Immune-suppression driven Hepatitis B Virus (HBV)-reactivation poses serious concerns since it occurs in several clinical settings and can result in severe forms of hepatitis. Previous studies showed that HBV strains, circulating in patients with HBV-reactivation, are characterized by an enrichment of immune-escape mutations in HBV surface antigen (HBsAg). Here, we focused on specific immune-escape mutations associated with the acquisition of N-linked glycosylation sites in HBsAg (NLGSs). In particular, we investigated profiles of NLGSs in 47 patients with immunosuppression-driven HBV-reactivation and we evaluated their impact on HBsAg-antigenicity and HBV-replication in vitro. At HBV-reactivation, despite a median serum HBV-DNA of 6.7 [5.3-8.0] logIU/mL, 23.4% of patients remained HBsAg-negative. HBsAg-negativity at HBV-reactivation correlated with the presence of >1 additional NLGSs (p < 0.001). These NLGSs are located in the major hydrophilic region of HBsAg (known to be the target of antibodies) and resulted from the single mutation T115N, T117N, T123N, N114ins, and from the triple mutant S113N+T131N+M133T. In vitro, NLGSs strongly alter HBsAg antigenic properties and recognition by antibodies used in assays for HBsAg-quantification without affecting HBsAg-secretion and other parameters of HBV-replication. In conclusion, additional NLGSs correlate with HBsAg-negativity despite HBV-reactivation, and hamper HBsAg-antigenicity in vitro, supporting the role of NGSs in immune-escape and the importance of HBV-DNA for a proper diagnosis of HBV-reactivation.
Summary Here, CD40L expression and cytokine production have been analysed in peripheral blood cells from orthotopic liver transplantation (OLT) recipients treated with ribavirin for recurrent chronic hepatitis C. The study included 18 OLT recipients treated with ribavirin, eight control OLT recipients and 10 healthy controls. FACS analysis showed that baseline expression of CD40L was not different between ribavirin-treated patients and controls. In contrast, after stimulation with both HCV core antigen and phorbol myristate acetate (PMA) plus ionomycin (IO), the expression of CD40L on CD4 lymphocytes was significantly higher in the ribavirin group compared with controls. In the ribavirin group, the increased expression of CD40L significantly correlated with reduction of HCV RNA levels with respect to pretreatment values. Finally, ribavirin treatment was not associated with modification of PMA-IO-induced cytokine production by T lymphocytes and interleukin (IL)-1β and tumour necrosis-α (TNF)-α production by CD40L-stimulated monocytes. In conclusion, these data indicate that ribavirin upmodulates CD40L expression on CD4 T cells, a property which may account in part for its ability to enhance the antiviral activity of interferon-α in the treatment of chronic HCV infection.
The purpose of this study was to compare the spectral characteristics of lipids, choline-containing compounds, and glutamine-glutamate complex assessed with (1)H-MR spectroscopy with the histologic findings in patients with chronic hepatitis C.Nine healthy controls and 30 patients with biopsy-proven hepatitis C virus-related liver disease participated in this prospective study. Degree of fibrosis and histologic activity were scored according to the METAVIR classification. The percentage of involved hepatocytes was used to grade steatosis. Hepatic spectra were obtained with a 3-T spectroscopic system. Tenfold cross-validated stepwise discriminant analysis was performed to classify disease severity on the basis of the spectroscopic findings.There was a strong correlation between (1)H-MR spectroscopically measured lipid concentration and the degree of steatosis at histologic examination (r = 0.9236, p < 0.0001). This finding enabled clear separation of groups according to degree of histologically determined steatosis. Variation in lipid concentration was consistent with the degree of steatosis (r = 0.7265, p < 0.0001) and stage of fibrosis (r = 0.8156, p < 0.0001). In univariate analysis, concentrations of both choline-containing compounds and glutamine-glutamate complex had a direct correlation with histologic grade (p < 0.0001) and degree of steatosis (p < 0.0001) but not with stage of fibrosis (p > 0.05). In multivariate analysis, the only factor independently associated with concentrations of choline-containing compounds and glutamine-glutamate complex was histologic grade. In cross-validated discriminant analysis based on choline-containing compound, glutamine-glutamate complex, and lipid resonance, 70% (21 of 30) of the histologic grade groups and 73% (22 of 30) of the steatosis groups were correctly classified.Hydrogen-1 MR spectroscopy can be an alternative to liver biopsy in the evaluation of steatosis and necroinflammatory activity in liver disease but is not useful for complete evaluation of hepatic fibrosis.
