Распространенность оккультного гепатита B среди HBsAg-негативных лиц с ВИЧ инфекцией в Великом Новгороде.

РЕЗЮМЕ.

Цель. Оценить распространенность оккультного вируса гепатита В среди ВИЧ-инфицированных пациентов с вирусологической неэффективностью антиретровирусной терапии в г. Великий Новгород.

Материалы и методы. В работе были использованы образцы плазмы крови от 76 HBsAg-негативных ВИЧ-инфицированных пациентов с вирусологической неэффективностью антиретровирусной терапии из Великого Новгорода. Для выявления вируса гепатита В выделяли нуклеиновые кислоты с использованием коммерческого набора «АмплиПрайм Рибо-преп». Для амплификации и секвенирующей реакции использовали перекрывающиеся пары специфических праймеров, совместно фланкирующие фрагмент протяженностью 1475 пар оснований (п.о.), включающий рекомендованный для генотипирования вируса гепатита В регион Pre-S1/Pre-S2/S протяженностью 1169 п.о.

Результаты. Среди 76 образцов ДНК вируса гепатита В была обнаружен в 44 образцах, что составило 57,89%. Ни у одного из пациентов не был выявлен HBsAg, при этом у 6 пациентов (13,63%) выявлены антитела HBсor IgG и HBe IgG. На основании филогенетического анализа показано, что в обследованной группе выявлен только генотип D, являющийся наиболее распространенным генотипом вируса гепатита В в Российской Федерации. При этом преобладал субгенотип D2 (47,72%) по сравнению с субгенотипом D1 (34,09%) и субгенотипом D3 (18,18%). Обсуждаются возможные пути распространения нехарактерного для региона вируса гепатита В субгенотипа D1. Выявлен только один изолят вируса гепатита В с мутациями лекарственной устойчивости к терапии нуклеотидных / нуклеозидных аналогов - замещение аминокислот в гене полимеразы вируса (L180M, M204V), связанное с развитием резистентности к ламивудину, энтекавиру, телбивудину и тенофовиру.

Заключение. Высокая распространенность оккультного гепатита В среди ВИЧ-инфицированных пациентов свидетельствует о недостаточности применяемых в настоящее время тестов для диагностики хронических инфекций ВГВ. Идентификация оккультного гепатита В у ВИЧ-инфицированных лиц представляется целесообразной для своевременной терапии пациентов и требует применения более чувствительных методов.

ABSTRACT.

Aim. To estimate the prevalence of the occult hepatitis B virus among HIV-infected patients with the virological ineffectiveness of antiretroviral therapy in Veliky Novgorod.

Materials and methods. Blood plasma samples from 76 HBsAg-negative HIV-infected patients with virological inefficiency of antiretroviral therapy from Veliky Novgorod were used in the work. For the detection of the hepatitis B virus, nucleic acids were isolated using the commercial kit AmplePrime Ribo-prep. For amplification and sequencing, overlapping pairs of specific primers were used, jointly flanking a 1475 base pair (bp) fragment including the 1169 bp Pre-S1 / Pre-S2 / S region recommended for genotyping the hepatitis B virus.

Results. Among 76 samples of hepatitis B virus DNA, 44 samples were found, which was 57,89%. None of the patients had HBsAg, and 6 patients (13,63%) had HBcor IgG and HBe IgG antibodies. On the basis of phylogenetic analysis, it was shown that only genotype D, which is the most common genotype of the hepatitis B virus in the Russian Federation, was detected in the examined group. The subgenotype D2 (47,72%) prevailed in comparison with the subgenotype D1 (34,09%) and the subgenotype D3 (18,18%). The distribution possible ways of the subgenotype D1 hepatitis B virus, which is uncharacteristic for the region, are discussed. Only one isolate of hepatitis B virus with mutations of drug resistance to nucleotide / nucleoside analogue therapy has been identified - amino acid substitution in the polymerase gene of the virus (L180M, M204V) associated with the development of resistance to lamivudine, entecavir, telbivudine and tenofovir.

Conclusion. The high prevalence of occult hepatitis B among HIV-infected patients indicates a lack current tests for the diagnosis of chronic HBV infections. The identification of occult hepatitis B in HIV-infected individuals seems appropriate for timely treatment of patients and requires the use more sensitive methods.

1. Yim H.J., Lok A.S. Natural history of chronic hepatitis B virus infection: what we knew in 1981 and what we know in 2005. Hepatology, 2006, v.43(2 S.1), P. S173–S181.

2. Raimondo G., Allain J. P., Brunetto M. R., Buendia M. A., Chen D. S., Colombo M., Craxi A., Donato F., Ferrari C., Gaeta G. B., Gerlich W. H., Levrero M., Locarnini S., Michalak T., Mondelli M. U., Pawlotsky J. M., Pollicino T., Prati D., Puoti M., Samuel D., Shouval D., Smedile A., Squadrito G., Trepo C., Villa E., Will H., Zanetti A. R., Zoulim F. Statements from the Taormina expert meeting on occult hepatitis B virus infection. J. Hepatol, 2008, 49, P. 652–657.

