NUCLEOTIDE SUBSTITUTIONS IN HEPATITIS B VIRUSES DERIVED FROM CHRONIC HBV PATIENTS

Main Article Content

Narjes Shokatpour
Maryam Vaezjalali
Graham R Foster
Shahnaz Sali

Keywords

Abstract

Background: Mutations in the S gene (HBsAg), precore (PC) and basic core promoter (BCP) of the hepatitis B virus (HBV) are correlated with a wide spectrum of diseases. This study assessed the frequency of mutations in the S gene, PC and BCP regions in chronic hepatitis B (CHB) patients.


 


Methods: 104 CHB patients who visited Tehran Hepatitis centers, were included. The viral load of samples was determined based on the TaqMan method. Regions of the S gene, PC and BCP were amplified by the nested PCR. Positive PCR products were sequenced and analyzed.


 


Results: Successfully sequenced S gene region revealed all the derived strains were genotype D, with the majority (90%) belonging to the ayw2 subtype, and the rest (9%) to the ayw1 subtype. The prevalence of mutations was found to be 51% and 18% in the HBsAg and MHR regions, respectively. 70% of amino acid changes within HBsAg occurred in different immune epitopes, of which 27% and 72% were located in B cell and Th epitopes, respectively. Successfully sequenced PC and BCP regions showed at least one mutation in 84.6% of the patients. The PC and BCP mutations were G1896A (61%), G1899A (23%), A1762T/G1764A (23%) and G1764T/C1766G (26%). None of the strains with A1762T/G1764A mutation carried the G1764T/C1766G mutant.


 


Conclusions: Our results showed common mutations within HBsAg, occurring in immune epitopes, a high rate of G1896A mutations in the PC region, and a negative correlation between the emergence of A1762T/G1764A mutation and the G1764T/C1766G mutant in the BCP region.


 

