Liver iron accumulation in chronic hepatitis C patients without HFE mutations: relationships with histological damage, viral load and genotype and alpha-glutathione S-transferase levels.

BACKGROUND: Host and viral factors have been suggested as possible causative factors for the presence of liver iron accumulation in chronic hepatitis C. However, there is no agreement regarding the influence of liver iron accumulation on the biochemical and histological severity of chronic hepatitis C. Moreover, data concerning the relationships between both viral load and genotype and liver iron accumulation are scanty. AIMS: To evaluate the biochemical, histological and virological assessment of a group of chronic hepatitis C patients without risk factors for iron overload, on the basis of the presence, degree and distribution of liver iron accumulation. METHODS: Fifty-three chronic hepatitis C patients (34 men, 19 women; age 44 +/- 11 years) with no risk factors for liver iron accumulation and showing no HFE mutations were chosen from a broader cohort of chronic hepatitis C patients. The presence, degree and distribution of liver iron accumulation were assessed using Deugnier's score. Relationships between the presence of liver iron accumulation and grading and staging were carried out separately. Hepatitis C virus RNA serum levels and viral genotype were compared in patients with or without liver iron accumulation. Alpha glutathione S-transferase serum levels were assessed in all patients. RESULTS: Overall, liver iron accumulation was mild and was present in 19 patients (36%). It was associated with male gender (P = 0.0358), and was reflected by high serum iron levels (P = 0.001) and high ferritin levels (P < 0.0001). Hepatitis C virus RNA levels and genotype were not associated with the presence of liver iron accumulation. In multivariate analysis, ferritin was the only variable significantly associated with liver iron accumulation (P < 0.0001). Grading was higher in patients with liver iron accumulation regardless of the site of iron deposition. Fibrosis was present in all patients with iron overload; these patients were more frequently cirrhotic. Moreover, patients with mesenchymal or mixed deposition had higher staging than patients with hepatocytic or no iron deposition. This feature was reflected by higher alpha-glutathione S-transferase levels. CONCLUSIONS: Liver iron accumulation is mild in chronic hepatitis C patients without HFE mutations and is mainly reflected by serum ferritin levels. Viral characteristics do not seem to play a role in iron deposition. Liver iron accumulation is associated with higher grading, advanced fibrosis and cirrhosis. Moreover, higher staging is associated with mesenchymal or mixed iron deposition. In these patients, higher alpha-glutathione S-transferase levels seem to reflect more complex damage.

Antiviral activity of amantadine in elderly patients with chronic hepatitis C.

BACKGROUND: Hepatitis C virus (HCV) infection is a worldwide problem of public health. Epidemiological studies have shown a significant higher prevalence of infection in the elderly. Amantadine is an antiviral agent active against the influenza A virus that has been used in cases of chronic hepatitis C. OBJECTIVES: To evaluate the antiviral activity and the safety of amantadine (200 mg daily for 6 months) in elderly patients with chronic hepatitis C. METHODS: The study group consisted of 23 consecutive patients over 65 years suffering from chronic hepatitis C. Aminotransferase (ALT) levels were tested at baseline, at 15 days and then monthly until the end of therapy. HCV genotype was determined at baseline. A quantitative HCV-RNA measurement was performed at baseline, at 15 days and at the 1st, 3rd and 6th month of treatment. RESULTS: 13 males and 10 females were enrolled (mean age 70.1 +/- 3.4 years; range: 65-75). The mean ALT levels did not change significantly during therapy except in 1 patient subsequently returned normal. The HCV-RNA remained detectable in all patients, but a significant difference in response was observed in patients infected by genotype 1b. CONCLUSIONS: Our results confirm the antiviral activity of amantadine against HCV, mainly for genotype 1b with initial high viral load. No consistent effects on aminotransferases were observed.

Probability of non-response during interferon therapy in patients with chronic hepatitis C.

BACKGROUND/AIMS: About 50% of patients with chronic hepatitis C do not respond to interferon therapy and this failure is expensive. The aim of this study was to identify possible predictive factors of biochemical non-response during interferon therapy among biochemical, virological (HCV genotype), histological (Knodell's score) and pharmacokinetic (monoethylglycinexylidide formation test) pre-treatment parameters. METHODOLOGY: Our study included 60 patients with chronic hepatitis C undergoing a course of Interferon therapy. Patients whose serum ALT levels were normal at the 3rd month of therapy and remained so until the end of treatment were regarded as responders. RESULTS: In univariate analysis, only the gamma-glutamyltransferase (gamma-GT) and the gamma-GT/alanine aminotranferase ratio were significantly higher in non-responder patients. Multivariate logistic analysis showed that high gamma-GT levels, high histological activity index, low monoethylglycinexylidide formation rate and viral genotype 1 were the best combination for the identification of non-responder patients (16.7% error rate). By adding alanine aminotranferase modification at the 1st month of therapy the probability error was reduced to 5%. CONCLUSIONS: These results show that the combination of biochemical, histological, virological and pharmacokinetic pre-treatment variables, associated with alanine aminotranferase modification at the 1st month of therapy, can predict non-response to interferon and allow therapeutic modifications.

