Datasheet1_Combination of everolimus and low-dose tacrolimus controls histological liver allograft injury as sufficiently as high-dose tacrolimus.docx

IntroductionThe combination of everolimus (EVR) and low-dose tacrolimus (lowTAC) prevents T cell-mediated rejection of liver grafts as sufficiently as high-dose tacrolimus (highTAC) and mycophenolate, but is associated with a preserved kidney function within the first years after orthotopic liver transplantation (OLT). However, none of the available studies assessed the histological pattern of graft injury or fibrosis in surveillance biopsies (svLbx).MethodsAll svLbx taken under at least one month of stable immunosuppression with either EVR (aim 3-8 ng/ml) combined with lowTAC (aim 3-5 ng/ml) or highTAC (aim 5-8 ng/ml) combined with mycophenolate (500-1500 mg/day) within the first three to four years after OLT at our center were included. Patients who were switched to EVR because of insufficient control of alloreactivity were excluded.ResultsReasons for switches to EVR were mainly malignancies before or after OLT, or chronic kidney injury. We were able to include 20 svLbx with EVR/lowTAC and 49 with highTAC/mycophenolate. Both groups had similar liver enzymes and similar kidney function. The EVR/lowTAC group exhibited lower TAC trough levels at svLbx (4.4 vs. 6.6 ng/ml; pDiscussionIn conclusion, EVR/lowTAC seems to control alloreactivity and histological graft injury as sufficiently as highTAC/mycophenolate within the first 3-4 years after OLT.</p

Image1_Combination of everolimus and low-dose tacrolimus controls histological liver allograft injury as sufficiently as high-dose tacrolimus.jpeg

IntroductionThe combination of everolimus (EVR) and low-dose tacrolimus (lowTAC) prevents T cell-mediated rejection of liver grafts as sufficiently as high-dose tacrolimus (highTAC) and mycophenolate, but is associated with a preserved kidney function within the first years after orthotopic liver transplantation (OLT). However, none of the available studies assessed the histological pattern of graft injury or fibrosis in surveillance biopsies (svLbx).MethodsAll svLbx taken under at least one month of stable immunosuppression with either EVR (aim 3-8 ng/ml) combined with lowTAC (aim 3-5 ng/ml) or highTAC (aim 5-8 ng/ml) combined with mycophenolate (500-1500 mg/day) within the first three to four years after OLT at our center were included. Patients who were switched to EVR because of insufficient control of alloreactivity were excluded.ResultsReasons for switches to EVR were mainly malignancies before or after OLT, or chronic kidney injury. We were able to include 20 svLbx with EVR/lowTAC and 49 with highTAC/mycophenolate. Both groups had similar liver enzymes and similar kidney function. The EVR/lowTAC group exhibited lower TAC trough levels at svLbx (4.4 vs. 6.6 ng/ml; pDiscussionIn conclusion, EVR/lowTAC seems to control alloreactivity and histological graft injury as sufficiently as highTAC/mycophenolate within the first 3-4 years after OLT.</p

Inhibition of HDV and HBV infection by P2XR antagonists and agonists is dependent on their charge. A

<p>) HepaRG cells were infected with serum of an HDV-positive patient (GTA7) in presence of P2XR antagonists (PPADS, suramin and AZ11645373) and agonists (ivermectin) for 16 h at 37°C. PreS/2-48^myr peptide (200 nM) was used in parallel as control. HDV-specific IF and nuclei staining was done 5 days p.i. The number of total and HDV-infected cells was quantified using ImageJ software. The results are depicted as percentage of the untreated control. In total, 290368 cells were counted for the analysis. Cell lysates of in parallel infected HepaRG cells were used for an HDAg-specific Western Blot. <b>B</b>) HepaRG cells were infected with HBV in presence of P2XR antagonists (PPADS, suramin and AZ11645373) and agonists (ivermectin) for 16 h at 37°C. Supernatants from days 8–11 p.i. were collected and secreted HBeAg was quantified by ELISA. Experiments were performed in duplicate and repeated at least two times independently. The values presented are mean ± SD. <b>C)</b> HepaRG cells were infected with serum of an HDV-positive patient (GTA7). Ivermectin (1 µM) was added at different time points during or after virus inoculation. HDV-specific IF and nuclei staining was done 6 days p.i. The number of total and HDV-infected cells was quantified using the ImageJ software. The results are depicted as percentage of the untreated control. In total, 128450 cells were counted for the analysis.</p

The obstruction or removal of negatively charged cellular interaction sites inhibit the HDV infection.

