1887

Abstract

Occult hepatitis B is defined by the presence of hepatitis B virus (HBV) DNA in the absence of hepatitis B surface antigen (HBsAg). Occult HBV is associated with the development of hepatocellular carcinoma, reactivation during immune suppression, and virus transmission. Viral mutations contribute significantly to the occult HBV phenotype. Mutations in the ‘a’ determinant of HBsAg are of particular interest, as these mutations are associated with immune escape, vaccine escape and diagnostic failure. We examined the effects of selected occult HBV-associated mutations identified in a population of HIV-positive South Africans on HBsAg production in vitro. Mutations were inserted into two different chronic HBV backbones and transfected into a hepatocyte-derived cell line. HBsAg levels were quantified by enzyme-linked immunosorbent assay (ELISA), while the detectability of mutant HBsAg was determined using an HA-tagged HBsAg expression system. Of the seven mutations analysed, four (S132P, C138Y, N146D and C147Y) resulted in decreased HBsAg expression in one viral background but not in the second viral background. One mutation (N146D) led to a decrease in HBsAg detected as compared to HA-tag, indicating that this mutation compromises the ability of the ELISA to detect HBsAg. The contribution of occult-associated mutations to the HBsAg-negative phenotype of occult HBV cannot be determined adequately by testing the effect of the mutation in a single viral background, and rigorous analysis of these mutations is required.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.000469
2016-07-01
2019-10-13
Loading full text...

Full text loading...

/deliver/fulltext/jgv/97/7/1615.html?itemId=/content/journal/jgv/10.1099/jgv.0.000469&mimeType=html&fmt=ahah