3. Guo J.T., Guo H. Metabolism and function of hepatitis B virus cccDNA: Implications for the development of cccDNA-targeting antiviral therapeutics. Antiviral research, 2015, vol. 122, P. 91–100.

4. Dandri M., Locarnini S. New insight in the pathobiology of hepatitis B virus infection. Gut, 2012, V. 61, № 1, P.i6–i17.

5. Semenov A.V., Ostankova Yu.V., Faizullaev Kh.N., Kazakova E.I., Kozlov A.V., Musabaev E.I., Totolyan Areg A. HBV covalently closed circular DNA as a marker of prevalence of occult hepatitis В in patients with HBV, HDV and HCV infection in Uzbekistan. Zhurnal mikrobiologii, epidemiologii i immunobiologii = Journal of Microbiology, Epidemiology and Immunobiology, 2016, no. 5, pp. 43–49 (In Russ.).

6. Samal J., Kandpal M., Vivekanandan P. Molecular mechanisms underlying occult hepatitis B virus infection. Clin. Microbiol. Rev., 2012, 25, P. 142–163.

7. Joint United Nations Programme on HIV/AIDS., United Nations. Global Report: UNAIDS Report on the Global AIDS epidemic: 2012. Geneva: UNAIDS; 2012. 103. A5 p. p.

8. WHO Prevention and control of viral hepatitis infection: frame work for global action 2012. Geneva: WHO, 2012.

9. Dadasheva A, E., Mamedov MK, Mikhailov MI About two types of groups with a high risk of infection with hepatitis B and C viruses: epidemiological and clinical significance. In the world of viral hepatitis, 2011, № 1. pp. 12–14 (In Russ.).

10. Nelson P., Mathers B., Cowie B., Hagan H., Jarlais D., Horyniak D., Degenhardt L. The epidemiology of viral hepatitis among people who inject drugs: Results of global systematic reviews. The Lancet, 2011, V. 378, №9791, P. 571–583.

11. Rossi С., Shrier I., Marshall L., Cnossen S., Schwartzman K., Klein M.B., Schwarzer G., Greenaway C. Seroprevalence of chronic hepatitis B virus infection and prior immunity in immigrants and refugees: A systematic review and meta-analysis. PLoS ONE, 2012, V. 7, №9, P. e44611.

12. Argani H., Akhtarishojaie E. Levamizole enhances immune responsiveness to intra-dermal and intra-muscular hepatitis B vaccination in chronic hemodialysis patients. J. Immune Based Ther. Vaccines, 2006, 4, P.3.

13. Alter M.J. Epidemiology of viral hepatitis and HIV co-infection. J. Hepatol., 2006, 44(1 Suppl), P.S6-S9.

14. Burnett R.J., François G., Kew M.C., Leroux-Roels G., Meheus A., Hoosen A.A., et al. Hepatitis B virus and human immunodeficiency virus co-infection in sub-Saharan Africa: a call for further investigation. Liver Int., 2005, 25(2), P.201–213.

15. Kapembwa K.C., Goldman J.D., Lakhi S., Banda Y., Bowa K., Vermund S.H., et al. HIV, Hepatitis B, and Hepatitis C in Zambia. J. Glob. Infect. Dis., 2011, v.3(3), P.269–274.

16. Vallet-Pichard A., Pol S. Natural history and predictors of severity of chronic hepatitis C virus (HCV) and human immunodeficiency virus (HIV) co-infection. J. Hepatol., 2006, 44(1), P.S28–S34.

17. Osborn M.K., Guest J.L., Rimland D. Hepatitis B virus and HIV coinfection: relationship of different serological patterns to survival and liver disease. HIV Med., 2007, 8(5), P.271-279.

18. Demidenko T.P., Neverov V.A. Viral hepatitis. Ed. Yu.V. Lobzin, St. Petersburg, 2011, 229p. (In Russ.).

19. Thio C.L., Seaberg E..C, Skolasky R., et al. HIV-1, hepatitis B virus, and risk of liver-related mortality in the multicenter cohort study (MACS). Lancet, 2002, v.360, p. 1921–1926.

20. Agyeman A.A., Ofori-Asenso R. Prevalence of HIV and hepatitis B coinfection in Ghana: a systematic review and meta-analysis. AIDS. Res. Ther., 2016, vol.13:23.

21. Ramezani A., Banifazl M., Mohraz M., Rasoolinejad M., Aghakhani A. Occult hepatitis B virus infection: A major concern in HIV-infected patients: Occult HBV in HIV. Hepat. Mon., 2011, v.11(1), P.7-10.

22. WHO. Guidelines for the prevention, care and treatment of persons with chronic hepatitis B infection Geneve, Switzerland: WHO, 2015.

23. Mphahlele M.J., Lukhwareni A., Burnett R.J., Moropeng L.M., Ngobeni J.M. High risk of occult hepatitis B virus infection in HIV-positive patients from South Africa. J. Clin. Virol., 2006, v.35(1), P.14–20.

