Tadeusz Wojciech Łapiński

Update on alisporivir in treatment of viral hepatitis C

Introduction: There are two classes of anti-hepatitis C virus (HCV) agents currently in development: direct-acting antivirals (DAA) and host-targeting antivirals (HTA). Cyclophilin inhibitor alisporivir (ALV), previously known as Debio-025 is the most advanced HTA in development. Areas covered: Experimental and clinical studies demonstrated that ALV has high genetic barrier and no cross-resistance to DAA. Pharmacokinetic studies showed a profile suitable for once-daily administration. Phase I and II studies confirmed strong HCV suppression and that addition of ALV to pegylated IFNα (PegIFNα) and ribavirin (RBV) can improve their efficacy significantly. ALV was well tolerated and prevalence of the most frequent clinical and laboratory adverse events was similar to PegIFNα/RBV. Hyperbilirubinemia was the only significant adverse event related to ALV, but it was transient, reversible and not associated with hepatotoxicity or cholestasis. Expert opinion: ALV is pangenotypic, with once-daily administration and safe, therefore medication can be easy and flexible. There is still a need of data in difficult-to-treat populations and genetic studies allowing selection of possible non-responders. Registration of ALV for IFN-based treatment is expected within 3 years, but ALV is also a good candidate for IFN-sparing combinations with DAA.

HBV mutations and their clinical significance.

PURPOSE The launch of the vaccine against HBV in 1983 significantly reduced the number of HBV infections in the world. But there still remain a large group of people with chronic hepatitis B who were infected before beginning of vaccination programs or in whom the vaccine was - for various reasons - ineffective. Current therapy of HBV infection based on PEG-IFN α-2a or nucleotide/nucleoside analogues does not guarantee sustained virologic response in the large majority of chronically infected persons. Treatment with some nucleoside analogues is associated with mutations and subsequent selection of resistant strains resulting with therapeutic failure, risk of cross-resistance to other drugs and finally selection of mutants with oncogenic properties.

Plasma transforming growth factor β1, metalloproteinase-1 and tissue inhibitor of metalloproteinases-1 in acute viral hepatitis type B

AIM Antiproliferative, pro-apoptotic and immunosuppressive activity effects suggest crucial role of transforming growth factor (TGF)-beta1, metalloproteinase (MMP)-1 and its tissue inhibitor (TIMP)-1 in the pathogenesis of acute liver injury that in some patients precede development of chronic liver diseases and fibrogenesis. The aim of this study was to evaluate effect of acute HBV infection on plasma TGF-beta1, MMP-1 and TIMP-1 levels. METHODS TGF-beta1, MMP-1 and TIMP-1 plasma concentrations were measured with an enzyme immunoassay in 39 patients with acute viral hepatitis type B. Baseline measurement was performed within the first week of jaundice and then weekly up to the fourth week of the disease. Results were compared to baseline and normal values and to liver function tests. RESULTS Plasma concentrations of TGF-beta1, TIMP-1 and MMP-1 were significantly elevated in the first week of acute viral B hepatitis in comparison to normal. Analysis of individual values demonstrated significant positive correlation between plasma concentrations of TGF-beta1 and TIMP-1. There was no correlation between MMP-1 and TGF-beta1 or TIMP-1. Significant correlation was demonstrated between both TGF-beta1 and ALT or AST as well as between TIMP-1 and ALT, AST or bilirubin. Elevated baseline levels of both TGF-beta1 and TIMP-1 decreased gradually in consecutive weeks of the disease. TGF-beta1 but not TIMP-1 plasma concentrations were significantly lower in 3rd and 4th week than baseline values. MMP-1 concentration remained on baseline level in the 2nd week of the disease. However in the 3rd week its values increased suddenly but the significant difference in comparison to baseline was observed only in 4th week. CONCLUSIONS These results indicate important role of TGF-beta1, TIMP-1 and MMP-1 in acute viral hepatitis, that seems to be connected first of all with hepatocytes damage. Their role in extracellular matrix metabolism during acute liver injury needs further evaluation.

