Regulatory B cells (Bregs) are reported to play an important role in the immune responses to chronic hepatitis B virus (HBV) via Toll-like-receptors (TLRs) signaling. We aimed to investigate the characteristics of Bregs and the expressions of TLRs in Bregs in the peripheral blood of different immune phases of patients with chronic HBV infection. In this study, we determined the frequencies of Bregs and TLR9, TLR2, and TLR4 in peripheral blood using flow cytometry and determined the levels of IL-10 in serum with the Human Magnetic Cytokine/Chemokine Bead Panel. The results showed that the frequencies of CD19(+) B cells and Bregs were elevated in patients with chronic HBV infection compared with healthy control (HC). The frequencies of Bregs were significantly increased in the immune active group compared with the immune tolerant group and HC group. Meanwhile, the frequencies of Bregs were higher in inactive carrier state group compared with HC group. The levels of serum IL-10 were elevated in the immune active group compared with immune tolerant, inactive carrier state and HC groups. Serum IL-10 levels were positively correlated with alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the immune active group. Compared with HC group, the expression of TLR9 in Bregs reduced in chronic hepatitis B (CHB) patients. However, the expression of TLR2 in Bregs increased in CHB patients. There were no differences of the TLRs expressions in Bregs among various immune phases of chronic HBV infection. These data suggest that the frequencies of Bregs and levels of serum IL-10 were different in various immune phases in patients with chronic HBV infection. Bregs may play some important role in modulating the immune responses of chronic HBV infection. However, whether Bregs modulate immune responses via TLR signaling in chronic HBV infection remains ambiguous.

Objective To investigate the frequencies of regulatory B cells (Bregs) in the peripheral blood and the levels of serum interleukin-10 (IL-10) and interferon-γ-induced protein 10 (IP-10) in different immune phases of chronic hepatitis B virus (HBV) infection. Methods Total of sixty-three patients with chronic HBV infection, while 16 healthy controls were enrolled. The frequencies of Bregs (CD24hiCD38hi B cells) in the peripheral blood were measured by lfow cytometry. IL-10 and IP-10 levels were determined with the Human Magnetic Cytokine/Chemokine Bead Panel on the MAGPIX instrument. Results Compared to immune tolerant phase (IT) and healthy controls (HC), the frequencies of Bregs were signiifcantly elevated in the immune reactive phase (IA) [ (7.89 ± 3.37)%vs (4.77 ± 2.42%), F=9.27, P=0.010;(7.89 ± 3.37)%vs (3.83 ± 2.14)%, F=16.55, P<0.001 ]. The frequencies of mature B cells (CD24intCD38int B cells) were signiifcantly, P=0.02]. Serum IL-10 and IP-10 levels were also elevated signiifcantly in IA group as compared with IT, inactive HBV carrier state (IC) and HC group [(22.53 ± 24.81) pg/ml vs (0.69 ± 1.34) pg/ml, (22.53 ± 24.81) pg/ml vs (0.31 ± 1.12) pg/ml, (22.53 ± 24.81) pg/ml vs (0.003 ± 0.009) pg/ml, P all < 0.001;(2 540.19 ± 1 870.73) pg/ml vs (720.52 ± 285.73) pg/ml, (2 540.19 ± 1 870.73) pg/ml vs (567.38 ± 208.72) pg/ml, (2 540.19 ± 1 870.73) pg/ml vs (624.80 ± 274.45) pg/ml, P all<0.001]. The frequencies of Bregs positively correlated with IL-10 and ALT levels (r = 0.282, P = 0.025; r = 0.305, P = 0.026) in chronic HBV infection. In addition, IL-10 and IP-10 levels were also positively correlated with ALT levels (r = 0.715, P < 0.001; r = 0.653, P < 0.001) in immune reactive phase (IA). Conclusions The frequencies of Bregs, IL-10 and IP-10 levels were elevated significantly in immune reactive phase (IA) of chronic HBV infection. The frequencies of Bregs positively correlated with IL-10 and ALT levels in chronic HBV infection. IL-10 and IP-10 levels were also positively correlated with ALT levels in immune reactive phase (IA) of chronic HBV infection.

AIM:To investigate the association of serum gamma-glutamyl transferase (GGT) levels with chronic hepatitis B infection and hepatitis B e antigen (HBeAg) seroconversion.METHODS:A retrospective study was performed on clinical data collected from patients who had been positive for hepatitis B surface antigen for > 6 mo and who were antiviral-treatment naïve (n = 215) attending the Hepatitis Clinic at Nanjing Drum Tower Hospital between August 2010 and December 2013. Healthy individuals without liver disease (n = 83) were included as controls. Patients were categorized into four groups based on disease status as recommended by the European Association for the Study of the Liver: immune tolerance (IT; n = 47), HBeAg-positive hepatitis (EPH; n = 93), HBeAg-negative hepatitis (ENH; n = 20), and inactive carrier (IC; n = 55). Prediction of complete response (CR) based on serum GGT was also examined in EPH patients (n = 33) treated for 48 wk with nucleos(t)ide analogue (NA) therapy, including lamivudine plus adefovir combination therapy (n = 20) or entecavir monotherapy (n = 13). CR was defined as a serum hepatitis B virus DNA level < 500 copies/mL and HBeAg seroconversion by 48 wk of treatment.RESULTS:Serum GGT levels were significantly increased in EPH and ENH patients relative to the IT, IC, and healthy control groups (P < 0.01 for all). However, no significant difference in serum GGT levels was found between the EPH and ENH groups. Baseline serum GGT levels were significantly higher in patients who achieved CR (7/33; 21.2%) compared to patients in the non-CR group (26/33; 78.8%; P = 0.011). In addition, the decline in serum GGT was greater in CR patients compared to non-CR patients after 24 wk and 48 wk of treatment (P = 0.012 and P = 0.008, respectively). The receiver operating characteristic curve yielded a sensitivity of 85.71% and a specificity of 61.54% at a threshold value of 0.89 times the upper limit of normal for baseline serum GGT in the prediction of CR following NA therapy.CONCLUSION:Serum GGT is significantly elevated in EPH and ENH patients and is a potential biomarker for the prediction of HBeAg seroconversion following NA therapy.

AIM:To investigate the association between red cell distribution width (RDW) and the severity of hepatitis B virus (HBV)-related liver diseases.METHODS:Sixty-nine patients with chronic hepatitis B (CHB) and 61 patients with HBV-related liver cirrhosis were enrolled in the present study. Forty-one healthy individuals were included as controls. Hematological parameters, hepatitis B e-antigen (HBeAg) status, HBV DNA levels and liver biochemistry were analyzed. Child-Pugh scores and Model for End-Stage Liver Disease (MELD) scores of the patients with HBV-related liver cirrhosis were calculated.RESULTS:The RDW was significantly higher in patients with HBV-related liver cirrhosis as compared with CHB patients and healthy controls. RDW was slightly higher in CHB patients as compared with healthy controls. An increasing correlation of RDW with Child-Pugh grades was found. RDW was positively correlated with Child-Pugh scores and MELD scores. In patients with HBV-related liver cirrhosis, RDW was also positively correlated with total bilirubin and negatively correlated with hemoglobin and serum albumin concentration. However, no significant difference was found between HBeAg positive and negative patients and no significant correlation between RDW and HBV DNA levels was found.CONCLUSION:The RDW was elevated in CHB patients and patients with HBV-related liver cirrhosis and was positively correlated with the severity of HBV-related liver cirrhosis. RDW is a potential index to assess the severity of HBV-related liver diseases.