24. Filippini P., Coppola N., Pisapia R., Scolastico C., Marrocco C., Zaccariello A., et al. Impact of occult hepatitis B virus infection in HIV patients naive for antiretroviral therapy. AIDS, 2006, v.20(9), P.1253–1260.

25. Brichler S., Lagathu G., Chekaraou M.A., . Le Gal F, Edouard A., Dény P., Césaire R., Gordien E. African, Amerindian and European hepatitis B virus strains circulate on the Caribbean Island of Martinique. J. Gen. Virol., 2013, vol. 94 (Pt 10), P. 2318-2329.

26. Kumar S., Stecher G., Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution. 2016, vol. 33, №7. P. 1870-1874.

27. Coffin C.S., Mulrooney-Cousins P.M., Osiowy C., van der Meer F., Nishikawa S., Michalak T.I., van Marle G., Gill M.J. Virological characteristics of occult hepatitis B virus in a North American cohort of human immunodeficiency virus type 1-positive patients on dual active anti-HBV/HIV therapy. J. Clin. Virol., 2014, vol.60(4), P.347-353.

28. Gupta S., Singh S. Occult Hepatitis B Virus infection in ART-Naïve HIV-Infected patients seen at a tertiary care center in North India. BMC Infection Diseases, 2010, 10, p.53.

29. Honarmand A., Pourahmad M., Solhjoo K., Kohan M., Davami M.H., Mostafavi Zadeh S.K. Occult hepatitis B demonstrated by anti-HBc and HBV DNA in HIV-positive patients. Rep. Biochem. Mol. Biol., 2014, v.3(1), P.38-42.

30. Alvarez-Munoz M.T., Maldonado-Rodriguez A., Rojas-Montes O., Torres-Ibarra R., Gutierrez-Escolano F., Vazquez-Rosales G., Gomez A., Munoz O., Torres J., Lira R. Occult hepatitis B virus infection among Mexican human immunodeficiency virus-1-infected patients. World J. Gastroenterol., 2014, vol.20(37), P.13530-13537.

31. Bautista-Amorocho H., Castellanos-Domínguez Y.Z., Rodríguez-Villamizar L.A., Velandia-Cruz S.A., Becerra-Peña J.A., Farfán-García A.E. Epidemiology, risk factors and genotypes of HBV in HIV-infected patients in the northeast region of Colombia: high prevalence of occult hepatitis B and F3 subgenotype dominance. PLoS One, 2014, vol.9(12):e114272.

32. Cohen Stuart J.W., Velema M., Schuurman R., Boucher C.A., Hoepelman A.I. Occult hepatitis B in persons infected with HIV is associated with low CD4 counts and resolves during antiretroviral therapy. J. Med. Virol., 2009, vol.81(3), P.441–445.

33. Marite B., Montalvo M.C., Rodriguez Lde L., Sariego S., Verdasquera D., Vincent M., Gutiérrez A., Sánchez M. Occult hepatitis B in Cuban HIV patients. MEDICC Rev., 2011, v.13(2), P.32–37.

34. Fabris P., Biasin M.R., Giordani M.T., Berardo L., Menini V., Carlotto A., et al. Impact of occult HBV infection in HIV/HCV co-infected patients: HBV-DNA detection in liver specimens and in serum samples. Curr. HIV Res., 2008, v.6(2), P.173–179.

35. Esmail M.A., Mahdi W.K., Khairy R.M., Abdalla N.H. Genotyping of occult hepatitis B virus infection in Egyptian hemodialysis patients without hepatitis C virus infection. J. Infect. Public Health, 2016, v.9(4), P.452-457.

36. Kim K.H., Chang H.Y., Park J.Y. Spontaneous HBsAg loss in Korean patients: relevance of viral genotypes, S gene mutations, and covalently closed circular DNA copy numbers. Clin. Mol. Hepatol., 2014, vol. 20 (3), P. 251-260.

37. Ostankova Ju. V., Semenov A.V., Burkitbayev Z.K., Savchuk T.N., Totolian Areg A. Genetic variants of hepatitis B virus in primary donors in Astana, Kazakhstan. Russian Journal of Infection and Immunity. 2016;6(4):359-365. (In Russ.).

38. Wiersma S. T., McMahon B., Pawlotsky J.-M., Thio C. L., Thursz Ma., Gee Lim S., Ocama P., Esmat G., Maimuna M., Bell D., Vitoria M., Eramova I., Lavanchy D., Dusheiko G. WHO. Treatment of chronic hepatitis B virus infection in resource-constrained settings: expert panel consensus. Liver International, 2011, vol.31, Issue 6, p. 755-761.

39. Sagnelli E., Pisaturo M., Martini S., Filippini P., Sagnelli C., Coppola N. Clinical impact of occult hepatitis B virus infection in immunosuppressed patients. World J. Hepatol., 2014, v.6(6), P.384–393.