Polyclonal and monoclonal B lymphocytes response in HCV-infected patients treated with direct-acting antiviral agents

Hepatitis C virus (HCV) chronic infection can be associated with extrahepatic manifestations such as mixed cryoglobulinaemia and lymphoproliferative disorders that are endowed with increased rates of morbidity and all-cause mortality. In this study, we used flow cytometry to evaluate the effect of interferon-free antiviral treatment on peripheral blood lymphocytes in HCV-infected patients with or without associated lymphoproliferative disorders. Flow cytometry analysis of peripheral blood lymphocytes was performed at baseline and at the end of treatment. In HCV-infected patients with lymphoproliferative disorders, we evaluated immunoglobulin (Ig) light chain \u3ba/\u3bb ratio variations as a measure of monoclonal B-cell response to antiviral therapy. Healthy volunteers were enrolled as controls. A total of 29 patients were included, nine with and 20 without lymphoproliferative disorders. Sustained virological response was achieved in 29 of 29 patients. We observed a significant reduction in the B-cell compartment (39% global reduction) in eight of nine HCV-infected patients with lymphoproliferative disorders after viral clearance. We recognized the same trend, even if less pronounced, in HCV-infected patients without lymphoproliferative disorders (9% global reduction). Among HCV-infected patients with lymphoproliferative disorders, three showed an improvement/normalization of the immunoglobulin light chain ratio, whereas in the remaining six patients monoclonal B cells persisted to be clonally restricted even 1\ua0year after the end of treatment. Our data show that DAAs treatment can be effective in reducing the frequency of pathological B cells in the peripheral blood of HCV-infected patients affected by HCV-associated lymphoproliferative disorders; however, monoclonal populations can persist after viral eradication

Changes in plasma circulating microvesicles in patients with HCV\u2010related cirrhosis after treatment with direct\u2010acting antivirals

BACKGROUND AND AIMS: The eradication of Hepatitis C (HCV) infection by direct-acting antiviral agents (DAAs) has been linked to an amelioration of liver synthesis and regression of fibrosis. Although changes in number and type of circulating microvesicles (MVs) have been reported in cirrhosis, conclusive data on the effect of DAAs treatment on MVs profile in HCV cirrhotic patients remain scarce. METHODS: We measured levels of endothelial, platelet and hepatocyte MVs, as well as MVs expressing versican core protein (VCAN+) in patients with HCV-related cirrhosis at baseline, end of therapy (EOT), at 12, 24 and 48 weeks (W) after EOT by new generation flow-cytometry. RESULTS: Fifty-eight patients were enrolled (86% Child's A). MVs were increased at EOT versus baseline, though only platelet MVs revealed a statistically significant difference (p< 0.01). MVs levels did not change significantly after EOT notwithstanding a steady downward trend towards baseline levels. Conversely, VCAN+MVs dropped significantly at EOT (p <0.001) and remained low throughout the follow-up. Hepatocyte MVs significantly correlated with liver stiffness (r 0.40, p 0.0021). Eight composite outcomes occurred during the 1-year follow-up: 3 portal vein thromboses, 2 hepatocellular carcinomas and 3 liver decompensation. Child's B, the presence of F2 oesophageal varices (OR for interaction 19.2 [95%CI 1.45-253.7], p 0.023) and platelet MVs (OR 1.026 [95%CI 1.00-1.05, p 0.023) correlated significantly with clinical outcomes. CONCLUSIONS: VCAN+MVs appear to mirror the profibrotic status of the cirrhotic disease; hepatocyte MVs correlate with liver stiffness and increased platelet MVs levels could be associated with a worse clinical outcome. This article is protected by copyright. All rights reserved

Reversal of hypercoagulability in patients with HCV-related cirrhosis after treatment with direct acting antivirals

The long-term impact of sustained virological response (SVR) after direct-acting antivirals (DAAs) on the hypercoagulability associated with HCV cirrhosis is unknown. We longitudinally evaluated the effect of DAAs treatment on cirrhotic coagulopathy. METHODS: Pro- and anticoagulant factors levels and thrombin generation were assessed in patients with HCV-related cirrhosis at baseline, end of therapy (EOT), at 12, 24 and 48 weeks (W) after EOT. RESULTS: Fifty-eight patients were enrolled (86% Child's A). SVR was 100%. Median factor VIII activity significantly decreased at EOT, 12W and 24W compared to baseline, whereas protein C significantly increased at 24W and 48W. Cirrhotic patients showed a slight but sustained increase of endogenous thrombin potential (ETP) with a statistically significant difference at EOT, 12W, 24W and 48W compared to baseline. Conversely, thrombomodulin-modified ETP was elevated before treatment and decreased over time to normal levels at 24W and 48W. The ETP ratio decreased slowly at EOT and 12W, and was significantly decreased at 24W and 48W compared to baseline (p<0.001 for both comparisons), being not statistically different from ETP ratio measured in healthy controls. Child's B patients showed a significantly higher ETP ratio compared to Child's A at baseline and did not show any significant improvement in ETP ratio through 12W. Two Child's B patients developed PVT with an incidence rate of 1.1% p-yrs (95%CI, 0.18 to 3.58). CONCLUSIONS: DAAs therapy in HCV-related cirrhotic patients is associated with significant changes in thrombin generation suggesting a reversal of hypercoagulability particularly in Child's A patients. This article is protected by copyright. All rights reserved

Hepatitis C virus eradication with direct acting antiviral improves insulin resistance

Sustained virological response (SVR) after interferon-based therapy is associated with improvement of insulin resistance (IR) in HCV infected patients. Few data are available in the direct-acting antivirals (DAAs) era, especially in cirrhotic patients. We prospectively evaluated the long-term effect of DAAs on IR. Patients treated with DAAs between May 2015 and December 2016 in 3 tertiary care centers were recruited. Patients with diabetes were excluded. Biochemical and virological data were collected at baseline, 12/24/48-weeks (W) after the end of therapy (EOT). Presence of IR was defined by a "homeostasis model assessment index for IR" [HOMA-IR]) > 2.5. Liver fibroscan was performed at baseline, at 24/48W after EOT. 138 patients were enrolled (mean age 58 years, M/F 85/53, GT1 61%, 68.8% cirrhotic). Sixty-eight (94/138) had IR. Patients with IR had significantly higher stiffness than patients without it (23\ub112 vs. 15\ub18; p <0.0001). SVR12 was achieved in 135 (98%) patients, and 124 (90%) patients reached the 48W post-EOT. At this time-point, the percentage of patients with IR significantly decreased to 49% (p=0,01). HOMA-IR was significantly lower than baseline (1.8 vs. 3; p<0.001), and this was related to a significant reduction of insulin level (11.7\ub16.3 vs 16.4\ub18.3). High BMI was associated with a significantly lower probability of achieving a non-IR status at 24W (p=0.05) and 48W (p=0.03). IN CONCLUSION: SVR following DAAs led to a significant reduction of IR, even in patients with cirrhosis. Nevertheless, IR can persist after the achievement of SVR, especially in patients with high BMI