Circulating serum HBsAg level is a biomarker for HBV-specific T and B cell responses in chronic hepatitis B patients

Chronic hepatitis B (CHB) infection functional cure is defined as sustained loss of HBsAg and several therapeutic strategies are in clinical development designed to pharmacologically reduce serum HBsAg, break immune tolerance, and increase functional cure rates. However, little is known about pre-treatment HBsAg levels as an indicator of HBV immune potential. Here, we compared the phenotypes and HBV-specific response of lymphocytes in CHB patients stratified by serum HBsAg levels <500 (HBs lo ) or >50,000 IU/ml (HBs hi ) using immunological assays (flow cytometry, ICS, ELISPOT). HBs hi patients had significantly higher expression of inhibitory PD-1 on CD4 + T cells, particularly among TEMRA subset, and higher FcRL5 expression on B cells. Upon HBcAg(core) or HBsAg(env)-stimulation, 85% and 60% of HBs lo patients had IFNγ + TNFα + and IFNγ + IL2 + CD4 + T cell responses respectively, in comparison to 33% and 13% of HBs hi patients. Checkpoint blockade with αPD-1 improved HBV-specific CD4 + T cell function only in HBs lo patients. HBsAg-specific antibody-secreting cells (ASCs) response was not different between these groups, yet αPD-1 treatment resulted in significantly higher fold change in ASCs among patients with HBsAg <100 IU/ml compared to patients with HBsAg >5,000 IU/ml. Thus, serum HBsAg correlates with inhibitory receptor expression, HBV-specific CD4 + T cell responses, and augmentation by checkpoint blockade.