Frequency of drug resistance and immune escape mutations in the hepatitis B virus genome detected in pregnant women in the Republic of Guinea

Cover Page

Cite item

Full Text

Abstract

The aim of the work is to assess the prevalence of hepatitis B virus drug resistance mutations and immune escape mutations in pregnant women in the Republic of Guinea.

Materials and methods. Blood plasma samples obtained from 480 pregnant women from different regions of the Republic of Guinea with laboratory-confirmed viral hepatitis B were studied. Nucleotide sequences for genotype identification and mutation detection were obtained using nested-PCR followed by Sanger sequencing, based on overlapping pairs of primers spanning the complete genome of the virus.

Results and discussion. In the examined group, the viral genotype E was the most prevalent (92.92%) compared with subgenotypes A1 (1.67%), A3 (1.46%), D1 (0.63%), D2 (1.04%) and D3 (2.29%). Among the examined HBV-infected pregnant women, 188 (39.17%) had undetectable HBsAg. Drug resistance mutations were detected in 33 individuals, which amounted to 6.88%. The following mutations were found: S78T (27.27%), L80I (24.24%), S202I (15.15%), M204I/V (42.42%). The presence of polymorphic variants not described as drug resistant has also been shown in positions associated with the development of drug resistance to tenofovir, lamivudine, telbivudine and entecavir (L80F, S202I, M204R). When analyzing the MHR and the region of a determinant, mutations were detected in 318 (66.25%) of pregnant women. In 172 of them, which amounted to 54.09%, multiple mutations were found. The amino acid substitutions in 13 positions associated with HBsAg-negative hepatitis B and/or potentially affecting HBsAg antigenicity were identified.

Conclusion. The high prevalence of immune escape and drug resistance mutations potentially associated with false-negative result of HBsAg screening, prophylaxis failure, and virological failure of therapy that has been identified among treatment naive pregnant women imposes a serious problem.

About the authors

T.A.L. Balde

Research Institute of Applied Biology of Guinea

Email: thiernoamadoulabe.balde@gmail.com
ORCID iD: 0000-0002-3808-4380

Researcher

Guinea, Kindia

Yulia V. Ostankova

Saint-Petersburg Pasteur Institute

Author for correspondence.
Email: shenna1@yandex.ru
ORCID iD: 0000-0003-2270-8897

PhD, Senior Researcher at the Laboratory of Molecular Immunology, Head of the Laboratory of Immunology and Virology HIV Infection

Russian Federation, St. Petersburg

S. Boumbaly

Research Institute of Applied Biology of Guinea; Centre International de Recherche sur les Infections Tropicales en Guinée

Email: drboumbaly@yahoo.fr
ORCID iD: 0000-0002-4506-6033

PhD, chief graduate school

Guinea, Kindia; N’Zérékoré

Ekaterina V. Naidenova

Russian Research Anti-Plague Institute “Microbe”

Email: katim2003@mail.ru
ORCID iD: 0000-0001-6474-3696

PhD, Senior Researcher, Laboratory of Virology, Department of Infectious Diseases Diagnostics

Russian Federation, Saratov

Elena B. Zueva

Saint-Petersburg Pasteur Institute

Email: ezueva75@mail.ru
ORCID iD: 0000-0002-0579-110X

PhD, Biologist at the Department of Diagnostics of HIV Infection and AIDS-Associated Diseases

Russian Federation, St. Petersburg

Elena N. Serikova

Saint-Petersburg Pasteur Institute

Email: elena.donetsk.serikova@mail.ru
ORCID iD: 0000-0002-0547-3945

Researcher, Laboratory of Immunology and Virology HIV Infection, Postgraduate Student

Russian Federation, St. Petersburg

Diana E. Valutite

Saint-Petersburg Pasteur Institute

Email: dianavalutite008@gmail.com
ORCID iD: 0000-0002-0931-102X

clinical laboratory diagnostics doctor at at the Department of Diagnostics of HIV Infection and AIDS-Associated Diseases Infection

Russian Federation, St. Petersburg

Alexander N. Schemelev

Saint-Petersburg Pasteur Institute

Email: tvildorm@gmail.com
ORCID iD: 0000-0002-3139-3674

Junior Researcher, Laboratory of Immunology and Virology HIV Infection, Postgraduate Student

Russian Federation, St. Petersburg

Vladimir S. Davydenko

Saint-Petersburg Pasteur Institute

Email: vladimir_david@mail.ru
ORCID iD: 0000-0003-0078-9681

Junior Researcher, Laboratory of Immunology and Virology HIV Infection, Postgraduate Student