References

  1. Ahmadabadi B. N., Hassanshahi G., Arababadi M. K., Leanza C., Kennedy D..( 2012;). The IL-10 promoter polymorphism at position -592 is correlated with susceptibility to occult HBV infection. . Inflammation 35: 818–821. [CrossRef] [PubMed]
    [Google Scholar]
  2. Arababadi M. K., Pourfathollah A. A., Jafarzadeh A., Hassanshahi G., Rezvani M. E..( 2010;). Association of exon 9 but not intron 8 VDR polymorphisms with occult HBV infection in south-eastern iranian patients. . J Gastroenterol Hepatol 25: 90–93. [CrossRef] [PubMed]
    [Google Scholar]
  3. Araujo N. M., Branco-Vieira M., Silva A. C., Pilotto J. H., Grinsztejn B., de Almeida A. J., Trepo C., Gomes S. A..( 2008;). Occult hepatitis B virus infection in hiv-infected patients: Evaluation of biochemical, virological and molecular parameters. . Hepatol Res 38: 1194–1203. [CrossRef] [PubMed]
    [Google Scholar]
  4. Bell T. G., Makondo E., Martinson N. A., Kramvis A..( 2012;). Hepatitis B virus infection in human immunodeficiency virus infected southern african adults: Occult or overt – that is the question. . PLoS One 7: e45750. [CrossRef] [PubMed]
    [Google Scholar]
  5. Biswas S., Candotti D., Allain J. P..( 2013;). Specific amino acid substitutions in the S protein prevent its excretion in vitro and may contribute to occult hepatitis B virus infection. . J Virol 87: 7882–7892. [CrossRef] [PubMed]
    [Google Scholar]
  6. Bruni R., Prosperi M., Marcantonio C., Amadori A., Villano U., Tritarelli E., Presti A., Ciccozzi M., Ciccaglione A. R..( 2011;). A computational approach to identify point mutations associated with occult hepatitis B: Significant mutations affect coding regions but not regulative elements of HBV. . Virol J 8: 394. BioMed Central Ltd. [CrossRef] [PubMed]
    [Google Scholar]
  7. Cacciola I., Pollicino T., Squadrito G., Cerenzia G., Orlando M. E., Raimondo G..( 1999;). Occult hepatitis B virus infection in patients with chronic hepatitis C liver disease. . N Engl J Med 341: 22–26. [CrossRef] [PubMed]
    [Google Scholar]
  8. Candotti D., Opare-Sem O., Rezvan H., Sarkodie F., Allain J. P..( 2006;). Molecular and serological characterization of hepatitis B virus in deferred ghanaian blood donors with and without elevated alanine aminotransferase. . J Viral Hepat 13: 715–724. [CrossRef] [PubMed]
    [Google Scholar]
  9. Carman W. F..( 1997;). The clinical significance of surface antigen variants of hepatitis B virus. . J Viral Hepat 4: Suppl 1 11–20. [CrossRef] [PubMed]
    [Google Scholar]
  10. Carman W. F., Zanetti A. R., Karayiannis P., Waters J., Manzillo G., Tanzi E., Zuckerman A. J., Thomas H. C..( 1990;). Vaccine-induced escape mutant of hepatitis B virus. . Lancet (London, England) 336: 325–329. [CrossRef] [PubMed]
    [Google Scholar]
  11. Cavallone D., Moriconi F., Colombatto P., Oliveri F., Bonino F., Brunetto M. R..( 2013;). Optimization of in vitro HBV replication and HBsAg production in HuH7 cell line. . J Virol Methods 189: 110–117. Elsevier B.V. [CrossRef] [PubMed]
    [Google Scholar]
  12. Chaudhuri V., Tayal R., Nayak B., Acharya S. K., Panda S. K..( 2004;). Occult hepatitis B virus infection in chronic liver disease: Full-length genome and analysis of mutant surface promoter. . Gastroenterology 127: 1356–1371. [CrossRef] [PubMed]
    [Google Scholar]
  13. Chazouillères O., Mamish D., Kim M., Carey K., Ferrell L., Roberts J. P., Ascher N. L., Wright T. L..( 1994;). "Occult" hepatitis B virus as source of infection in liver transplant recipients. . Lancet (London) 343: 142–146. [CrossRef] [PubMed]
    [Google Scholar]
  14. Chen C. Y., Hsu H. Y., Liu C. C., Chang M. H., Ni Y. H..( 2010;). Stable seroepidemiology of hepatitis B after universal immunization in taiwan: A 3-year study of national surveillance of primary school students. . Vaccine 28: 5605–5608. Elsevier Ltd. [CrossRef] [PubMed]
    [Google Scholar]
  15. Chen C. H., Changchien C. S., Lee C. M., Tung W. C., Hung C. H., Hu T. H., Wang J. H., Wang J. C., Lu S.-N..( 2009;). A study on sequence variations in pre-S/surface, X and enhancer ii/core promoter/precore regions of occult hepatitis B virus in non-B, non-C hepatocellular carcinoma patients in Taiwan. . Int J Cancer 125: 621–629. [CrossRef] [PubMed]
