Substitutions of rtL228 and/or L229 are involved in the regulation of replication and HBsAg secretion in hepatitis B virus, and do not affect susceptibility to nucleos(t)ide analogs

Qin,Bo; Zhou,Yechao; Zhou,Guozhong; Xu,Xiuping; Wang,Yanan; Chen,Jinkun

doi:10.3892/mmr.2017.7778

Substitutions of rtL228 and/or L229 are involved in the regulation of replication and HBsAg secretion in hepatitis B virus, and do not affect susceptibility to nucleos(t)ide analogs

Authors:
- Bo Qin
- Yechao Zhou
- Guozhong Zhou
- Xiuping Xu
- Yanan Wang
- Jinkun Chen
View Affiliations

Affiliations: Department of Clinical Laboratory Center, Shaoxing People's Hospital, Shaoxing Hospital of Zhejiang University, Shaoxing, Zhejiang 312000, P.R. China, Shaoxing Center for Disease Control and Prevention, Shaoxing, Zhejiang 312000, P.R. China
Published online on: October 12, 2017 https://doi.org/10.3892/mmr.2017.7778
Pages: 9678-9684

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Nucleos(t)ide analogs (NAs) are widely used in the treatment of hepatitis B virus (HBV) and human immunodeficiency virus (HIV). The mutation L210W of HIV‑1 reverse transcriptase (RT) is one of the six principal mutations which confer in vivo resistance to zidovudine. Due to the similar 3D‑structure and high conservation between HIV‑RT and HBV‑RT, the present study aimed to clarify whether corresponding mutations in HBV may decrease its susceptibility to relevant NAs. Mutations including rtL228C/W, rtL229W and rtL228W/L229W were introduced into a HBV replication competent plasmid by fusion polymerase chain reaction. Replication capacity, HBs/e antigen (Ag) levels and susceptibility to NAs were subsequently analyzed in vitro. Single or combination mutations of rtL228 and rtL229 impaired HBV replication. Decreased HBsAg secretion in the supernatant and production in the cell lysate wasobserved with single rtL229W or in combination with rtL228W, while there was no significant difference between wild‑type and mutant HBV with regard to the level of HBeAg in the supernatant and susceptibility to commonly‑used NAs. Substitution mutations of rtL228 and/or L229 in HBV did not alter the susceptibility of the virus to NAs, although replication and HBsAg secretion were affected.