Russian Federation, St. Petersburg

Elena V. Esaulenko

Saint-Petersburg Pasteur Institute

Email: eve-gpmu@mail.ru
ORCID iD: 0000-0003-3669-1993

PhD, MD (Medicine), Professor, Head at the Department of Infectious Diseases of Adults and Epidemiology, Head at the laboratory of viral hepatitis

Russian Federation, St. Petersburg

Areg A. Totolian

Saint-Petersburg Pasteur Institute

Email: otolian@pasteurorg.ru
ORCID iD: 0000-0003-4571-8799

Academician of the Russian Academy of Sciences, PhD, MD (Medicine), Professor, Head at the Laboratory of Molecular Immunology, Director; head Department of Immunology

Russian Federation, St. Petersburg

References

  1. WHO. Hepatitis B. Key facts. Available at: http://www.who.int/news-room/fact-sheets/detail/hepatitis-b
  2. WHO. Global progress report on HIV, viral hepatitis and sexually transmitted infections, 2021: Accountability for the global health sector strategies 2016–2021: Actions for impact. Web Annex 2: Data Methods. Available at: https://www.who.int/publications/i/item/9789240027077
  3. GBD 2019 Hepatitis B Collaborators. Global, regional, and national burden of hepatitis B, 1990–2019: a systematic analysis for the Global Burden of Disease Study 2019. Lancet Gastroenterol. Hepatol. 2022; 7(9): 796–829. https://doi.org/10.1016/S2468-1253(22)00124-8
  4. Schweitzer A., Horn J., Mikolajczyk R.T., Krause G., Ott J.J. Estimations of worldwide prevalence of chronic hepatitis B virus infection: a systematic review of data published between 1965 and 2013. Lancet. 2015; 386(10003): 1546–55. https://doi.org/10.1016/S0140-6736(15)61412-X
  5. Kao J.H. Hepatitis B vaccination and prevention of hepatocellular carcinoma. Best Pract. Res. Clin. Gastroenterol. 2015; 29(6): 907–17. https://doi.org/10.1016/j.bpg.2015.09.011
  6. Yao N., Fu S., Wu Y., Tian Z., Feng Y., Li J., et al. Incidence of mother-to-child transmission of hepatitis b in relation to maternal peripartum antiviral prophylaxis: A systematic review and meta-analysis. Acta Obstet. Gynecol. Scand. 2022; 101(11): 1197–206. https://doi.org/10.1111/aogs.14448
  7. Pan C.Q., Duan Z., Dai E., Zhang S., Han G., Wang Y., et al. Tenofovir to prevent hepatitis B transmission in mothers with high viral load. N. Engl. J. Med. 2016; 374(24): 2324–34. https://doi.org/10.1056/NEJMoa1508660
  8. Terrault N.A., Lok A.S.F., McMahon B.J., Chang K.M., Hwang J.P., Jonas M.M., et al. Update on prevention, diagnosis, and treatment of chronic hepatitis B: aASLD 2018 hepatitis B guidance. Hepatology. 2018; 67(4): 1560–99. https://doi.org/10.1002/hep.29800
  9. Nassal M. HBV cccDNA: viral persistence reservoir and key obstacle for a cure of chronic hepatitis B. Gut. 2015; 64(12): 1972–84. https://doi.org/10.1136/gutjnl-2015-309809
  10. Akinbodewa A.A., Gbadegesin B.A., Adejumo O.A., Ahmed S.D., Uwameiye O., Dada S.A., et al. A multicentre study of awareness and practice of vaccination against infectious diseases among haemo-dialysis subjects in Nigeria. West Afr. J. Med. 2019; 36(3): 239–45.
  11. Kramvis A. Molecular characteristics and clinical relevance of African genotypes and subgenotypes of hepatitis B virus. S. Afr. Med. J. 2018; 108(8b): 17–21. https://doi.org/10.7196/SAMJ.2018.v108i8b.13495
  12. Stasi C., Silvestri C., Voller F. Emerging trends in epidemiology of hepatitis B virus infection. J. Clin. Transl. Hepatol. 2017; 5(3): 272–6. https://doi.org/10.14218/JCTH.2017.00010
  13. Mokaya J., McNaughton A.L., Hadley M.J., Beloukas A., Geretti A.M., Geodhals D. A systematic review of hepatitis B virus (HBV) drug and vaccine escape mutations in Africa: A call for urgent action. PLoS Negl. Trop. Dis. 2018; 12(8): e0006629. https://doi.org/10.1371/journal.pntd.0006629
