Preclinical evaluation of two human anti-hepatitis B virus (HBV) monoclonal antibodies in the HBV-trimera mouse model and in HBV chronic carrier chimpanzees

Two human monoclonal antibodies (mAbs) against hepatitis B surface antigen (HBsAg) generated in the Trimera mouse system are described. Both mAbs 17.1.41 and 19.79.5 are of the IgG1 isotype and have high affinity constants for HBsAg binding in the range of 10(-10) mol/L. Monoclonal antibody 17.1.41 recognizes a conformational epitope on the a determinant of HBsAg whereas mAb 19.79.5 recognizes a linear one. The 2 mAbs bind to a panel of hepatitis B virus (HBV) subtypes with distinct patterns. The neutralizing activity of these antibodies was tested in 2 different animal model systems. Administration of each mAb to HBV-Trimera mice, a system that provides a mouse model for human hepatitis B infection, reduced the viral load and the percentage of HBV-DNA-positive mice in a dose-dependent manner. These 2 mAbs were more effective than a polyclonal antibody preparation (Hepatect; Biotest Pharma, Dreieich, Germany) in both inhibition of HBV liver infection and reduction of viral load. A single administration of a mixture of these mAbs into HBV chronic carrier chimpanzees resulted in immediate reduction in HBsAg levels followed by recurrence to initial levels within few days. Thus, these mAbs may be potential candidates for preventive therapy or in combination with other antiviral agents against HBV. Further studies in humans are needed to assess these mAbs in various clinical indications.     

Severe aplastic anemia associated with seronegative community-acquired hepatitis C virus infection

Summary Aplastic anemia (AA) is a rare complication of viral hepatitis affecting mainly children or young adults. Most reported cases have been associated with community-acquired non-A, non-B hepatitis, but hepatitis A and B have also been implicated in a few patients. We report on a 43-year-old woman with severe AA, in association with acute hepatitis C virus (HCV) infection, diagnosed by detection of HCV RNA by the polymerase chain reaction technique. Her AA was successfully treated with anti-thymocyte globulin and cyclosporin A. The hepatitis C progressed to chronic disease but, despite a follow-up time of 2 years, the patient still has no detectable anti-HCV antibodies, as evaluated with a secondgeneration anti-HCV assay.     

Late seroconversion to hepatitis B in a Somali village indicates the important role of venereal transmission.

Hepatitis B virus (HBV) markers were investigated in a cross-sectional study in 1985 on sera from 84% of the 648 inhabitants in a rural Somali village. The prevalence of HBV markers increased with age, from 9.7% in subjects less than 12 years old, to 38% in the age group 12-19 years, and to 68% in adults. HBV markers were more common in boys less than 12 years old, 13%, than in girls of the same age group, 5.8% (P less than 0.05). A rapid increase of HBV markers started at adolescence in both sexes. The female cohort showed their highest seroconversion rate during their second decade of life, while the male cohort seroconverted more rapidly in the third decade. Thus, an initially more rapid seroconversion among boys was reversed when the females reached reproductive age, and no sex difference in marker frequencies was observed in the age group 12-19 years. There was a steady increase of HBV markers during the reproductive years in both sexes. The frequencies of HBsAg, as well as total markers, were significantly higher in adult males than females, 14 vs 5.6%, and 77 vs 62%, respectively. HBV markers were more frequent in wives of HBV positive husbands than in those married to HBV negative husbands. No increased marker prevalence was observed among siblings of HBV positive children, nor among their mothers, which disproved the role of vertical and early horizontal transmission. In 1989 the four-year rate of seroconversion was investigated in villagers who were seronegative in 1985. On testing 158 sera from 319 individuals, the seroconversion rate was significantly lower among those younger than 12 years in 1985 compared to those in the older age group, 5 vs 17%.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sequential changes in hepatitis A virus genotype distribution in Estonia during 1994 to 2001

