency in virion secretion and decreased stability of the hepatitis B virus immune escape mutant G145R. Hepatology 38:1274–1281

    Article  PubMed  CAS  Google Scholar 

  27. Avellon A, Echevarria JM (2006) Frequency of hepatitis B virus ‘a’ determinant variants in unselected Spanish chronic carriers. J Med Virol 78:24–36

    Article  PubMed  Google Scholar 

  28. Chua PK, Wang RY, Lin MH, Masuda T, Suk FM, Shih C (2005) Reduced secretion of virions and hepatitis B virus (HBV) surface antigen of a naturally occurring HBV variant correlates with the accumulation of the small S envelope protein in the endoplasmic reticulum and Golgi apparatus. J Virol 79:13483–13496

    Article  PubMed  CAS  Google Scholar 

  29. Ito K, Qin Y, Guarnieri M, Garcia T, Kwei K, Mizokami M, Zhang J, Li J, Wands JR, Tong S (2010) Impairment of hepatitis B virus virion secretion by single-amino-acid substitutions in the small envelope protein and rescue by a novel glycosylation site. J Virol 84:12850–12861

    Article  PubMed  CAS  Google Scholar 

  30. Warner N, Locarnini S (2008) The antiviral drug selected hepatitis B virus rtA181T/sW172* mutant has a dominant negative secretion defect and alters the typical profile of viral rebound. Hepatology 48:88–98

    Article  PubMed  CAS  Google Scholar 

  31. Clements CJ, Coghlan B, Creati M, Locarnini S, Tedder RS, Torresi J (2010) Global control of hepatitis B virus: does treatment-induced antigenic change affect immunization? Bull World Health Organ 88:66–73

    Article  PubMed  Google Scholar 

  32. Scheiblauer H, El-Nageh M, Diaz S, Nick S, Zeichhardt H, Grunert HP, Prince A (2010) Performance evaluation of 70 hepatitis B virus (HBV) surface antigen (HBsAg) assays from around the world by a geographically diverse panel with an array of HBV genotypes and HBsAg subtypes. Vox Sang 98:403–414

    Article  PubMed  CAS  Google Scholar 

  33. Bar-Yishay I, Shaul Y, Shlomai A (2011) Hepatocyte metabolic signalling pathways and regulation of hepatitis B virus expression. Liver Int 31:282–290

    Article  PubMed  CAS  Google Scholar 

  34. Chandra PK, Biswas A, Datta S, Banerjee A, Panigrahi R, Chakrabarti S, De BK, Chakravarty R (2009) Subgenotypes of hepatitis B virus genotype D (D1, D2, D3 and D5) in India: differential pattern of mutations, liver injury and occult HBV infection. J Viral Hepat 16:749–756

    Article  PubMed  CAS  Google Scholar 

  35. Barclay ST, Cameron S, Mills PR, Priest M, Ross F, Fox R, Goulding C, Forrest EH, Morris AJ, Neilson M, Stanley AJ (2010) The changing face of hepatitis B in greater Glasgow: epidemiological trends 1993–2007. Scott Med J 55:4–7

    Article  PubMed  CAS  Google Scholar 

  36. Mackenzie AR, Molyneaux PJ, Cadwgan AM, Laing RB, Douglas JG, Smith CC (2003) Increasing incidence of acute hepatitis B virus infection referrals to the Aberdeen Infection Unit: a matter for concern. Scott Med J 48:73–75

    PubMed  CAS  Google Scholar 

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Acknowledgments

We are very thankful to Helen Munro and Tony Jordan for providing serology results, Jacqui Doran for HBV NAT and all SNBTS teams involved in sample collection and storage.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Microbiology and Components Group, RDI, Scottish National Blood Transfusion Service/NHS, 21 Ellen’s Glen Road, Edinburgh, EH17 7QT, UK

    Osmany Larralde, Brian Dow, Lisa Jarvis, Fiona Davidson & Juraj Petrik

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  1. Osmany Larralde
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  2. Brian Dow
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  3. Lisa Jarvis
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  5. Juraj Petrik
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Correspondence to Osmany Larralde.

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Larralde, O., Dow, B., Jarvis, L. et al. Hepatitis B escape mutants in Scottish blood donors. Med Microbiol Immunol 202, 207–214 (2013). https://doi.org/10.1007/s00430-012-0283-9

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  • DOI: https://doi.org/10.1007/s00430-012-0283-9

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