Hepatitis B surface antigen reduction by switching from long-term nucleoside/nucleotide analogue administration to pegylated interferon.

Hepatitis B surface antigen (HBsAg) reduction during nucleoside/nucleotide analogue (NA) therapy is slow and an alternative strategy for patients receiving ongoing NA to facilitate HBsAg reduction is required. We investigated whether switching to pegylated interferon (PEG-IFN) after long-term NA administration enhances HBsAg reduction. Forty-nine patients who switched from long-term NA to 48 weeks of PEG-IFN alfa-2a were studied. The mean duration of previous NA was 48 months (sequential group). A total of 147 patients who continued NA and matched for baseline characteristics were analysed for comparison (NA continuation group). The treatment response was defined as HBsAg reduction ≥1.0 logIU/mL at the end of PEG-IFN. HBsAg reduction at week 48 was 0.81±1.1 logIU/mL in the sequential group, which was significantly higher than that in the NA continuation group (0.11±0.3 logIU/mL, P < .001). The treatment response was achieved in 29% and 2% of the sequential group and NA continuation group (P < .001), and the odds ratio of sequential therapy for the treatment response was 19 compared with the NA continuation (P < .001). In patients tested positive for hepatitis B e antigen (HBeAg), HBeAg seroconversion was higher in the sequential group (44% vs 8%, P < .001). In HBeAg-negative patients, only patients in the sequential group achieved HBsAg loss. No patient needed to resume NA administration because of HBV DNA increase accompanied by alanine aminotransferase flares. In summary, sequential therapy with PEG-IFN after long-term NA enhances the reduction of HBsAg and may represent a treatment option to promote HBsAg loss.

ITPA genetic variants influence efficacy of PEG-IFN/RBV therapy in older patients infected with HCV genotype 1 and favourable IL28B type.

Inosine triphosphatase (ITPA) genetic variants are strongly associated with ribavirin (RBV)-induced anaemia during pegylated interferon (PEG-IFN) plus RBV therapy. However, the treatment efficacy of ITPA genetic variants has not been fully explored. We enrolled 309 individuals infected with hepatitis C virus genotype 1, who were treated with PEG-IFN plus RBV for 48 weeks. The ITPA SNP: rs1127354 and IL28B SNP: rs8099917 were genotyped. We examined the risk factors for severe anaemia up to week 12 after the start of treatment and treatment efficacy. The incidence of severe anaemia, ≥ 3 g/dL reduction or <10 g/dL of haemoglobin (Hb) up to week 12, was more frequent in patients with CC at rs1127354 [65% (145/224), 33% (73/224)] than in those with CA/AA [25% (21/85), 6% (8/85)] (P < 0.0001). ITPA genotype, pretreatment Hb level and age were independent predictive factors for severe anaemia: Hb < 10 g/dL. In IL28B favourable type, the sustained virologic response rate was higher in ≥ 60-year-old patients with CA/AA than in those with CC [71% (22/31) vs 40% (26/65), P = 0.005], although there was no significant difference in treatment efficacy according to ITPA genetic variants in the <60-year-old patients. The proportion of patients administered ≥ 80% of the dosage of RBV was significantly higher in the patients with CA/AA than in those with CC (P = 0.025), resulting in a lower relapse rate. In conclusion, ITPA genetic variants were associated with severe RBV-induced anaemia and could influence the efficacy of PEG-IFN plus RBV treatment among elderly patients with IL28B favourable type.

Mechanism of the dependence of hepatitis B virus genotype G on co-infection with other genotypes for viral replication.

