N.Z. was demonstrated to correlate with B-cell activation and disease progression in patients with human immunodeficiency virus (HIV) infection [29, 31]. Little information is available regarding the characteristics of the CD39/CD73/adenosine pathway in B-cells or its clinical significance in chronic HBV infection. In patients with CHB, we observed skewing of CD39 and CD73 expression on total B-cells, regardless of their specificity. The decreased CD39 and CD73 expression on B-cells was closely associated with viral burden and liver inflammation, and restoration of CD39 and CD73 expression on B-cells was closely associated with antiviral efficacy. Our findings suggested that the CD39/CD73/adenosine pathway has an important role underlying B-cell hyperactivation. Metformin, a clinically available drug, has the potential to regulate B-cell activation, suggesting that intervention in the CD39/CD73/adenosine pathway in B-cells using metformin might represent a therapeutic option for HBV-induced immune pathogenesis in CHB. Materials and methods Patients Two hundred and two patients infected with HBV were enrolled, including 95 treatment-naive patients, who were further categorized into 15 immune tolerance (IT) patients, 45 immune activation (IA) patients, 15 inactive carriers (IC), and 20 immune reactivation (RA) patients, and 107 complete responders (CR), who had received at least 1?year of entecavir treatment and had serum HBV DNA below a detectable level (20?IU/mL), together with alanine aminotransferase (ALT) normalization. In addition, antiviral efficacy was further determined by their HBeAg and HBsAg status [32, 33]. Twenty-five AMG 487 S-enantiomer age-matched healthy controls (HCs) were simultaneously enrolled who tested serologically negative for HBV, Rabbit polyclonal to LOXL1 Hepatitis C virus (HCV), and HIV. The baseline clinical characteristics of these patients and HCs are listed in Table?1. Table 1. Clinical characteristics of enrolled subjects in the study nonparametric test or analysis of variance test was performed between multiple groups. Significant differences between two groups were determined using the MannCWhitney nonparametric test or Students = 15), IA (= 45), IC (= 15), and immune reactivation (RA, = 20). (C) Subjects were categorized into groups determined by their viral load, HBeAg status, HBsAg status, ALT levels, and hepatic necro-inflammation, respectively. CD39- and CD73-expression levels on intra-hepatic B-cells decrease with liver inflammation We further investigated the expression profiles of CD39 and CD73 on B-cells in the livers of CHB patients. As shown in Figure?2, the frequencies of intra-hepatic CD39+, CD73+, and CD39+CD73+ B-cells were decreased further compared with those in peripheral blood, regardless of G or S score grouping. Interestingly, there was a larger reduction in CD39 expression on intra-hepatic B-cells in patients with a higher G score than those with lower G scores. These data indicated that the CD39 and CD73 expression on intra-hepatic B-cells was markedly reduced in the livers of CHB patients and may be associated AMG 487 S-enantiomer with severe liver inflammation. Open in a separate window Figure 2. The expression profiles of CD39 and CD73 on B-cells in the livers of patients with chronic hepatitis B. Thirteen patients were classified by (A) hepatic necro-inflammation grade (G) and (B) fibrotic stage (S), respectively. Frequencies of CD39+, CD73+, and CD39+CD73+ B-cells in peripheral blood mononuclear cells (PBMCs) and liver tissue were determined using flow cytometry. *< 0.05. CD39- and CD73-expression levels on B-cells increase with antiviral efficacy Subsequently, to investigate the expression profiles of CD39 and CD73 on B-cells during antiviral therapy, we performed a cross-sectional study comparing CR patients with different responses to antiviral therapy. As shown in Figure?3A, the frequencies of CD39+, CD73+, and CD39+CD73+ B-cells were increased in HBeAg-negative patients compared with those in HBeAg-positive patients. Further analysis showed that the frequencies of CD39+ and CD39+CD73+ B-cells were increased in patients with lower HBsAg levels, whereas there was no difference in the frequencies of CD73+ B-cells when AMG 487 S-enantiomer the patients were grouped by HBsAg levels (Figure?3B). These data indicated that restoration of CD39 and CD73 expression on B-cells AMG 487 S-enantiomer is closely associated with antiviral efficacy. Open in a separate window Figure 3. CD39 and CD73 expression on.