Helicobacter pylori infection among patients with liver cirrhosis

Background and aim Inflammatory changes in the stomach caused by Helicobacter pylori indirectly and directly affect liver function. Moreover, the bacteria may worsen the course of the liver cirrhosis. The study aimed at evaluating the incidence of H. pylori infection among patients with liver cirrhosis, depending on the etiology and injury stage, scored according to Child–Pugh classification. Stage of esophageal varices and endoscopic inflammatory lesions in the stomach were evaluated, depending on the presence of H. pylori infection. Patients and methods The study included 147 patients with liver cirrhosis: 42 were infected with hepatitis C virus, 31 were infected with hepatitis B virus, 56 had alcoholic liver cirrhosis, and 18 had primary biliary cirrhosis. Diagnosis of H. pylori infection was performed based on the presence of immunoglobulin G antibodies in serum. Results H. pylori infection was found in 46.9% of patients. The incidence of H. pylori infection among patients with postinflammatory liver cirrhosis was significantly higher (P=0.001), as compared with patients with alcoholic liver cirrhosis. Ammonia concentration was significantly higher in patients infected with H. pylori, compared with noninfected individuals (129 vs. 112 &mgr;mol/l; P=0.002). Incidence of H. pylori infection in patients without esophageal varices was significantly lower compared with patients with esophageal varices (14 vs. 60%; P<0.001). Conclusion H. pylori infection is significantly more frequent among patients with postinflammatory liver cirrhosis (infected with hepatitis C virus or hepatitis B virus) than in patients with alcoholic liver cirrhosis or primary biliary cirrhosis. H. pylori infection correlates with elevated concentration of blood ammonia and the incidence of esophageal varices.

Significance of mutations in the region coding for NS3/4 protease in patients infected with HCV genotype 1b.

BACKGROUND Fast hepatitis C virus (HCV) replication is one of the reasons for frequent changes in viral genome. OBJECTIVES The objective of this study was to evaluate the frequency and type of mutation in NS3/4 protease in patients with HCV genotype 1b and to determine the effect of the mutation on viral load, fibrosis stage, alanine aminotransferase (ALT) activity, and alpha-fetoprotein (AFP) level. MATERIAL AND METHODS The study included 46 treatment-naïve patients, infected with HCV genotype 1b. Mutations were analyzed after isolating HCV RNA, and then evaluating the compliance of the amino acid sequence, using 3500 Genetic Analyzer (Applied Biosystems, Foster City, USA). RNA fragment from nucleotide 1-181 encoding NS3/4 protease was subjected to analysis. RESULTS Mutations were demonstrated in 65% of subjects. Changes in the protease region affecting resistance to treatment (T54, Q80, V158, M175, D186) were detected in 10.8% of patients. Substitution mutation at T72 was found most frequently - in 49.9% of cases. In 13% of patients, mutation at G86 was demonstrated, including G86P in 5 patients and G86S in 1 patient. In the group of patients with T72 mutation, viral load was significantly higher (1.3 × 106 IU/mL vs 1.0 × 105 IU/mL; p = 0.01), AFP level was higher and fibrosis level was lower (1.26 vs 2.17; p = 0.008) compared to the patients without the mutation. Cryoglobulinemia was observed in 74% of patients with mutation at position T72. CONCLUSIONS Natural mutations of the region coding for NS3/4 protease are found frequently in patients infected with genotype 1b, but they may cause resistance to antiviral agents only in 11% of patients. Changes were most frequently found at position T72. Mutations at position T72 are correlated with the cryoglobulinemia occurrence. This is a substitution mutation, accompanied by a high viral load, high ALT activity and AFP level, which may point to a more unfavorable influence of such a modified virus, compared to wild-type virus, onto pathological processes in the liver.

The occurrence of autoantibodies in patients with chronic HCV infection, including patients dialyzed and after kidney transplantation

Introduction There are reports suggesting that hepatitis C virus (HCV) may stimulate the autoimmune process. Studies have been undertaken to evaluate the occurrence and type of autoantibodies in HCV-infected patients with and without immunosuppression. Results were analyzed according to HCV genotype, intensity of inflammation and liver fibrosis stage. Material and methods The study included 105 patients chronically infected with HCV, including 25 with immunological suppression administered for kidney disease or kidney transplantation. Blood samples were tested by immunoblotting for the presence of AMA-M2, SLA/LP, LKM-1, LC1, anti-F-actin, anti-desmin, anti-myosin, anti-gp210 and anti-sp100 autoantibodies, and ANA. All the patients were scored for autoimmune hepatitis. Results Autoantibodies were detected in 32.5% of patients without immunosuppression and in 16% with immunosuppression. Single types of autoantibodies were identified in 26% of patients. The most frequent ones were ANA (19%) and AMA-M2 (5.7%). The presence of antibodies in patients with genotype 1 was significantly higher in comparison to their occurrence in genotype 3. Autoimmune hepatitis was not diagnosed in any of the patients. Immunoglobulin G level was significantly higher in patients with detectable autoantibodies, compared to patients without antibodies (1.89 vs. 1.28 g/dl, p < 0.001). No correlation between fibrosis stage or intensity of inflammatory state and the frequency of antibodies was found. Conclusions The antibodies are significantly more frequent in patients without immunosuppression and in patients infected with genotype 1 than genotype 3. The presence of these autoantibodies is not associated with the development of autoimmune hepatitis. Higher level of immunoglobulin G in the serum correlates with the presence of autoantibodies.