View Figures

View References

1	Li W and Urban S: Entry of hepatitis B and hepatitis D virus into hepatocytes: Basic insights and clinical implications. J Hepatol. 64(1 Suppl): S32–S40. 2016. View Article : Google Scholar : PubMed/NCBI
2	Zhang Q, Liao Y, Chen J, Cai B, Su Z, Ying B, Lu X, Tao C and Wang L: Epidemiology study of HBV genotypes and antiviral drug resistance in multi-ethnic regions from Western China. Sci Rep. 5:174132015. View Article : Google Scholar : PubMed/NCBI
3	Bartholomeusz A, Tehan BG and Chalmers DK: Comparisons of the HBV and HIV polymerase and antiviral resistance mutations. Antivir Ther. 9:149–160. 2004.PubMed/NCBI
4	Dandri M, Lutgehetmann M and Petersen J: Experimental models and therapeutic approaches for HBV. Semin Immunopathol. 35:7–21. 2013. View Article : Google Scholar : PubMed/NCBI
5	Zoulim F and Durantel D: Antiviral therapies and prospects for a cure of chronic hepatitis B. Cold Spring Harb Perspect Med. 5:a0215012015. View Article : Google Scholar : PubMed/NCBI
6	Zoulim F and Locarnini S: Hepatitis B virus resistance to nucleos(t)ide analogues. Gastroenterology. 137:1593–1608.e1-2. 2009. View Article : Google Scholar : PubMed/NCBI
7	Cabuang LM, Shaw T, Littlejohn M, Colledge D, Sozzi V, Soppe S, Warner N, Thompson A, Preiss S, Lam N, et al: In vitro replication phenotype of a novel (−1G) hepatitis B virus variant associated with HIV co-infection. J Med Virol. 84:1166–1176. 2012. View Article : Google Scholar : PubMed/NCBI
8	Petersen J, Thompson AJ and Levrero M: Aiming for cure in HBV and HDV infection. J Hepatol. 65:835–848. 2016. View Article : Google Scholar : PubMed/NCBI
9	Qin B, Budeus B, Cao L, Wu C, Wang Y, Zhang X, Rayner S, Hoffmann D, Lu M and Chen X: The amino acid substitutions rtP177G and rtF249A in the reverse transcriptase domain of hepatitis B virus polymerase reduce the susceptibility to tenofovir. Antiviral Res. 97:93–100. 2013. View Article : Google Scholar : PubMed/NCBI
10	Aragri M, Alteri C, Battisti A, Di Carlo D, Minichini C, Sagnelli C, Bellocchi MC, Pisaturo MA, Starace M, Armenia D, et al: Multiple Hepatitis B virus (HBV) quasispecies and immune-escape mutations are present in HBV surface antigen and reverse transcriptase of patients with acute hepatitis B. J Infect Dis. 213:1897–1905. 2016. View Article : Google Scholar : PubMed/NCBI
11	Caviglia GP, Abate ML, Noviello D, et al: Hepatitis B core-related antigen kinetics in chronic HBV-genotype-D infected patients treated with nucleos(t)ide analogues or pegylated-interferon-alpha. Hepatol Res. 2016.
12	Kim HS, Yim HJ, Jang MK, Park JW, Suh SJ, Seo YS, Kim JH, Kim BH, Park SJ, Lee SH, et al: Management of entecavir-resistant chronic hepatitis B with adefovir-based combination therapies. World J Gastroenterol. 21:10874–10882. 2015. View Article : Google Scholar : PubMed/NCBI
13	Qin B, Zhang B, Zhang X, He T, Xu W, Fu L and Tu C: Substitution rtq267h of hepatitis B virus increases the weight of replication and lamivudine resistance. Hepat Mon. 13:e121602013. View Article : Google Scholar : PubMed/NCBI
14	Yahi N, Tamalet C, Tourres C, Tivoli N and Fantini J: Mutation L210W of HIV-1 reverse transcriptase in patients receiving combination therapy. Incidence, association with other mutations and effects on the structure of mutated reverse transcriptase. J Biomed Sci. 7:507–513. 2000. View Article : Google Scholar : PubMed/NCBI
15	Clavel F and Hance AJ: HIV drug resistance. N Engl J Med. 350:1023–1035. 2004. View Article : Google Scholar : PubMed/NCBI
16	Delviks-Frankenberry KA, Lengruber RB, Santos AF, Silveira JM, Soares MA, Kearney MF, Maldarelli F and Pathak VK: Connection subdomain mutations in HIV-1 subtype-C treatment-experienced patients enhance NRTI and NNRTI drug resistance. Virology. 435:433–441. 2013. View Article : Google Scholar : PubMed/NCBI
17	Qin B, He T, Chen Z, Xu W, Pan G and Tu C: A novel method for the analysis of drug-resistant phenotypes of hepatitis B virus. Int J Mol Med. 31:975–981. 2013. View Article : Google Scholar : PubMed/NCBI
18	Pokrovsky AG, Pronayeva TR, Fedyuk NV, Shirokova EA, Khandazhinskaya AL, Tarusova NB, Karpenko IL and Krayevsky AA: Anti-HIV activity of novel phosphonate derivatives of AZT, d4T and ddA. Nucleosides, Nucleotides Nucleic Acids. 20:767–769. 2001. View Article : Google Scholar : PubMed/NCBI
19	Su M, Xiang K, Li Y, Li Y, Deng J, Xu X, Yan L, Zhuang H and Li T: Higher detection rates of amino acid substitutions in HBV reverse transcriptase/surface protein overlapping sequence is correlated with lower serum HBV DNA and HBsAg levels in HBeAg-positive chronic hepatitis B patients with subgenotype B2. Infect Genet Evol. 40:275–281. 2016. View Article : Google Scholar : PubMed/NCBI
20	Khan N, Guarnieri M, Ahn SH, Li J, Zhou Y, Bang G, Kim KH, Wands JR and Tong S: Modulation of hepatitis B virus secretion by naturally occurring mutations in the S gene. J Virol. 78:3262–3270. 2004. View Article : Google Scholar : PubMed/NCBI
21	Abou-Jaoudé G, Molina S, Maurel P and Sureau C: Myristoylation signal transfer from the large to the middle or the small HBV envelope protein leads to a loss of HDV particles infectivity. Virology. 365:204–209. 2007. View Article : Google Scholar : PubMed/NCBI
22	Li MW, Hou W, Wo JE and Liu KZ: Character of HBV (hepatitis B virus) polymerase gene rtM204V/I and rtL180M mutation in patients with lamivudine resistance. J Zhejiang Univ Sci B. 6:664–667. 2005. View Article : Google Scholar : PubMed/NCBI
23	Lucifora J, Xia Y, Reisinger F, Zhang K, Stadler D, Cheng X, Sprinzl MF, Koppensteiner H, Makowska Z, Volz T, et al: Specific and nonhepatotoxic degradation of nuclear hepatitis B virus cccDNA. Science. 343:1221–1228. 2014. View Article : Google Scholar : PubMed/NCBI
24	Fourati S, Malet I, Guenzel CA, Soulie C, Maidou-Peindara P, Morand-Joubert L, Wirden M, Sayon S, Peytavin G, Simon A, et al: E17A mutation in HIV-1 Vpr confers resistance to didanosine in association with thymidine analog mutations. Antiviral Res. 93:167–174. 2012. View Article : Google Scholar : PubMed/NCBI
25	Michel M, Lone YC, Centlivre M, Roux P, Wain-Hobson S and Sala M: Optimisation of secretion of recombinant HBsAg virus-like particles: Impact on the development of HIV-1/HBV bivalent vaccines. Vaccine. 25:1901–1911. 2007. View Article : Google Scholar : PubMed/NCBI
26	Lopez-Terrada D, Cheung SW, Finegold MJ and Knowles BB: Hep G2 is a hepatoblastoma-derived cell line. Hum Pathol. 40:1512–1515. 2009. View Article : Google Scholar : PubMed/NCBI
27	Pei R, Qin B, Zhang X, Zhu W, Kemper T, Ma Z, Trippler M, Schlaak J, Chen X and Lu M: Interferon-induced proteins with tetratricopeptide repeats 1 and 2 are cellular factors that limit hepatitis B virus replication. J Innate Immun. 6:182–191. 2014. View Article : Google Scholar : PubMed/NCBI