Hepatitis B virus (HBV) is classified into several genotypes. Genotype G (HBV/G) is characterised by worldwide dispersion, low intragenotypic diversity and a peculiar sequence of the precore and core region (stop codon and 36-nucleotide insertion). As a rule, HBV/G is detected in co-infection with another genotype, most frequently HBV/A2. In a previous in vivo study, viral replication of HBV/G was significantly enhanced by co-infection with HBV/A2. However, the mechanism by which co-infection with HBV/A2 enhances HBV/G replication is not fully understood. In this study, we employed 1.24-fold HBV/A2 clones that selectively expressed each viral protein and revealed that the core protein expressing construct significantly enhanced the replication of HBV/G in Huh7 cells. The introduction of the HBV/A2 core promoter or core protein or both genomic regions into the HBV/G genome showed that both the core promoter and core protein are required for efficient HBV/G replication. The effect of genotype on the interaction between foreign core protein and HBV/G showed that HBV/A2 was the strongest enhancer of HBV/G replication. Furthermore, Western blot analysis of Dane particles isolated from cultures of Huh7 cells co-transfected by HBV/G and a cytomegalovirus (CMV) promoter-driven HBV/A2 core protein expression construct indicated that HBV/G employed HBV/A2 core protein during particle assembly. In conclusion, HBV/G could take advantage of core proteins from other genotypes during co-infection to replicate efficiently and to effectively package HBV DNA into virions.

Effector mechanism in rejection of allografts expressing an isolated minor histocompatibility disparity. Importance of cytotoxic T lymphocytes in the rejection of H-43a allografts by H-43b mice.

To explore effector mechanisms in allograft rejection, we transplanted skin grafts (SG) across a single minor histocompatibility locus (H-43) using mouse strains carrying the H-43b allele as SG recipients and those carrying the H-43a allele as SG donors. Recipients' spleen cells (SC) were assayed at various intervals for 200 days for anti-H-43a cytotoxic T lymphocyte (CTL) responsiveness, as well as delayed-type hypersensitivity (DTH) responsiveness. When H-43a SG from C3H.SW mice were transplanted to H-43b CWB mice, two thirds of the recipients rejected the SG, and recipients' SC showed marked self-H-2Kb-restricted anti-H-43a CTL responsiveness until the end of the observation period. In contrast,H-43aSG transplanted to H-43b (B10.BRxCWB)F1 (BWF1) mice survived in almost all of the BWF1 recipients. The anti-H-43a CTL responsiveness of the recipients' SC was evident until day 40 but thereafter started to wane and eventually disappeared. Notably, BWF1 mice whose self-H-2Kb-restricted anti-H-43a CTL precursors had been primed by prior injection with H-43a SC rejected H-2Kb-bearing H-43a CSW SG but not H-2k, H-43a C3H/HeN SG. In contrast, an anti-H-43a DTH response was not induced in any of the CWB and BWF1 recipients, including CWB recipients who rejected the H-43a SG. Since it has been well documented that anti-H-43a CTL are restricted solely by self-H-2Kb, the results in this study indicate that self-H-2Kb-restricted anti-H-43a CTL are responsible for rejection of H-43a allografts by H-43b recipient mice.

Localisation of intrahepatic interleukin 6 in patients with acute and chronic liver disease.

AIM: To evaluate the role of local interleukin 6 (IL-6) in the pathogenesis of acute and chronic liver disease. METHODS: The cellular site of IL-6 in cryostat sections of liver from 31 patients with liver disease was examined using indirect immunofluorescence with a monoclonal antibody. RESULTS: IL-6 staining in sinusoidal endothelial cells was very noticeable and diffusely distributed in the lobules of specimens of acute viral hepatitis. IL-6 expression in endothelial cells, particularly in necrotic areas of hepatocytes, was increased and was accompanied by enhanced expression in Kupffer cells. In contrast, IL-6 staining in infiltrating mononuclear cells was prominent in portal tracts, and the numbers of cytokine positive cells were greater in specimens of chronic active hepatitis compared with chronic persistent hepatitis. In non-specific reactive hepatitis intrahepatic expression of IL-6 was minimal, while in alcoholic liver fibrosis the cytokine distribution in the lobules was similar to that of acute viral hepatitis. CONCLUSIONS: These results indicate that locally produced IL-6 contributes to the inflammatory process and immunological response in acute and chronic liver disease.