Hepatitis A virus (HAV) isolates from a large outbreak and from non-outbreak cases in Estonia were characterized by sequencing the aminoterminal VP1 region. From January 1998 to December 1999, a total of 1084 cases of hepatitis A were reported to the Harjumaa-Tallinn and Ida-Virumaa Health Protection Services in Estonia. The attack rate was highest among males aged 15-29. Initial cases were noted to be associated with injecting drug use. IgM anti-HAV positive sera were available from 107 hospitalized outbreak cases and from 68 patients sampled during 1994 to 2001. HAV RNA was detected in 42% of sera from 1994-1996 and in 88% of sera from 1998-2001. It was possible to obtain HAV sequences from 83 outbreak and 29 background cases. The outbreak strain was represented by five different sequences, all belonging to subtype IIIA. During the outbreak, this IIIA strain also spread into the general population. All available non-outbreak isolates from 1994 to 2001 but one belonged to genotype IA and formed distinct clusters as compared to isolates from other parts of the world. One subtype IIIA isolate from 1995 was unrelated to the outbreak strain. Subtype IA had been dominating in Estonia during 1994-2001, but the outbreak strain from 1998 to 1999 was IIIA. This subtype was encountered previously in addicts in Sweden during the 1980s and in Norway at the end of the 1990s. This study supports the use of limited sequencing within the aminoterminal VP1 region for studying the molecular epidemiology of hepatitis A.     

Confirmation of Nosocomial Transmission of Hepatitis C Virus by Phylogenetic Analysis of the NS5-B Region

Four hepatitis C virus transmission chains at three dialysis units were disclosed by limited sequencing; three of these were disclosed by analysis of the NS5-B region of the genome. Dialysis on the same shift as that during which infected patients were dialyzed was the common factor for seven patients in two chains. Two nurses exposed to needle sticks and their sources of infection constituted two other chains. The strains of three chains belonged to subtype 1a and formed clusters with an intrachain variability of 0 to 6 nucleotides compared to 8 to 37 nucleotides for unrelated strains within this subtype. The clusters were supported by bootstrap values ranging from 89 to 100%.     

Patient-to-Patient Transmission of Hepatitis C at Iranian Thalassemia Centers Shown by Genetic Characterization of Viral Strains

Background

Hepatitis C is prevalent among thalassemia patients in Iran. It is mainly transfusion mediated, in particular among patients treated before 1996 when blood screening was introduced.

Objectives

The current study aimed to investigate why patients still seroconvert to anti-HCV in Iranian thalassemia centers.

Patients and Methods

During 2006-2007 sera were sampled from 217 anti-HCV positive thalassemia patients at nine thalassemia centers in Tehran and Amol city, where 34 (16%) patients had been infected after 1996. The HCV subtype could be determined by sequencing and phylogenetic analysis of partial NS5B and/or 5׳NCR-core region in 130 strains.

Results

1a (53%) was predominant followed by 3a (30%), 1b (15%), and one strain each of 2k, 3k and 4a. Phylogenetic analysis revealed 19 clades with up to five strains diverging with less than six nucleotides from each other within subtypes 1a and 3a. Strains in seven clades were from nine patients infected between 1999 and 2005 and similar to strains from eight patients infected before 1996, indicating ongoing transmission at the centers. Further epidemiological investigation revealed that 28 patients infected with strains within the same clade had frequently been transfused at the same shift sitting on the same bed. An additional eight patients with related strains had frequently been transfused simultaneously in the same room.

Conclusions

The results suggest nosocomial transmission at these thalassemia centers both before and after the introduction of blood screening. Further training of staff and strict adherence to preventive measures are thus essential to reduce the incidence of new HCV infections.     