  14. Phan N.M.H., Faddy H.M., Flower R.L., Dimech W.J., Spann K.M., Roulis E.V. Low genetic diversity of hepatitis B virus surface gene amongst Australian blood donors. Viruses. 2021; 13(7): 1275. https://doi.org/10.3390/v13071275
  15. Hossain M.G., Ueda K. Investigation of a novel hepatitis B virus surface antigen (HBsAg) escape mutant affecting immunogenicity. PLoS One. 2017; 12(1): e0167871. https://doi.org/10.1371/journal.pone.0167871
  16. Ostankova Yu.V., Serikova E.N., Semenov A.V., Totolyan Areg A. Method for hepatitis B virus DNA detecting in biological material at low viral load based on nested PCR with detection on three viral targets in real-time mode. Klinicheskaya laboratornaya diagnostika. 2022; 67(9): 530–7. https://doi.org/10.51620/0869-2084-2022-67-9-530-537 https://elibrary.ru/qndbzr (in Russian)
  17. Popova A.Yu., Kutyreva V.V., Totolyan Areg A., eds. Hepatitis B in West Africa: Epidemiology, Diagnosis, Prevention [Gepatit B v stranakh Zapadnoy Afriki: epidemiologiya, diagnostika, profilaktika]. St. Petersburg; 2021. (in Russian)
  18. Garmiri P., Loua A., Haba N., Candotti D., Allain J.P. Deletions and recombinations in the core region of hepatitis B virus genotype E strains from asymptomatic blood donors in Guinea, west Africa. J. Gen. Virol. 2009; 90(Pt. 10): 2442–51. https://doi.org/10.1099/vir.0.012013-0
  19. Balde T.A.L., Bumbali S., Serikova E.N., Valutite D.E., Shchemelev A.N., Ostankova Yu.V., et al. Comparative analysis of the vertical risk of transmission of some blood-borne infections in the Republic of Guinea. Problemy osobo opasnykh infektsiy. 2021; (1): 87–94. https://doi.org/10.21055/0370-1069-2021-1-87-94 https://elibrary.ru/upnyfx (in Russian)
  20. Boumbaly S., Serikova E.N., Balde T.A.L., Ostankova Yu.V., Schemelev A.N., Valutite D.E., et al. Amino acid substitutions in core and HBsAg regions of hepatitis B virus in patients with monoinfection and HBV/HIV-coinfection in the Republic of Guinea. HIV Infection and Immunosuppressive Disorders. 2021; 13(3): 122–33. https://doi.org/10.22328/2077-9828-2021-13-3-122-133
  21. Bumbali S., Balde T.L., Semenov A.V., Ostankova Yu.V., Serikova E.N., Naydenova E.V., et al. Prevalence of viral hepatitis B markers among blood donors in the Republic of Guinea. Voprosy virusologii. 2022; 67(1): 59–68. https://doi.org/10.36233/0507-4088-92 https://elibrary.ru/zybhjz (in Russian)
  22. Ingasia L.A.O., Kostaki E.G., Paraskevis D., Kramvis A. Global and regional dispersal patterns of hepatitis B virus genotype E from and in Africa: A full-genome molecular analysis. PLoS One. 2020; 15(10): e0240375. https://doi.org/10.1371/journal.pone.0240375
  23. Shirvani-Dastgerdi E., Winer B.Y., Celià-Terrassa T., Kang Y., Tabernero D., Yagmur E., et al. Selection of the highly replicative and partially multidrug resistant rtS78T HBV polymerase mutation during TDF-ETV combination therapy. J. Hepatol. 2017; 67(2): 246–54. https://doi.org/10.1016/j.jhep.2017.03.027
  24. Colagrossi L., Hermans L.E., Salpini R., Di Carlo D., Pas S.D., Alvarez M., et al. HEPVIR working group of the European Society for translational antiviral research (ESAR). Immune-escape mutations and stop-codons in HBsAg develop in a large proportion of patients with chronic HBV infection exposed to anti-HBV drugs in Europe. BMC Infect. Dis. 2018; 18(1): 251. https://doi.org/10.1186/s12879-018-3161-2
  25. Sloan R.D., Ijaz S., Moore P.L., Harrison T.J., Teo C.G., Tedder R.S. Antiviral resistance mutations potentiate hepatitis B virus immune evasion through disruption of its surface antigen a determinant. Antivir. Ther. 2008; 13(3): 439–47.
  26. Wang M.L., Tang H. Nucleos(t)ide analogues causes HBV S gene mutations and carcinogenesis. Hepatobiliary Pancreat. Dis. Int. 2016; 15(6): 579–86. https://doi.org/10.1016/s1499-3872(16)60064-4