Randomized controlled trial of twice-a-day administration of natural interferon beta for chronic hepatitis C.

We conducted a randomized controlled trial to assess the efficacy of twice-a-day administration of natural interferon beta (IFNbeta) as an induction of IFN therapy for chronic hepatitis C. Seventy-one patients with chronic hepatitis C were enrolled into the trial and randomly assigned into three treatment groups. Six million units (MU) of IFNbeta were administered once-a-day for the first 4 weeks, and then thrice weekly for 12 weeks in 20 patients (once-a-day group). Three milion units of IFNbeta were administered twice-a-day for the first 2 weeks, 6 MU once-a-day for the next 2 weeks, and then thrice weekly for 12 weeks in 23 patients (twice-a-day+beta group), or 6 MU of lymphoblastoid IFNalpha were administered thrice weekly for the last 12 weeks instead of IFNbeta in 28 patients (twice-a-day+alpha group). Four patients in once-a-day group (20%), 9 in twice-a-day+beta group (39%), and 12 in twice-a-day+alpha group (43%) obtained sustained response. Sustained response rate in twice-a-day groups was higher than in once-a-day group, although there was no statistical significance. The present study suggested the possible superiority of twice-a-day administration of IFNbeta as an induction therapy to once-a-day administration, but further studies are needed to confirm this regimen.

Percutaneous transhepatic balloon dilation for papillary stenosis.

In this study, a case of primary common bile duct stone due to papillary stenosis is reported. The patient was a 68-year-old man with complaints of epigastric pain and fever who had undergone gastrectomy at age 55 and cholecystectomy with choledocholithotomy at age 62. Laboratory data revealed elevation of the transaminases acid biliary enzymes. Both abdominal ultrasonography and CT scan revealed dilatation of the common bile duct with stones. Since endoscopic retrograde cholangiopancreatography could not visualize the bile duct, percutaneous transhepatic biliary drainage was carried out. After lithotripsy by percutaneous transhepatic cholangioscopy a diagnosis of papillary stenosis was made following percutaneous transhepatic manometry of the sphincter of Oddi, and balloon dilatation through the PTBD fistula was successfully performed. In this case report, emphasis is placed on the diagnosis and treatment of papillary stenosis.

Subchondral cancellous bone in osteoarthrosis and rheumatoid arthritis of the femoral head. A quantitative histological study of trabecular remodelling.

Thirty-three femoral heads taken from patients undergoing total hip replacement for osteoarthrosis and rheumatoid arthritis have been examined in undecalcified, plastic-embedded sections, using a quantitative histological methods. The characteristic change of trabecular remodelling in the subchonfral cancellous bone of the femoral heads was that total osteoid surface in the medial osteoarthritis (medial O.A.) and rheumatoid arthritis (R.A.) was significantly less than in the proximal osteoarthrosis (proximal O.A.) (P less than 0.01). Resorption surface was slightly greater in R.A. than in proximal O.A. (P less than 0.05). Bone area in the deep area was not different in each group. In the superficial area it was significantly less in medial O.A. than in proximal O.A. (P less than 0.05). The average width of trabeculae was slightly less in medial O.A. and R.A. It is suggested that subchondral trabecular remodelling may be a basic pathological process of general importance in the evolution of these diseases.

Quantitative ultrastructural analyses of rabbit osteoclasts. The effect of in vivo treatment with sodium salicylate.

Rabbit tibial osteoclasts were examined by electron microscopy and quantitative data on their ultrastructural morphology collected by methods described by Holtrop. Osteoclasts from the tibiae of two groups of rabbits were compared: one fed a commercial diet and the other fed the same diet containing 2% sodium salicylate (v/w). No changes in the total number of tibial osteoclasts were detected but average osteoclast size, numbers of nuclei per osteoclast and ruffled border and clear zone areas decreased (p less than 0.05), as did the proportion of osteoclasts directly attached to bone. These results suggest that osteoclast activity is inhibited by in vivo salicylate therapy.