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References

Agarwal A, Sen S, Banerjee D, Srivastava R, Praharaj A: Distribution of
hepatitis B virus genotype and cancer predicting precore and basal core
promoter mutations. Medical Journal Armed Forces India 2015,
71(3):225-232.
https://doi.org/10.1016/j.mjafi.2015.04.003
PMid:26288490 PMCid:PMC4534539
2. Wang X-L, Ren J-P, Wang X-Q, Wang X-H, Yang S-F, Xiong Y:
Mutations in pre-core and basic core promoter regions of hepatitis B
virus in chronic hepatitis B patients. World Journal of Gastroenterology
2016, 22(11):3268.
https://doi.org/10.3748/wjg.v22.i11.3268
PMid:27004005 PMCid:PMC4790003
3. Assih M, Ouattara AK, Diarra B, Yonli AT, Compaore TR, ObiriYeboah D, Djigma FW, Karou S, Simpore JJWjoh: Genetic diversity of
hepatitis viruses in West-African countries from 1996 to 2018. 2018,
10(11):807.
https://doi.org/10.4254/wjh.v10.i11.807
PMid:30533182 PMCid:PMC6280160
4. Echevarría JM, Avellón A: Hepatitis B virus genetic diversity. Journal
of medical virology 2006, 78(S1):S36-S42.
https://doi.org/10.1002/jmv.20605
PMid:16622876
5. Pujol FH, Navas M-C, Hainaut P, Chemin I: Worldwide genetic
diversity of HBV genotypes and risk of hepatocellular carcinoma.
Cancer letters 2009, 286(1):80-88.
https://doi.org/10.1016/j.canlet.2009.07.013
PMid:19683385
6. Pourkarim MR, Sharifi Z, Soleimani A, Amini?Bavil?Olyaee S,
Elsadek Fakhr A, Sijmons S, Vercauteren J, Karimi G, Lemey P, Maes
P: Evolutionary analysis of HBV "S" antigen genetic diversity in Iranian
blood donors: a nationwide study. Journal of medical virology 2014,
86(1):144-155.
https://doi.org/10.1002/jmv.23798
PMid:24150816
7. Khedive A, Norouzi M, Ramezani F, Karimzadeh H, Alavian S,
Malekzadeh R, Montazeri G, Nejatizadeh A, Ziaee M, Abedi F:
Hepatitis B virus surface protein mutations clustered mainly in CTL
immune epitopes in chronic carriers: results of an Iranian nationwide
study. Journal of viral hepatitis 2013, 20(7):494-501.
https://doi.org/10.1111/jvh.12045
PMid:23730843
8. Petit M-A, Maillard P, Capel F, Pillot J: Immunochemical structure of
the hepatitis B surface antigen vaccine-II. Analysis of antibody
responses in human sera against the envelope proteins. Molecular
immunology 1986, 23(5):511-523.
https://doi.org/10.1016/0161-5890(86)90114-8
9. Ramezani F, Norouzi M, Sarizade GR, Poortahmasebi V, Kalantar E,
Magnius L, Norder H, Domingo E, Jazayeri SM: Mutation hot spots in
hepatitis B surface antigen in chronic carriers from Khoozestan province,
southern of Iran. Iranian Journal of Allergy, Asthma and Immunology
2013, 12(3):269.
Caligiuri P, Cerruti R, Icardi G, Bruzzone B: Overview of hepatitis B virus mutations and their implications in the management of infection. World Journal of Gastroenterology 2016, 22(1):145.
https://doi.org/10.3748/wjg.v22.i1.145
PMid:26755866 PMCid:PMC4698481
11. Hunt CM, McGill JM, Allen MI, Condreay LD: Clinical relevance of hepatitis B viral mutations. Hepatology 2000, 31(5):1037-1044.
https://doi.org/10.1053/he.2000.6709
PMid:10796877
12. Kramvis A, Kew M: The core promoter of hepatitis B virus. Journal of viral hepatitis 1999, 6(6):415-427.
https://doi.org/10.1046/j.1365-2893.1999.00189.x
PMid:10607259
13. Bozday? AM, Bozkaya H, Türky?lmaz AR, Sarýodlu M, Çetinkaya H, Karayalç?n S, Yurdayd?n C, Uzunalimo?lu ÖJJocv: Nucleotide divergences in the core promoter and precore region of genotype D hepatitis B virus in patients with persistently elevated or normal ALT levels. 2001, 21(1):91-101.
https://doi.org/10.1016/S1386-6532(01)00148-2
14. Jazayeri M, Basuni A, Sran N, Gish R, Cooksley G, Locarnini S, Carman WJJovh: HBV core sequence: definition of genotype?specific variability and correlation with geographical origin. 2004, 11(6):488-501.
https://doi.org/10.1111/j.1365-2893.2004.00534.x
PMid:15500549
15. Mohebbi S, Amini?Bavil?Olyaee S, Zali N, Noorinayer B, Derakhshan F, Chiani M, Nejad MR, Antikchi M, Sabahi F, Zali M: Molecular epidemiology of hepatitis B virus in Iran. Clinical microbiology and infection 2008, 14(9):858-866.
https://doi.org/10.1111/j.1469-0691.2008.02053.x
PMid:18844687
16. Ma Q, Wang Y: Comprehensive analysis of the prevalence of hepatitis B virus escape mutations in the major hydrophilic region of surface antigen. Journal of medical virology 2012, 84(2):198-206.
https://doi.org/10.1002/jmv.23183
PMid:22170538
17. Echevarría JM, Avellón A: Improved detection of natural hepatitis B virus surface antigen (HBsAg) mutants by a new version of the VITROS® HBsAg assay. Journal of medical virology 2008, 80(4):598-602.
https://doi.org/10.1002/jmv.21146
PMid:18297712
18. Diarra B, Yonli AT, Sorgho PA, Compaore TR, Ouattara AK, Zongo WA, Tao I, Traore L, Soubeiga ST, Djigma FWJMjoh et al: Occult hepatitis B virus infection and associated genotypes among HBsAg-negative subjects in Burkina Faso. Mediterr J Hematol Infect Dis. 2018 1;10(1):e2018007. doi: 10.4084/MJHID.2018.007. eCollection 2018.