Effect of hepatitis B virus (HBV) infection on the course of pregnancy and newborns’ health status

Introduction The effect of chronic hepatitis B virus (HBV) infection on pregnancy is not clear. Hepatitis B virus infection of newborns in the case of natural delivery occurs in 70-90% of cases. Risk factors of infection are the presence of serum HBeAg and HBV DNA level above 107 IU/ml. Active and passive prevention protect more than 95% of neonates born to mothers infected with HBV. The aim of the study was to determine the course of pregnancy in HBV-infected women, the mode of delivery, efficacy of prophylaxis against HBV infection in newborns, and health condition of newborns within the first years of life. Material and methods The course of 104 pregnancies in 69 women infected with HBV was monitored. Hepatitis B virus viral load was determined by PCR using the AmpliPre/COBAS TaqMan HBV system. Neonatal HBV infection and the health condition at birth and during the first year of life were analyzed. Results All included pregnant women were HBeAg negative. No clinically significant disorders were observed during pregnancy. Viral load measured in the third trimester did not exceed 107 IU/ml in any pregnant woman. Only 5 (8%) of them demonstrated levels above 105 IU/ml. Two women (1.9%) experienced a miscarriage, which was considered as not associated with HBV infection. The majority (56%) of pregnancies ended with spontaneous labor. Complete prevention against HBV was applied in 79% of newborns. Hepatitis B virus infection was diagnosed in 3 children who received incomplete or no prophylaxis. Hepatitis B virus infection occurred in 3 (2.9%) children born naturally, who did not receive proper prevention after delivery. The Apgar score in children born to mothers infected with HBV did not differ significantly from that in neonates born to healthy women from the same population. Allergic disorders developed in 17 children who underwent HBV prophylaxis. Conclusions Low viral load in pregnant women infected with HBV and the absence of HBeAg reduce the probability of infection of newborns. Proper prevention carried out after delivery seems to be the most important method to prevent HBV infection in newborns.

Original article Impact of rs12979860 polymorphism on liver morphology in chronic HCV infection

Aim of the study To determine distribution of rs12979860 genotypes, their correlations with viral load as well as inflammatory activity and stage of liver fibrosis in patients infected with HCV genotype 1. Material and methods The study included 132 patients infected with HCV genotype 1b. Serum viral loads were obtained with the PCR method. Rs12979860 polymorphisms were determined by sequencing of PCR products. Liver biopsy was performed in all patients. Results CT, TT and CC alleles of rs12979860 polymorphism were detected in 58%, 20% and 22% of patients respectively. The highest viral load was observed in the TT and the lowest in the CC group (72.0 × 106 IU/ml vs. 2.1 × 106 IU/ml, p < 0.005). A significant correlation was demonstrated between patient’s age and inflammatory activity as well as degree of liver fibrosis. No association was found between liver histopathology and HCV viral load or rs12979860 genotypes. Conclusions There is an association between HCV viral load and rs12979860 polymorphism. Inflammatory activity and stage of liver fibrosis depend on age, but there is no relationship with rs12979860 genotypes and HCV viral load.

Wyniki leczenia przewlekłego zapalenia wątroby typu B entekawirem

Hepatitis B virus is a main cause of chronic hepatitis regarding 350 million people worldwide, leading to life-threatening conditions: liver cirrhosis and hepatocellular carcinoma. The worsening progress may be blocked or delayed by the effective therapy. Entecavir is an oral nucleoside analogue, safe and well tolerated, very effective inhibitor of HBV polymerase resulting in rapid viral suppression, histological improvement and ALT normalization. In several randomized, multicentre trials entecavir was effective in nucleoside-naïve and lamivudinerefractory patients, hepatitis B e antigen positive or negative as well as patients with decompensated liver cirrhosis.

Treatment of chronic hepatitis C in a patient with Fanconi anaemia

Fanconi anaemia is a rare autosomal recessive disorder with progressive bone marrow failure and predisposition to malignancy. We report a case of a 26-year-old female patient with Fanconi anaemia and severe chronic active hepatitis C virus infection. Her past medical history included treatment with multiple blood transfusions and bone marrow transplantation at the age of 13. The decision to treat the infection was taken, and history of hematologic disease contributed to the introduction of therapy with leukocyte interferon-α n3 and ribavirin combined with a granulocyte - colony stimulating factor. The treatment was well tolerated and after 48 weeks a reduction of the viral load and alanine aminotransferase activity were achieved. No adverse effects on bone marrow functioning were noted.