Cytotoxic T lymphocyte responses to minor H-43 alloantigens in H-43a and H-43b mice. Both anti-H-43b and anti-H-43a CTL activities are generated exclusively in the context of the same H-2Kb restriction element.

We have previously demonstrated that anti-H-43a CTL response of H-43b responder mice was exclusively restricted by self H-2Kb (Kb) but not by the other nine self MHC class I alleles from independent origins, i.e., Kbml,d,k,s and Db,d,k,q,s. In the present study, we verified that Kf,q,r and Df,r alleles could also not serve as restricting class I elements in the CTL response to H-43a alloantigen. Another notable observation made in the earlier study was the fact that, in H-43 incompatibility of the alternative combination, H-43a mice were incapable of generating CTL activity against H-43b alloantigen. However, by means of employing new in vivo immunization procedures, we discovered that some but not all genetically identical H-43a responder mice could mount anti-H-43b CTL response restricted by self Kb. Again, no anti-H-43b CTL activity could be generated in the context of self Kk, Kj, Db or Dk molecules. Although the number of class I alleles we examined is still limited, these results indicate that antigenic fragments derived from the processed H-43a and H-43b alloantigens possess an indistinguishable epitope (agretope), and that such agretope either interacts only with the privileged Kb molecules or allows to bestow the immunogenic conformation of allodeterminants on the fragments solely in the context of the restricting Kb element.

Plasmids in Vibrio parahemolyticus strains isolated in Japan and Bangladesh with special reference to different distributions.

We surveyed plasmids in naturally occurring Vibrio parahemolyticus strains isolated in Japan and Bangladesh. Among the strains isolated in Japan, about half of the strains isolated from stools of patients of domestic diarrhea outbreaks as well as of travelers returning from East Asia were found to have plasmids, but no strains from foods had plasmids. In contrast, among the strains isolated in Bangladesh, none of the four strains isolated from patients had plasmids, but two out of eight strains isolated from water had plasmids, suggesting that plasmids are common in strains from the water in Bangladesh. All plasmids so far reported in V. parahemolyticus were detected in strains isolated from stools of patients. Incidences of plasmids in this organism were not so high in either area. In Japan, all plasmids were detected in strains from human intestines at 37 C, but in Bangladesh, where the temperature is around 30-40 C, the plasmids were detected in strains from the natural environment. These results suggested the possibility that these plasmids can come from different bacteria under rather high temperatures and that incidences of plasmids are influenced by the incidences of plasmids in bacteria present in the vicinity of V. parahemolyticus strains. None of these plasmids were found to have any relation to the biological characters tested.

Long-term carriage of hepatitis C virus with normal aminotransferase after interferon treatment in patients with chronic hepatitis C.

Studies were undertaken to investigate whether interferon therapy could induce hepatitis C virus (HCV) carriage with normal serum alanine aminotransferase (ALT) values using an assay that combined reverse transcription and polymerase chain reaction. The subjects studied were 53 patients with chronic active hepatitis C who received interferon (alpha, 33 cases; beta, 20 cases) therapy. All were seropositive for HCV RNA prior to therapy. In all 22 complete responders, whose ALT levels fell to normal during therapy and for at least 24 weeks after therapy, HCV RNA became persistently negative except in two cases. The two had sustained viremia on treatment and for 1.0-1.5 years of follow-up, although their biochemical tests were normal. In 15 patients with a transient response in whom the disease recurred when interferon was stopped, HCV RNA was undetectable in 80% of the cases at the end of therapy, but the virus reappeared with subsequent elevation of ALT in all patients. However, 3 patients in this group had normal enzyme levels with viremia for 2.1-2.8 years of follow-up after acute deterioration of illness. In 16 patients who did not respond to interferon, HCV RNA was persistently positive during and after therapy. These findings suggest that interferon therapy induces a long-term carrier state of HCV infection with normal ALT levels in some patients.