Circulating and intrahepatic antiviral B cells are defective in hepatitis B
Alice R. Burton, … , Nadege Pelletier, Mala K. Maini
Alice R. Burton, … , Nadege Pelletier, Mala K. Maini
Published August 9, 2018
Citation Information: J Clin Invest. 2018;128(10):4588-4603. https://doi.org/10.1172/JCI121960.
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Research Article Immunology Infectious disease

Circulating and intrahepatic antiviral B cells are defective in hepatitis B

Abstract

B cells are increasingly recognized as playing an important role in the ongoing control of hepatitis B virus (HBV). The development of antibodies against the viral surface antigen (HBV surface antigen [HBsAgs]) constitutes the hallmark of resolution of acute infection and is a therapeutic goal for functional cure of chronic HBV (CHB). We characterized B cells directly ex vivo from the blood and liver of patients with CHB to investigate constraints on their antiviral potential. Unexpectedly, we found that HBsAg-specific B cells persisted in the blood and liver of many patients with CHB and were enriched for T-bet, a signature of antiviral potential in B cells. However, purified, differentiated HBsAg-specific B cells from patients with CHB had defective antibody production, consistent with undetectable anti-HBs antibodies in vivo. HBsAg-specific and global B cells had an accumulation of CD21–CD27– atypical memory B cells (atMBC) with high expression of inhibitory receptors, including PD-1. These atMBC demonstrated altered signaling, homing, differentiation into antibody-producing cells, survival, and antiviral/proinflammatory cytokine production that could be partially rescued by PD-1 blockade. Analysis of B cells within healthy and HBV-infected livers implicated the combination of this tolerogenic niche and HBV infection in driving PD-1hiatMBC and impairing B cell immunity.

Authors

Alice R. Burton, Laura J. Pallett, Laura E. McCoy, Kornelija Suveizdyte, Oliver E. Amin, Leo Swadling, Elena Alberts, Brian R. Davidson, Patrick T.F. Kennedy, Upkar S. Gill, Claudia Mauri, Paul A. Blair, Nadege Pelletier, Mala K. Maini

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Figure 1

B cells specific for HBV surface antigen persist in chronic infection.

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B cells specific for HBV surface antigen persist in chronic infection. (...
(A) Representative staining: HBsAg-specific B cells in a vaccinated HC identified using an AF488–HBsAg bait, compared with FMO. (B) Representative ELISpot well image: anti-HBs–secreting B cells in HBsAg bait sorted– and HBsAg bait–depleted cells from HBV-vaccinated HC (representative of n = 3). anti-HBs measured in supernatant by ELISA (IU/ml). (C) HBsAg-specific B cells (red bars; % of total CD19+CD20+) across the course of HBV vaccination in 2 healthy donors. Samples taken 2 weeks prior to first dose and 7 days after each dose (given 1 and 6 months after the initial dose). Dashed line represents serum anti-HBs titer (IU/ml) determined by ELISA. Red line delineates threshold level of 0.18 based on mean + SD of unexposed controls. (D) Frequency of HBsAg-specific B cells in unexposed HC (n = 24), HBV-HCV+ patients (n = 6), HBV-vaccinated HC (vac HC; n = 29), and patients with CHB (n = 84) identified using AF488–HBsAg bait staining. Red line delineates threshold of detection, as above. (E) Frequency of HBsAg-specific B cells plotted against HBsAg titer (IU/ml; n = 48). (F) Cross-sectional analysis showing the frequency of HBsAg-specific B cells at HBV-acute and HBV-resolved (res.) time points (n = 8). (G) Longitudinal analysis of HBsAg-specific B cells during acute-resolving infection. Frequencies plotted relative to viral load (dashed line; IU/ml), serum ALT (dotted line; IU/liter), and serological status (indicated by bars). (H) anti-HBs in supernatants from stimulated FACS-sorted HBsAg-specific B cells (n = 3 HBV-vaccinated HC; n = 4 patients with CHB). Number of cells ranged from 5 × 103 to 1.2 × 104 for HBV-vaccinated HC and 5 × 103 to 1.7 × 104 in patients with CHB. Representative plot for HBV-vaccinated HC is also shown in Supplemental Figure 1A. Error bars indicate mean ± SEM. P values were determined by Kruskal-Wallis test (ANOVA) with Dunn’s post hoc test for pairwise multiple comparisons (D), Spearman’s rank correlation (E); and Wilcoxon’s paired t test (F). **P < 0.005; ***P < 0.001; ****P < 0.0001.