Immunological biomarkers as indicators for outcome after discontinuation of nucleos(t)ide analogue therapy in patients with HBeAg negative chronic hepatitis B

The optimal duration of treatment with nucleos(t)ide analogues (NAs) for patients with HBeAg‐negative Chronic Hepatitis B (CHB) is unknown. The aim of this study was to identify an immune signature associated with off‐treatment remission to NA therapy. We performed microarray analysis of PBMCs from six patients with chronic hepatitis B who stopped NA therapy (3 with off‐treatment remission, 3 with relapse) and 5 patients with chronic HBV infection (previously termed “inactive carriers”) served as controls. Results were validated using qRT‐PCR on a second group of 21 individuals (17 patients who stopped treatment and 4 controls). PBMCs from 38 patients on long‐term NA treatment were analysed for potential to stop treatment. Microarray analysis indicated that patients with off‐treatment remission segregated as a distinct out‐group. Twenty‐one genes were selected for subsequent validation. Ten of these were expressed at significantly lower levels in the patients with off‐treatment remission compared to the patients with relapse and predicted remission with AUC of 0.78‐0.92. IFNγ, IL‐8, FASLG and CCL4 were the most significant by logistic regression. Twelve (31.6%) of 38 patients on long‐term NA therapy had expression levels of all these four genes below cut‐off values, and hence were candidates for stopping treatment. Our data suggest that patients with HBeAg‐negative CHB who remain in off‐treatment remission 3 years after NA cessation have a distinct immune signature and that PBMC RNA levels of IFNγ, IL‐8, FASLG and CCL4 may serve as potential biomarkers for stopping NA therapy.