PD-1 blockade partially recovers dysfunctional virus–specific B cells in chronic hepatitis B infection
Loghman Salimzadeh, … , Patrick T.F. Kennedy, Antonio Bertoletti
Loghman Salimzadeh, … , Patrick T.F. Kennedy, Antonio Bertoletti
Published August 7, 2018
Citation Information: J Clin Invest. 2018;128(10):4573-4587. https://doi.org/10.1172/JCI121957.
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Research Article Hepatology Infectious disease

PD-1 blockade partially recovers dysfunctional virus–specific B cells in chronic hepatitis B infection

Abstract

Chronic HBV (CHB) infection suppresses virus-specific T cells, but its impact on humoral immunity has been poorly analyzed. Here, we developed a dual-staining method that utilizes hepatitis B virus (HBV) surface antigens (HBsAg) labeled with fluorochromes as “baits” for specific ex vivo detection of HBsAg-specific B cells and analysis of their quantity, function, and phenotype. We studied healthy vaccinated subjects (n = 18) and patients with resolved (n = 21), acute (n = 11), or chronic (n = 96) HBV infection and observed that frequencies of circulating HBsAg-specific B cells were independent of HBV infection status. In contrast, the presence of serum HBsAg affected function and phenotype of HBsAg-specific B cells that were unable to mature in vitro into Ab-secreting cells and displayed an increased expression of markers linked to hyperactivation (CD21lo) and exhaustion (PD-1). Importantly, B cell alterations were not limited to HBsAg-specific B cells, but affected the global B cell population. HBsAg-specific B cell maturation could be partially restored by a method involving the combination of the cytokines IL-2 and IL-21 and CD40L-expressing feeder cells and was further boosted by the addition of anti–PD-1 Abs. In conclusion, HBV infection has a marked impact on global and HBV-specific humoral immunity, yet HBsAg-specific B cells are amenable to a partial rescue by B cell–maturing cytokines and PD-1 blockade.

Authors

Loghman Salimzadeh, Nina Le Bert, Charles-A. Dutertre, Upkar S. Gill, Evan W. Newell, Christian Frey, Magdeleine Hung, Nikolai Novikov, Simon Fletcher, Patrick T.F. Kennedy, Antonio Bertoletti

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Figure 5

Functional characterization of HBsAg-specific B cells during acute hepatitis B.

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Functional characterization of HBsAg-specific B cells during acute hepat...
(A) HBsAg-specific B cells were sorted from PBMCs of 3 patients at different time points after onset of acute hepatitis B (AHB). The schematic graph on the top indicates the different serological and clinical parameters (ALT, HBsAg, and HBV DNA) at which PBMCs were collected. Sorted HBsAg-specific B cells were polyclonal stimulated with CpG, sCD40L, IL-2, IL-10, and IL-15 for 4 days and subsequently cultured with IL-2, IL-6, IL-10, and IL-15 for another 3 days. After 7 days, culture supernatants were collected and tested in an anti-HBs–specific ELISA. Bars indicate the optical density of detected anti-HBs Ab. (B) HBsAg-specific B cells were sorted from PBMCs of 2 additional acute hepatitis B patients at the indicated time points. Sorted HBsAg-specific B cells were expanded on CD40L-expressing fibroblasts with the addition of IL-2 and IL-21 for 13 days. Expanded cells were tested on anti-HBs B cell ELISpot. Bars indicate the numbers of spots obtained.