Locations of antibody binding sites within conserved regions of the hepatitis C virus core protein

The binding sites for human antibodies recognising antigenic domains within the hepatitis C virus (HCV) core protein were analyzed using synthetic peptides. Omission peptide analogues where a pair of residues was sequentially omitted were produced corresponding to the regions 1–18, 11–28, 21–38, 51–68, and 101–118. The N-terminal part of HCV core was found to contain three distinct antibody binding sites, which includes the previously reported one at residues 9–16. The other two were located at residues 19–26 and residues 29–34. Within the region 51–68 two overlapping sites were found, the first at residues 51–60 and the second at residues 59–68. Within the region 101–118, residues 107–114 were identified as the binding site, which contains two residues differing between genotypes I/II and III/VI. Thus the HCV core protein contains at least six distinct linear antibody binding sites, located at regions highly conserved between the genotypes of HCV. The human recognition of these regions show a variation with respect to the amino- and carboxy-terminal extension of each individual binding site. These findings will have implications for the prediction of the structure of the HCV core protein, since these antibody binding sequences are likely to be more or less accessible from the exterior of either, or both, of the native HCV core and its precursor polyprotein. © 1994 Wiley-Liss, Inc.     

Late seroconversion and high chronicity rate of hepatitis C virus infection in patients with hematologic disorders

BACKGROUND: Patients with hematologic disorders requiring repeated bloodand platelet transfusions are at high risk for development ofpost-transfusion non-A, non-B hepatitis. PATIENTS AND METHODS: Fifty-five patients with hematologic diseases and post-transfusionnon-A, non-B hepatitis were studied. Sera were assayed for hepatitisC virus (HCV) antibodies with a second-generation enzyme-linkedimmunoassay. Sera from 40 patients were examined for the presenceof HCV RNA with a nested PCR method. RESULTS: The clinical picture of acute non-A, non-B hepatitis did notdiffer from that described in other patient groups: however,progression to chronic hepatitis was very common (95%). Thirty-eight(95%) of 40 patients, whose sera were analysed both serologicallyand for the presence of HCV RNA had verified HCV infections.In some patients time to seroconversion was prolonged, up tomore than 14 months. Seventeen patients with resistant or relapsedacute leukemia were treated with combination chemotherapy duringthe acute or chronic phase of hepatitis. Suppression of theinflammatory activity as reflected by a decrease of serum aminotransferaselevels was recorded during the subsequent pancytopenic period. CONCLUSIONS: Hepatitis C has a high chronicity rate in patients with hematologicdisorders which parallels the situation of hepatitis B in theimmunocompromised host. Furthermore, like the situation in hepatitisB, the hosts' immune response to infection seems to be involvedin the pathogenesis of liver injury. Time to seroconversionmay be prolonged and detection of HCV RNA is therefore importantfor diagnosis. immunosuppression, hematologic diseases, hepatitis C, chronic active hepatitis, non-A and non-B hepatitis     

3 year simvastatin treatment and lens nuclear back scattering

AIM: To determine if 3 year treatment of hypercholesterolaemia with simvastatin causes an increase of lens nuclear back scattering. METHODS: 160 patients with hypercholesterolaemia in the Scandinavian Simvastatin Survival Study (4S) were followed for 3 years. Half (80) of the patients took simvastatin and half (80) received placebo. The lens was photographed with a Topcon SL-45 slit lamp camera at the beginning and at 1 year intervals. A common lens nuclear area was used for measuring lens nuclear back scattering. RESULTS: Nuclear back scattering increased with age and there was more pronounced scattering in women than in men. Lens nuclear back scattering did not differ significantly between the simvastatin and placebo groups, but the power was low (0.2). Lens nuclear back scattering increased during the study period independently of baseline back scattering, age, and sex for both groups. CONCLUSION: Although no significant difference was found between the simvastatin and placebo groups, the currently available data are insufficient for exclusion of the possibility that taking simvastatin during a 3 year period increases nuclear back scattering. However, a possible minor increase of nuclear back scattering is clinically irrelevant considering known beneficial effects of simvastatin on coronary heart disease.