  27. Lai M.W., Liang K.H., Yeh C.T. Hepatitis B virus preS/S truncation mutant rtM204I/sW196* increases carcinogenesis through deregulated HIF1A, MGST2, and TGFbi. Int. J. Mol. Sci. 2020; 21(17): 6366. https://doi.org/10.3390/ijms21176366
  28. Gencay M., Hübner K., Gohl P., Seffner A., Weizenegger M., Neofytos D., et al. Ultra-deep sequencing reveals high prevalence and broad structural diversity of hepatitis B surface antigen mutations in a global population. PLoS One. 2017; 12(5): e0172101. https://doi.org/10.1371/journal.pone.0172101
  29. Lazarevic I., Banko A., Miljanovic D., Cupic M. Immune-escape hepatitis B virus mutations associated with viral reactivation upon immunosuppression. Viruses. 2019; 11(9): 778. https://doi.org/10.3390/v11090778
  30. Adesina O.A., Akanbi O.A., Opaleye O.O., Japhet M.O., Wang B., Oluyege A.O., et al. Detection of Q129H immune escape mutation in apparently healthy hepatitis B virus carriers in southwestern Nigeria. Viruses. 2021; 13(7): 1273. https://doi.org/10.3390/v13071273
  31. Faleye T.O., Adewumi O.M., Ifeorah I.M., Akere A., Bakarey A.S., Omoruyi E.C., et al. Detection and circulation of hepatitis B virus immune escape mutants among asymptomatic community dwellers in Ibadan, southwestern Nigeria. Int. J. Infect. Dis. 2015; 39: 102–9. https://doi.org/10.1016/j.ijid.2015.08.008
  32. Powell E.A., Gededzha M.P., Rentz M., Rakgole N.J., Selabe S.G., Seleise T.A., et al. Mutations associated with occult hepatitis B in HIV-positive South Africans. J. Med. Virol. 2015; 87(3): 388–400. https://doi.org/10.1002/jmv.24057
  33. Olagbenro M., Anderson M., Gaseitsiwe S., Powell E.A., Gededzha M.P., Selabe S.G., et al. In silico analysis of mutations associated with occult hepatitis B virus (HBV) infection in South Africa. Arch. Virol. 2021; 166(11): 3075–84. https://doi.org/10.1007/s00705-021-05196-7
  34. Urabe A., Imamura M., Tsuge M., Kan H., Fujino H., Fukuhara T., et al. The relationship between HBcrAg and HBV reinfection in HBV related post-liver transplantation patients. J. Gastroenterol. 2017; 52(3): 366–75. https://doi.org/10.1007/s00535-016-1240-y
  35. Rajput M.K. Mutations and methods of analysis of mutations in Hepatitis B virus. AIMS Microbiol. 2020; 6(4): 401–21. https://doi.org/10.3934/microbiol.2020024
  36. Hamada-Tsutsumi S., Iio E., Watanabe T., Murakami S., Isogawa M., Iijima S., et al. Validation of cross-genotype neutralization by hepatitis B virus-specific monoclonal antibodies by in vitro and in vivo infection. PLoS One. 2015; 10(2): e0118062. https://doi.org/10.1371/journal.pone.0118062
  37. World Health Organization (WHO). Hepatitis B vaccination coverage [Internet]. Available online: https://immunizationdata.who.int/pages/coverage/hepb.html access date: 23.06.23
  38. Wilson P., Parr J.B., Jhaveri R., Meshnick S.R. Call to action: prevention of mother-to-child transmission of hepatitis B in Africa. J. Infect. Dis. 2018; 217(8): 1180–3. https://doi.org/10.1093/infdis/jiy028
  39. Bassoum O., Kimura M., Dia A.T., Lemoine M., Shimakawa Y. Coverage and timeliness of birth dose vaccination in sub-Saharan Africa: A systematic review and meta-analysis. Vaccines. 2020; 8(2): 301. https://doi.org/10.3390/vaccines8020301
  40. Moturi E., Tevi-Benissan C., Hagan J., Shendale S., Mayenga D., Murokora D., et al. Implementing a birth dose of hepatitis B vaccine in Africa: Findings from assessments in 5 countries. J. Immunol. Sci. 2018; (Suppl. 5): 31–40. https://doi.org/10.29245/2578-3009/2018/si.1104
  41. Salpini R., Colagrossi L., Bellocchi M.C., Surdo M., Becker C., Alteri C., et al. Hepatitis B surface antigen genetic elements critical for immune escape correlate with hepatitis B virus reactivation upon immunosuppression. Hepatology. 2015; 61(3): 823–33. https://doi.org/10.1002/hep.276