    [Google Scholar]
  16. Chong-Jin O., Wei Ning C., Shiuan K., Gek Keow L..( 1999;). Identification of hepatitis B surface antigen variants with alterations outside the "a" determinant in immunized singapore infants. . J Infect Dis 179: 259–263. [CrossRef] [PubMed]
    [Google Scholar]
  17. Coleman P. F., Chen Y. C., Mushahwar I. K..( 1999;). Immunoassay detection of hepatitis B surface antigen mutants. . J Med Virol 59: 19–24. [CrossRef] [PubMed]
    [Google Scholar]
  18. Coppola N., Tonziello G., Pisaturo M., Messina V., Guastafierro S., Fiore M., Iodice V., Sagnelli C., Stanzione M. et al.( 2011;). Reactivation of overt and occult hepatitis B infection in various immunosuppressive settings. . J Med Virol 83: 1909–1916. [CrossRef] [PubMed]
    [Google Scholar]
  19. Cui X. J., Cho Y. K., Song B. C..( 2015;). Influence of the basal core promoter and precore mutation on replication of hepatitis B virus and antiviral susceptibility of different genotypes. . J Med Virol 87: 601–608. [CrossRef] [PubMed]
    [Google Scholar]
  20. Donlin M. J., Szeto B., Gohara D. W., Aurora R., Tavis J. E..( 2012;). Genome-wide networks of amino acid covariances are common among viruses. . J Virol 86: 3050–3063. [CrossRef] [PubMed]
    [Google Scholar]
  21. Firnhaber C., Viana R., Reyneke A., Schultze D., Malope B., Maskew M., Di Bisceglie A., MacPhail P., Sanne I., Kew M..( 2011;). Occult hepatitis B virus infection in patients with isolated core antibody and HIV co-infection in an urban clinic in Johannesburg, South Africa. . Int J Infect Dis 13: 488–492. [CrossRef] [PubMed]
    [Google Scholar]
  22. Ghaziasadi A., Alavian S. M., Norouzi M., Fazeli Z., Jazayeri S. M..( 2013;). Mutational analysis of HBs Ag-Positive mothers and their infected children despite immunoprophylaxis. . Iran J Allergy Asthma Immunol 12: 352–360.[PubMed]
    [Google Scholar]
  23. Günther S., Li B. C., Miska S., Krüger D. H., Meisel H., Will H..( 1995;). A novel method for efficient amplification of whole hepatitis B virus genomes permits rapid functional analysis and reveals deletion mutants in immunosuppressed patients. . J Virol 69: 5437–5444.[PubMed]
    [Google Scholar]
  24. Hass M., Hannoun C., Kalinina T., Sommer G., Manegold C., Günther S..( 2005;). Functional analysis of hepatitis B virus reactivating in hepatitis B surface antigen-negative individuals. . Hepatology 42: 93–103. [CrossRef] [PubMed]
    [Google Scholar]
  25. Hassanshahi G., Arababadi M. K., Khoramdelazad H., Yaghini N., Zarandi E. R..( 2010;). Assessment of CXCL12 (sdf-1alpha) polymorphisms and its serum level in posttransfusion occult hbv-infected patients in Southeastern Iran. . Arch Med Res 41: 338–342. [CrossRef] [PubMed]
    [Google Scholar]
  26. He C., Nomura F., Itoga S., Isobe K., Nakai T..( 2001;). Prevalence of vaccine-induced escape mutants of hepatitis B virus in the adult population in china: A prospective study in 176 restaurant employees. . J Gastroenterol Hepatol 16: 1373–1377. [CrossRef] [PubMed]
    [Google Scholar]
  27. Hollinger F. B., Sood G..( 2010;). Occult hepatitis B virus infection: A covert operation. . J Viral Hepat 17: 1–15. [CrossRef] [PubMed]
    [Google Scholar]
  28. Hsu H. Y., Chang M. H., Liaw S. H., Ni Y. H., Chen H. L..( 1999;). Changes of hepatitis B surface antigen variants in carrier children before and after universal vaccination in taiwan. . Hepatology 30: 1312–1317. [CrossRef] [PubMed]
    [Google Scholar]
  29. Huang C. H., Yuan Q., Chen P. J., Zhang Y. L., Chen C. R., Zheng Q. B., Yeh S. H., Yu H., Xue Y. et al.( 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. [CrossRef] [PubMed]
    [Google Scholar]
  30. Hui C. K., Cheung W. W., Zhang H. Y., Au W. Y., Yueng Y. H., Leung A. Y., Leung N., Luk J. M., Lie A. K..( 2006;). Kinetics and risk of de novo hepatitis B infection in hbsag-negative patients undergoing cytotoxic chemotherapy. . Gastroenterology 131: 59–68. [CrossRef] [PubMed]
    [Google Scholar]