<p><b>A</b>) HepaRG cells were pre-incubated with increasing concentrations of poly-L-lysine for 30 min at 37°C and subsequently infected in presence of the polycation for 16 h at 37°C. <b>B</b>) HDV was pre-incubated for 1 h at 37°C with heparin (100 µg/ml and 500 µg/ml), suramin (100 µg/ml) and dextran sulfate (100 µg/ml). Unbound polyanions were removed by ultrafiltration. HepaRG cells were incubated with the pre-treated viruses for 8 h at 37°C. <b>C</b>) HepaRG cells were pre-incubated with heparin, suramin, dextran sulfate, pentosan polysulfate or poly-L-lysine for 1 h at 37°C. Cells were washed and incubated with HDV for 8 h at 37°C in absence of the compounds. <b>D</b>) Sulfation of cellular proteoglycans was inhibited by treatment of HepaRG cells with increasing concentrations of sodium chlorate for 48 h prior to HDV infection. <b>E</b>) HepaRG cells were incubated for 1 h at 37°C with the indicated concentrations of heparinase III or 1 µM preS/2-48^myr. The cells were washed and inoculated with HDV for 16 h at 37°C in presence of the substances. HDV-specific IF was performed for all experiments on day 6 p.i. Nuclei were stained with DAPI. The number of HDAg-positive cells and the total cell number were determined. The results are depicted as percentage of infected cells in comparison to the untreated control. In total, 822302 cells were counted for the analysis.</p

Biochemical, virological and immunological course of acute hepatitis C in asymptomatic patients with spontaneous clearance: patients with spontaneous clearance in the absence of anti-HCV

<p><b>Copyright information:</b></p><p>Taken from "Clearance of low levels of HCV viremia in the absence of a strong adaptive immune response"</p><p>http://www.virologyj.com/content/4/1/58</p><p>Virology Journal 2007;4():58-58.</p><p>Published online 11 Jun 2007</p><p>PMCID:PMC1914341.</p><p></p> HCV-RNA levels, ALT levels and T cell responses are shown. Coloured bars show the sum of SI values of the proliferation assay and the sum of specific interferon-gamma spots (spots in the presence of antigen – spots in the medium control) against 5 individual HCV proteins (HCV-core, HCV-NS3, HCV-helicase, HCV-NS4, HCV-NS5a). In addition, the number of individual proteins tested positive at the respective time point is shown for each assay. Interferon gamma ELISPOT responses were also tested against a panel of synthetic peptides representing class I and class II T cell epitopes derived from conserved regions of HCV with a cumulative HLA coverage > 85%. These assays were performed in an independent second laboratory ("Pep-Screen ELISPOT"; Vienna lab). The number of peptides tested positive in this assay is also given

Chemical structures, charge and function of the compounds used in the study.

<p>Listed are the structures of the repeating units (disaccharide or lysine) or the individual molecules. Charged groups are depicted in red. Possible acetylations are marked in blue. The number of charged groups per repeating unit or molecule and the function of the substance are listed. Not included are derivatives of the compounds.</p

Biochemical, virological and immunological course of acute hepatitis C in asymptomatic patients with spontaneous clearance: patients who developed anti-HCV antibodies

<p><b>Copyright information:</b></p><p>Taken from "Clearance of low levels of HCV viremia in the absence of a strong adaptive immune response"</p><p>http://www.virologyj.com/content/4/1/58</p><p>Virology Journal 2007;4():58-58.</p><p>Published online 11 Jun 2007</p><p>PMCID:PMC1914341.</p><p></p

Primers used for HDV detection.

<p>Primers used for HDV detection.</p

Phylogenetic analyses of HDV genomes in chronic HBsAg-positive patients in South Central Vietnam.

<p>(<b>a</b>) Representative HDV sequences of South Central Vietnamese HBV-HDV infection patients showing patient-specific HDV isolates. (<b>b</b>) Phylogenetic tree was inferred from distance analysis (Kimura 2 parameters model) and neighbor-joining reconstruction from HDV-region sequences. South Central Vietnamese HDV sequences are referred to as “letter/number”, i.e., “HM115”. The South Central Vietnamese HDV sequences were compared to HDV reference sequences, gathering the eight HDV clades (GenBank accession numbers are denoted in the figure). Phylogenetic analysis of HDV region nt 888 to nt 1122 showed that the HDV sequences were clustered in the Asian HDV-branches 1 and 5.</p

Baseline data and clinical characteristics of the HBV patients.

<p>Baseline data and clinical characteristics of the HBV patients.</p