Supplementary files

Supplementary Files
Action
1. JATS XML
Download
2. Fig. 1. Age distribution of pregnant women in the study group.

Download (144KB)
Indexing metadata
3. Fig. 2. Distribution of HBV subgenotypes in the examined group.

Download (40KB)
Indexing metadata
4. Fig. 3. Distribution of HBV serotypes in the examined group.

Download (70KB)
Indexing metadata
5. Fig. 4. Phylogenetic tree for HBV complete genome nucleotide sequences of genotypes A and D isolated from pregnant women living in the Republic of Guinea in comparison with the reference sequences presented in the GenBank database. Reference sequences are designated with GenBank accession numbers and indicated genotype and the region of the sample origin. The Woolly Monkey HBV nucleotide sequence AY226578 was used as the outgroup. The sequences from this study are indicated by white circles (HBsAg+) and black circles (HBsAg–). Bootstrap values ≥ 70.

Download (324KB)
Indexing metadata
6. Fig. 5. Phylogenetic tree for some HBV complete genome nucleotide sequences of genotype Е isolated from pregnant women living in the Republic of Guinea in comparison with the reference sequences presented in the GenBank database. Reference sequences are designated with GenBank accession numbers and indicated genotype and the region of the sample origin. The Woolly Monkey HBV nucleotide sequence AY226578 was used as the outgroup. The samples studied in this work are indicated by white circles (HBsAg+) and black circles (HBsAg–). Bootstrap values ≥ 70.

Download (197KB)
Indexing metadata

Copyright (c) 2023 Balde T., Ostankova Y.V., Boumbaly S., Naidenova E.V., Zueva E.B., Serikova E.N., Valutite D.E., Schemelev A.N., Davydenko V.S., Esaulenko E.V., Totolian A.A.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.

Согласие на обработку персональных данных с помощью сервиса «Яндекс.Метрика»

1. Я (далее – «Пользователь» или «Субъект персональных данных»), осуществляя использование сайта https://journals.rcsi.science/ (далее – «Сайт»), подтверждая свою полную дееспособность даю согласие на обработку персональных данных с использованием средств автоматизации Оператору - федеральному государственному бюджетному учреждению «Российский центр научной информации» (РЦНИ), далее – «Оператор», расположенному по адресу: 119991, г. Москва, Ленинский просп., д.32А, со следующими условиями.

2. Категории обрабатываемых данных: файлы «cookies» (куки-файлы). Файлы «cookie» – это небольшой текстовый файл, который веб-сервер может хранить в браузере Пользователя. Данные файлы веб-сервер загружает на устройство Пользователя при посещении им Сайта. При каждом следующем посещении Пользователем Сайта «cookie» файлы отправляются на Сайт Оператора. Данные файлы позволяют Сайту распознавать устройство Пользователя. Содержимое такого файла может как относиться, так и не относиться к персональным данным, в зависимости от того, содержит ли такой файл персональные данные или содержит обезличенные технические данные.

3. Цель обработки персональных данных: анализ пользовательской активности с помощью сервиса «Яндекс.Метрика».

4. Категории субъектов персональных данных: все Пользователи Сайта, которые дали согласие на обработку файлов «cookie».

5. Способы обработки: сбор, запись, систематизация, накопление, хранение, уточнение (обновление, изменение), извлечение, использование, передача (доступ, предоставление), блокирование, удаление, уничтожение персональных данных.

6. Срок обработки и хранения: до получения от Субъекта персональных данных требования о прекращении обработки/отзыва согласия.

7. Способ отзыва: заявление об отзыве в письменном виде путём его направления на адрес электронной почты Оператора: info@rcsi.science или путем письменного обращения по юридическому адресу: 119991, г. Москва, Ленинский просп., д.32А

8. Субъект персональных данных вправе запретить своему оборудованию прием этих данных или ограничить прием этих данных. При отказе от получения таких данных или при ограничении приема данных некоторые функции Сайта могут работать некорректно. Субъект персональных данных обязуется сам настроить свое оборудование таким способом, чтобы оно обеспечивало адекватный его желаниям режим работы и уровень защиты данных файлов «cookie», Оператор не предоставляет технологических и правовых консультаций на темы подобного характера.

9. Порядок уничтожения персональных данных при достижении цели их обработки или при наступлении иных законных оснований определяется Оператором в соответствии с законодательством Российской Федерации.

10. Я согласен/согласна квалифицировать в качестве своей простой электронной подписи под настоящим Согласием и под Политикой обработки персональных данных выполнение мною следующего действия на сайте: https://journals.rcsi.science/ нажатие мною на интерфейсе с текстом: «Сайт использует сервис «Яндекс.Метрика» (который использует файлы «cookie») на элемент с текстом «Принять и продолжить».