  31. Ikeda K., Kobayashi M., Someya T., Saitoh S., Hosaka T., Akuta N., Suzuki F., Suzuki Y., Arase Y., Kumada H..( 2009;). Occult hepatitis B virus infection increases hepatocellular carcinogenesis by eight times in patients with non-B, non-C liver cirrhosis: A cohort study. . J Viral Hepat 16: 437–443. [CrossRef] [PubMed]
    [Google Scholar]
  32. Ireland J. H., O'Donnell B., Basuni A. A., Kean J. D., Wallace L. A., Lau G. K., Carman W. F..( 2000;). Reactivity of 13 in vitro expressed hepatitis B surface antigen variants in 7 commercial diagnostic assays. . Hepatology 31: 1176–1182. [CrossRef] [PubMed]
    [Google Scholar]
  33. Ishigami M., Honda T., Ishizu Y., Onishi Y., Kamei H., Hayashi K., Ogura Y., Hirooka Y., Goto H..( 2007;). Frequent incidence of escape mutants after successful hepatitis B vaccine response and stopping of nucleos(t)ide analogues in liver transplant recipients. . Liver Transplant 13: 465–466.
    [Google Scholar]
  34. Jammeh S., Thomas H. C., Karayiannis P..( 2007;). Replicative competence of the T131I, K141E, and G145R surface variants of hepatitis B virus. . J Infect Dis 196: 1010–1013. [CrossRef] [PubMed]
    [Google Scholar]
  35. Kao J. H., Chen P. J., Lai M. Y., Chen D.-S..( 2002;). Occult hepatitis B virus infection and clinical outcomes of patients with chronic hepatitis C. . J Clin Microbiol 40: 4068–4071. [CrossRef] [PubMed]
    [Google Scholar]
  36. Kew M. C., Welschinger R., Viana R..( 2008;). Occult hepatitis B virus infection in Southern African blacks with hepatocellular carcinoma. . J Gastroenterol Hepatol 23: 1426–1430. [CrossRef] [PubMed]
    [Google Scholar]
  37. Kew M., Van Damme P., Mphahlele, M. J..( 2001;). Mutant hepatitis B viruses: A matter of academic interest only or a problem with far-reaching implications? Guido franc. . Hepatol Res 19: 3799–3815.
    [Google Scholar]
  38. Lee K. M., Kim Y. S., Ko Y. Y., Yoo B. M., Lee K. J., Kim J. H., Hahm K. B., Cho S. W..( 2001;). Emergence of vaccine-induced escape mutant of hepatitis B virus with multiple surface gene mutations in a Korean child. . J Korean Med Sci 16: 359–362. [CrossRef] [PubMed]
    [Google Scholar]
  39. Lee L. Y., Aw M., Raugg M., Loh K. S., Lim S. G., Lee G. H..( 2015;). Hepatitis B immunoprophylaxis failure and the presence of hepatitis B surface gene mutants in the affected children. . J Med Virol 87: 1344–1350. [CrossRef] [PubMed]
    [Google Scholar]
  40. Levicnik-Stezinar S., Rahne-Potokar U., Candotti D., Lelie N., Allain J. P..( 2008;). Anti-HBs positive occult hepatitis B virus carrier blood infectious in two transfusion recipients. . J Hepatol 48: 1022–1025. [CrossRef] [PubMed]
    [Google Scholar]
  41. Lukhwareni A., Burnett R. J., Selabe S. G., Mzileni M. O., Mphahlele M. J..( 2009;). Increased detection of HBV DNA in HBsAg-positive and HBsAg-negative South African HIV/AIDS patients enrolling for highly active antiretroviral therapy at a tertiary hospital. . J Med Virol 412: 406–412. [CrossRef] [PubMed]
    [Google Scholar]
  42. Martin C. M., Welge J. A., Rouster S. D., Shata M. T., Sherman K. E., Blackard J. T..( 2012;). Mutations associated with occult hepatitis B virus infection result in decreased surface antigen expression in vitro. . J Viral Hepat 19: 716–723. [CrossRef] [PubMed]
    [Google Scholar]
  43. Martin C. M., Welge J. A., Shire N. J., Rouster S. D., Shata M. T., Sherman K. E., Blackard J. T..( 2010;). Genomic variability associated with the presence of occult hepatitis B virus in HIV co-infected individuals. . J Viral Hepat 17: 588–597. [CrossRef] [PubMed]
    [Google Scholar]
  44. Martin C. M., Welge J. A., Shire N. J., Shata M. T., Sherman K. E., Blackard J. T..( 2009;). Cytokine expression during chronic versus occult hepatitis B virus infection in HIV co-infected individuals. . Cytokine 47: 194–198. Elsevier Ltd. [CrossRef] [PubMed]
    [Google Scholar]
  45. Mphahlele M. J., Lukhwareni A., Burnett R. J., Moropeng L. M., Ngobeni J. M..( 2006;). High risk of occult hepatitis B virus infection in HIV-positive patients from South Africa. . J Clin Virol 35: 14–20. [CrossRef] [PubMed]
    [Google Scholar]
