Association of characteristics of HBV quasispecies with hepatitis B surface antigen seroconversion after pegylated interferon-α-2a treatment in child patients

Abstract

Background

The correlation between hepatitis B surface antigen (HBsAg) seroconversion and the characteristics of HBV quasispecies (QS) before and during pegylated interferon-α-2a (PEG-IFN-α-2a) treatment in hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) children has not yet been reported.

Methods

35 patients, including 18 HBsAg seroconverters (SS) and 17 non-seroconverters (SN), were enrolled. Serum samples were collected before treatment and at weeks 12 and 24 of treatment. Sequences within the basal core promoter/pre-core (BCP/PC) and S/reverse transcriptase (S/RT) region were analysed by next-generation sequencing.

Results

There was no significant difference in the baseline diversity of HBV QS (Shannon entropy [Sn]; Hamming distance [HD]) in either region between the two groups. The baseline mutations A1762T/G1764A, C1913A, and T2003A/G or C2004T were correlated with non-response to therapy (P=0.025, P=0.036, P=0.032, respectively). After 24 weeks of therapy, HBV diversity within the BCP/PC region in the SS group notably declined (Sn: P=0.002; HD: P=0.011), while that of the SN group was nearly unchanged. As for the S/RT region, 24 weeks of treatment made no significant difference on QS diversity in either group.

Conclusions

Our data demonstrated that the baseline viral mutations and dynamic changes in HBV QS diversity within the BCP/PC region were closely related to HBsAg seroconversion in HBeAg-positive CHB children treated with PEG-IFN-α-2a.

References

Luo

, Li

, Ruan

Impact of the implementation of a vaccination strategy on hepatitis B virus infections in China over a 20-year period.

Int J Infect Dis 2012; 16: e82–e88.

Chen

H.L.

, Lin

L.H.

, Hu

F.C.

Effects of maternal screening and universal immunization to prevent mother-to-infant transmission of HBV.

Gastroenterology 2012; 142: 773–781.

Chang

M.H.

, You

S.L.

, Chen

C.J.

Decreased incidence of hepatocellular carcinoma in hepatitis B vaccinees: a 20-year follow-up study.

J Natl Cancer Inst 2009; 101: 1348–1355.

Chen

C.H.

, Chen

Y.Y.

, Chen

G.H.

Hepatitis B virus transmission and hepatocarcinogenesis: a 9 year retrospective cohort of 13676 relatives with hepatocellular carcinoma.

J Hepatol 2004; 40: 653–659.

Chang

M.H.

Prevention of hepatocellular carcinoma by universal vaccination against hepatitis B virus: the effect and problems.

Clin Cancer Res 2005; 11: 7953–7957.

Thio

C.L.

, Guo

, Xie

, Nelson

K.E.

, Ehrhardt

Global elimination of mother-to-child transmission of hepatitis B: revisiting the current strategy.

Lancet Infect Dis 2015; 15: 981–985.

Sokal

E.M.

, Paganelli

, Wirth

Management of chronic hepatitis B in childhood: ESPGHAN clinical practice guidelines: consensus of an expert panel on behalf of the European Society of Pediatric Gastroenterology, Hepatology and Nutrition.

J Hepatol 2013; 59: 814–829.

Sokal

E.M.

, Conjeevaram

H.S.

, Roberts

E.A.

Interferon alfa therapy for chronic hepatitis B in children: a multinational randomized controlled trial.

Gastroenterology 1998; 114: 988–995.

Lau

G.K.

, Piratvisuth

, Luo

K.X.

Peginterferon alfa-2a, lamivudine, and the combination for HBeAg-positive chronic hepatitis B.

N Engl J Med 2005; 352: 2682–2695.

10.

Janssen

H.L.

, van Zonneveld

, Senturk

Pegylated interferon alfa-2b alone or in combination with lamivudine for HBeAg-positive chronic hepatitis B: a randomised trial.

Lancet 2005; 365: 123–129.

11.

Buster

E.H.

, Flink

H.J.

, Cakaloglu

Sustained HBeAg and HBsAg loss after long-term follow-up of HBeAg-positive patients treated with peginterferon alpha-2b.

Gastroenterology 2008; 135: 459–467.

12.

Wirth

, Zhang

, Hardikar

Efficacy and safety of peginterferon alfa-2a (40KD) in children with chronic hepatitis B: The PEG-B-ACTIVE Study.

Hepatology 2018; doi: 10.1002/hep.30050.

