Hepatic vein outflow obstruction causes congestion of the liver, leading to necrosis, fibrosis, and portal hypertension (PH). Transjugular intrahepatic portosystemic shunt (TIPS) reduces congestion and PH by providing artificial outflow. The aim of the study was to investigate fibrosis progression in patients with Budd-Chiari syndrome (BCS) and TIPS using transient elastography (TE). From 2010 to 2022, 25 patients received 80 TEs using FibroScan® (3.2±2.1 per patient). TIPS function was assessed using Doppler ultrasound or radiological intervention. At the time of TE examination, 21 patients had patent shunts. Four patients had occluded shunts but normal pressure gradients during the intervention. The first TE measurement performed 9.8±6.8 years after the BCS diagnosis showed stiffness values of 24.6±11.5 kPa. A second or last measurement performed 7.0±2.9 years after the first measurement showed similar stiffness values of 24.1±15.7 kPa (p=0.943). Except for three patients, the liver stiffness was always >12 kPa, indicating advanced fibrosis. Stiffness values obtained <5 years (n=8, 23,8±9,2 kPa) or >5 years after the BCS diagnosis (24,9±12,7 kPa) did not differ (p=0.907). In addition, stiffness was not related to the interval between BCS and TIPS implantation (p=0.999). One patient received liver transplantation, and two patients died from non-hepatic causes. Most patients develop mild to moderate cirrhosis, possibly during the early phase of the disease. Timing of TIPS did not influence fibrosis progression. This and the release of portal hypertension may argue in favor of a generous TIPS implantation practice in patients with BCS.

The study of hepatitis C virus (HCV) replication in cell culture is mainly based on cloned viral isolates requiring adaptation for efficient replication in Huh7 hepatoma cells. The analysis of wild type (WT) isolates has been enabled by expression of SEC14L2 and by inhibitors targeting deleterious host factors. Here, we aimed at optimizing cell culture models to allow infection with HCV from patient sera. We used Huh7-Lunet cells ectopically expressing SEC14L2, CD81 and a GFP reporter with nuclear translocation upon cleavage by the HCV protease to study HCV replication, combined with a drug-based regimen for stimulation of non-modified wildtype isolates. RT-qPCR based quantification of HCV infections using patient sera suffered from high background in the daclatasvir treated controls. We therefore established an automated image analysis pipeline based on imaging of whole wells and iterative training of a machine-learning tool, using nuclear GFP localization as a readout for HCV infection. Upon visual validation of hits assigned by the automated image analysis the method revealed no background in daclatasvir treated samples. Thereby, infection events were found for 15 of 34 high titer HCV genotype (gt) 1b sera, revealing a significant correlation of serum titer and successful infection. We further show that transfection of viral RNA extracted from sera can be used in this model as well, albeit with so far limited efficiency. Overall, we generated a robust serum infection assay for gt1b isolates using semi-automated image analysis, which was superior to conventional RT-qPCR based quantification of viral genomes.