  46. Ngui S. L., O'Connell S., Eglin R. P., Heptonstall J., Teo C. G..( 1997;). Low detection rate and maternal provenance of hepatitis B virus S gene mutants in cases of failed postnatal immunoprophylaxis in England and Wales. . J Infect Dis 176: 1360–1365. [CrossRef] [PubMed]
    [Google Scholar]
  47. Oon C. J., Tan K. L., Harrison T., Zuckerman A..( 1996;). Natural history of hepatitis B surface antigen mutants in children. . Lancet (London, England) 348: 1524–1525. [CrossRef] [PubMed]
    [Google Scholar]
  48. Pivert A., Servant-Delmas A., Lunel-Fabiani F., Le Guillou-Guillemette H., Laperche S., Ducancelle A..( 2015;). Correlation between the promoter basal core and precore mutations and HBsAg quantification in french blood donors infected with hepatitis B virus. . J Med Virol 87: 529–535. [CrossRef] [PubMed]
    [Google Scholar]
  49. Powell E. A., Gededzha M. P., Rentz M., Rakgole N. J., Selabe S. G., Seleise T. A., Mphahlele M. J., Blackard J. T..( 2014;). Mutations associated with occult hepatitis B in HIV-positive South Africans. . J Med Virol, 1–13.
    [Google Scholar]
  50. Said Z. N..( 2011;). An overview of occult hepatitis B virus infection. . World J Gastroenterol 17: 1927–1938. [CrossRef] [PubMed]
    [Google Scholar]
  51. Seddigh-Tonekaboni S., Lim W. L., Young B., Hou J. L., Waters J., Luo K. X., Thomas H. C., Karayiannis P..( 2001;). Hepatitis B surface antigen variants in vaccinees, blood donors and an interferon-treated patient. . J Viral Hepat 8: 154–158. [CrossRef] [PubMed]
    [Google Scholar]
  52. Shire N. J., Rouster S. D., Rajicic N., Sherman K. E..( 2004;). Occult hepatitis B in HIV-infected patients. . J Acquir Immune Defic Syndr 36: 869–875. [CrossRef] [PubMed]
    [Google Scholar]
  53. Svicher V., Cento V., Bernassola M., Neumann-Fraune M., Van Hemert F., Chen M., Salpini R., Liu C., Longo R. et al.( 2012;). Novel HBsAg markers tightly correlate with occult HBV infection and strongly affect HBsAg detection. . Antiviral Res 93: 86–93. [CrossRef] [PubMed]
    [Google Scholar]
  54. Tseng T. C., Liu C. J., Yang H. C., Chen C. L., Yang W. T., Tsai C. S., Kuo S. F., Verbree F. C., Su T. H. et al.( 2015;). Higher proportion of viral basal core promoter mutant increases the risk of liver cirrhosis in hepatitis B carriers. . Gut 64: 292–302. [CrossRef] [PubMed]
    [Google Scholar]
  55. Tu W.-H..( 2015;). Precore/basal core promoter mutants quantification throughout phases of hepatitis B virus infection by simpleprobe. . World J Gastroenterol 21: 6639.[CrossRef]
    [Google Scholar]
  56. Wiseman E., Fraser M. A., Holden S., Glass A., Kidson B. L., Heron L. G., Maley M. W., Ayres A., Locarnini S. A., Levy M. T..( 2009;). Perinatal transmission of hepatitis B virus: An Australian experience. . Med J Aust 190: 489–492.[PubMed]
    [Google Scholar]
  57. Wu C., Zhang X., Tian Y., Song J., Yang D., Roggendorf M., Lu M., Chen X..( 2010;). Biological significance of amino acid substitutions in hepatitis B surface antigen (HBsAg) for glycosylation, secretion, antigenicity and immunogenicity of HBsAg and hepatitis B virus replication. . J Gen Virol 91: 483–492. [CrossRef] [PubMed]
    [Google Scholar]
  58. Yoshida E. M., Ramji A., Erb S. R., Davis J. E., Steinbrecher U. P., Sherlock C. H., Scudamore C. H., Chung S. W., Williams M., Gutfreund K. S..( 2000;). De novo acute hepatitis B infection in a previously vaccinated liver transplant recipient due to a strain of HBV with a met 133 thr mutation in the "a" determinant. . Liver 20: 411–414. [CrossRef] [PubMed]
    [Google Scholar]
  59. Yuen M. F., Wong D. K., Lee C. K., Tanaka Y., Allain J. P., Fung J., Leung J., Lin C. K., Sugiyama M. et al.( 2011;). Transmissibility of hepatitis B virus (HBV) infection through blood transfusion from blood donors with occult HBV infection. . Clin Infect Dis 52: 624–632. [CrossRef] [PubMed]
    [Google Scholar]
  60. Zuckerman J. N., Zuckerman A. J..( 2003;). Mutations of the surface protein of hepatitis B virus. . Antiviral Res 60: 75–78. [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.000469
Loading
/content/journal/jgv/10.1099/jgv.0.000469
Loading

Data & Media loading...

Supplementary File 2



PDF

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error