13.

Domingo

, Sheldon

, Perales

Viral quasispecies evolution.

Microbiol Mol Biol Rev 2012; 76: 159–216.

14.

Chotiyaputta

, Lok

A.S.

Hepatitis B virus variants.

Nat Rev Gastroenterol Hepatol 2009; 6: 453–462.

15.

Lim

S.G.

, Cheng

, Guindon

Viral quasi-species evolution during hepatitis Be antigen seroconversion.

Gastroenterology 2007; 133: 951–958.

16.

Bayliss

, Yuen

, Rosenberg

Deep sequencing shows that HBV basal core promoter and precore variants reduce the likelihood of HBsAg loss following tenofovir disoproxil fumarate therapy in HBeAg-positive chronic hepatitis B.

Gut 2017; 66: 2013–2023.

17.

Sonneveld

M.J.

, Rijckborst

, Zeuzem

Presence of precore and core promoter mutants limits the probability of response to peginterferon in hepatitis B e antigen-positive chronic hepatitis B.

Hepatology 2012; 56: 67–75.

18.

Yang

H.C.

, Chen

C.L.

, Shen

Y.C.

Distinct evolution and predictive value of hepatitis B virus precore and basal core promoter mutations in interferon-induced hepatitis B e antigen seroconversion.

Hepatology 2013; 57: 934–943.

19.

Salpini

, Colagrossi

, Bellocchi

M.C.

Hepatitis B surface antigen genetic elements critical for immune escape correlate with hepatitis B virus reactivation upon immunosuppression.

Hepatology 2015; 61: 823–833.

20.

Charuworn

, Hengen

P.N.

, Aguilar

S.R.

Baseline interpatient hepatitis B viral diversity differentiates HBsAg outcomes in patients treated with tenofovir disoproxil fumarate.

J Hepatol 2015; 62: 1033–1039.

21.

Bruder Costa

, Dufeu-Duchesne

, Leroy

Pegylated interferon α-2a triggers NK-cell functionality and specific T-cell responses in patients with chronic HBV infection without HBsAg seroconversion.

PLoS One 2016; 11: e0158297.

22.

Liu

Y.Z.

, Hou

F.Q.

, Ding

Pegylated interferon alpha enhances recovery of memory T cells in e antigen positive chronic hepatitis B patients.

Virol J 2012; 9: 274.

23.

Domingo

, Martin

, Perales

Viruses as quasispecies: biological implications.

Curr Top Microbiol Immunol 2006; 299: 51–82.

24.

Lin

Y.Y.

, Liu

, Chien

W.H.

New insights into the evolutionary rate of hepatitis B virus at different biological scales.

J Virol 2015; 89: 3512–3522.

25.

Beitz

TEXshade: shading and labeling of multiple sequence alignments using LATEX2 epsilon.

Bioinformatics 2000; 16: 135–139.

26.

Tamura

, Stecher

, Peterson

, Filipski

, Kumar

MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0.

Mol Biol Evol 2013; 30: 2725–2729.

27.

Charuworn

, Hengen

P.N.

, Aguilar

S.R.

Baseline interpatient hepatitis B viral diversity differentiates HBsAg outcomes in patients treated with tenofovir disoproxil fumarate.

J Hepatol 2015; 62: 1033–1039.

28.

Yang

, Liu

, Yang

Quasispecies characteristics in mother-to-child transmission of hepatitis B virus by next-generation sequencing.

J Infect 2017; 75: 48–58.

29.

Le Pogam

, Yuan

T.T.

, Sahu

G.K.

, Chatterjee

, Shih

Low-level secretion of human hepatitis B virus virions caused by two independent, naturally occurring mutations (P5T and L60V) in the capsid protein.

J Virol 2000; 74: 9099–9105.

30.

Lee

, Kim

, Lee

S.A.

Upregulation of endoplasmic reticulum stress and reactive oxygen species by naturally occurring mutations in hepatitis B virus core antigen.

J Gen Virol 2015; 96: 1850–1854.

31.

Zhang

, Ren

, Wu

The L60V variation in hepatitis B virus core protein elicits new epitope-specific cytotoxic T lymphocytes and enhances viral replication.

J Virol 2013; 87: 8075–8084.

32.

Mizokami

, Orito

, Ohba

Constrained evolution with respect to gene overlap of hepatitis B virus.

J Mol Evol 1997; 44 Suppl 2: S83–S90.