https://doi.org/10.4084/mjhid.2018.007
PMid:29326804 PMCid:PMC5760064
19. Moradi A, Zhand S, Ghaemi A, Javid N, Tabarraei A: Mutations in the S gene region of hepatitis B virus genotype D in Golestan Province-Iran. Virus Genes 2012, 44(3):382-387.
https://doi.org/10.1007/s11262-012-0715-z
PMid:22274739
20. Norouzi M, Ghorashi SA, Ataei B, Yaran M, Malekzadeh R, Alavian SM, Judaki MA, Ghamari S, Namazi A, Rahimnia R: Hepatitis B virus surface antigen variants clustered within immune epitopes in chronic hepatitis B carriers from Hormozgan Province, south of Iran. Iranian journal of basic medical sciences 2010, 13(4):213-224.
21. Nordin M, Ingman M, Lindqvist B, Kidd?Ljunggren K: Variability in the precore and core promoter region of the hepatitis B virus genome. Journal of medical virology 2014, 86(3):437-445.
https://doi.org/10.1002/jmv.23839
PMid:24249691
22. Constantinescu I, Dinu A-A, Boscaiu V, Niculescu M: Hepatitis B Virus Core Promoter Mutations in Patients With Chronic Hepatitis B and Hepatocellular Carcinoma in Bucharest, Romania. Hepatitis monthly 2014, 14(10).
https://doi.org/10.5812/hepatmon.22072
PMid:25477976 PMCid:PMC4250966
23. Ducancelle A, Pivert A, Bertrais S, Boursier J, Balan V, Veillon P, Guillou?Guillemette H, Thibault V, Castelain S, Roquebert B:
Different precore/core mutations of hepatitis B interact with, limit or favor liver fibrosis severity. Journal of gastroenterology and hepatology 2016.
https://doi.org/10.1111/jgh.13338
PMid:26992056
24. Yoo BC, Park J-W, Kim HJ, Lee DH, Cha YJ, Park SM: Precore and core promoter mutations of hepatitis B virus and hepatitis B e antigen-negative chronic hepatitis B in Korea. Journal of hepatology 2003, 38(1):98-103.
https://doi.org/10.1016/S0168-8278(02)00349-5
25. Ghabeshi S, Sharifi Z, Hosseini SM, Shooshtari MM: Correlation between viral load of HBV in chronic hepatitis B patients and precore and Basal core promoter mutations. Hepatitis monthly 2013, 13(2).
https://doi.org/10.5812/hepatmon.7415
PMid:23599717 PMCid:PMC3628088
26. Moradi A, Zhand S, Ghaemi A, Javid N, Bazouri M, Tabarraei A: Mutations in pre-core and basal-core promoter regions of hepatitis B virus in chronic HBV patients from Golestan, Iran. Iranian journal of basic medical sciences 2014, 17(5):370.
27. Fariba Soleimani, Seyed Ali Mohammmad Arabzadeh, Hamidreza Mollaie, ZahraI ranmanesh, Najmeh Nikpour, Motahar M: Evaluation of the frequency of precore/core mutation in patients with chronic hepatitis B, Kerman, Southeast of Iran. Asian pacific journal of tropical disease 2016, 6(8):603-607.
https://doi.org/10.1016/S2222-1808(16)61093-9
28. Tillmann H, Trautwein C, Walker D, Michitaka K, Kubicka S, Böker K, Manns M: Clinical relevance of mutations in the precore genome of the hepatitis B virus. Gut 1995, 37(4):568-573.
https://doi.org/10.1136/gut.37.4.568
PMid:7489947 PMCid:PMC1382912
29. Chan HL, Leung NW, Hussain M, Wong ML, Lok AS: Hepatitis B e antigen-negative chronic hepatitis B in Hong Kong. Hepatology 2000, 31(3):763-768.
https://doi.org/10.1002/hep.510310330
PMid:10706570
30. Fang ZL, Ling R, Wang SS, Nong J, Huang CS, Harrison TJ: HBV core promoter mutations prevail in patients with hepatocellular carcinoma from Guangxi, China. Journal of medical virology 1998, 56(1):18-24.
https://doi.org/10.1002/(SICI)1096-9071(199809)56:1<18::AID-JMV4>3.0.CO;2-Q
31. Poustchi H, Mohamadkhani A, Bowden S, Montazeri G, Ayres A, Revill P, Farrell G, Locarnini S, George J, Malekzadeh R: Clinical significance of precore and core promoter mutations in genotype D hepatitis B?related chronic liver disease. Journal of viral hepatitis 2008, 15(10):753-760.
https://doi.org/10.1111/j.1365-2893.2008.00998.x
PMid:18507754
32. Fujiwara K, Tanaka Y, Orito E, Ohno T, Kato T, Sugihara K, Hasegawa I, Sakurai M, Ito K, Ozasa A: Distribution of HBV genotypes among HBV carriers in Benin: phylogenetic analysis and virological characteristics of HBV genotype E. World journal of gastroenterology: WJG 2005, 11(41):6410-6415.
https://doi.org/10.3748/wjg.v11.i41.6410
PMid:16425408 PMCid:PMC4355778
33. Suppiah J, Zain RM, Bahari N, Nawi SH, Saat Z: G1896A Precore Mutation and Association With HBeAg Status, Genotype and Clinical Status in Patients With Chronic Hepatitis B. Hepatitis monthly 2015, 15(10).
https://doi.org/10.5812/hepatmon.31490
PMid:26587040 PMCid:PMC4644636
34. Baha W, Ennaji MM, Lazar F, Melloul M, El Fahime E, El Malki A, Bennani A: HBV genotypes prevalence, precore and basal core mutants in Morocco. Infection, Genetics and Evolution 2012, 12(6):1157-1162.
https://doi.org/10.1016/j.meegid.2012.04.026
PMid:22579480
35. Sendi H, Mehrab-Mohseni M, Zali MR, Norder H, Magnius LO: T1764G1766 core promoter double mutants are restricted to Hepatitis B virus strains with an A1757 and are common in genotype D. Journal of general virology 2005, 86(9):2451-2458.
https://doi.org/10.1099/vir.0.81023-0
PMid:16099903