Analytical and laboratory evaluation of a new fully-automated third generation enzyme immunoassay for the detection of antibodies to the hepatitis C virus.

An analytical and laboratory evaluation of a newly-developed fully-automated third generation ELISA for the detection of anti-HCV (Cobas Core Anti-HCV EIA, Roche) was undertaken. Coefficients of variation (CVs) calculated on positive control and serum samples ranged from 5.9 to 9.8% in the intra-assay precision test and from 3.9 to 11.3% in the inter-assay evaluation. With regard to the study of clinical laboratory performance, five groups of sera pre-screened with two third generation ELISA (Ortho HCV 3.0 ELISA; Innotest HCV Ab III) were assayed: anti-HCV negative samples (n = 932); anti-HCV positive samples (n = 449); difficult sera of different origin (n = 113); sera with discrepant results in the two ELISAs (n = 50); sera with an indeterminate result in one or more confirmatory test (n = 34). The overall concordance between the Roche anti-HCV EIA and the two reference assays was 97.5 and 97.8% for the Ortho and for the Innogenetics assays, respectively. Although it is not possible to provide absolute figures for clinical sensitivity and specificity, the results of the study on discrepant samples show that the Cobas Core Anti-HCV gives a number of negative results with positive or indeterminate confirmatory anti-HCV tests, which is intermediate between the Ortho and the Innogenetics assay. In contrast, only 5% Cobas Core Anti-HCV reactive sera are not positive or clear-cut single band reactive by supplemental assays. The results show that the new fully-automated third generation anti-HCV test is a valid alternative to other commercially available assays for screening of antibodies to the hepatitis C virus.

Anti-hepatitis C virus antibodies and hepatitis C virus viraemia in haemodialysis patients.

Sera from 82 haemodialysis patients were tested for anti-HCV, HCV-RNA, and HBsAg. Alanine aminotransferase (ALT) activity was monitored weekly for 2 months. Anti-HCV was positive in 31 patients (37.8%), showing different single-peptide patterns. HCV-RNA was detected in 26 anti-HCV-positive patients (84%) and also in two of 21 anti-HCV-negative patients. Twenty-seven (87%) of the 31 anti-HCV-positive patients had persistently normal ALT values; 22 of these patients were HCV-RNA positive. The four patients with elevated ALT values had HCV viraemia. HBsAg was positive in nine anti-HCV-negative patients. The close correlation between HCV viraemia and HCV status, independently of ALT values, requires that anti-HCV dialysis patients must be considered potentially infective and dialysed with reserved machines and/or in separate shifts.

Immunogenicity of a yeast-derived hepatitis B vaccine (Engerix-B) in healthy young adults.

In a study comparing the safety and immunogenicity of a recombinant DNA yeast-derived hepatitis B vaccine with that of a plasma-derived vaccine in young female adults, 50 subjects were vaccinated with the former and 29 with the latter vaccine according to a 0, 1, and 6 month vaccination schedule. Results indicated that the yeast-derived vaccine was safe and highly immunogenic. Two months after the second vaccine dose, 86% of subjects seroconverted, a rate which increased to 100%, 30 days after the booster dose. Moreover, anti-HBs geometric mean titres increased progressively after the first two doses and rose markedly to 1098 IU/l after the booster dose. Although similar rates of seroconversion were obtained with both vaccines, the anti-HBs GMT of the plasma-derived vaccine was higher (P less than 0.05) than that elicited by the yeast-derived vaccine.

Increasing prevalence of HTLV-III infection in intravenous drug users in Liguria, Italy.

Anti-HTLV III prevalence has been investigated in serum samples of 638 intravenous drug users collected over May 1981 - March 1985 and stored at -20 degrees C. Separately, in a prospective way, we have studied 68 IV drug abusers (53 Genoese and 15 of Sanremo area) of whom we have collected, at least, one serum specimen for each year, starting with 1981. We have also tested for anti-HTLV III presence serum samples of: 91 subjects of Hospital staff (Infectious Diseases Department and Laboratory workers); 32 workers in Therapeutic Communities for drug users and 24 family contacts of anti-HTLV III positive drug users. And then serum samples of two groups of general population collected for other seroepidemiological investigations in 1982 (256 subjects) and 1984 (538 subjects) were tested. No IV drug user was positive in 1981 whilst from 1982 up to 1984 there was a strong rising of the prevalence of anti-HTLV III positive subjects: 2, 22, and 39 per cent, respectively. The prevalence remained about 40% in the first months of 1985. The investigations carried out also show that HTLV III spread in Sanremo area slightly before than in Genoa and neighbourhood. No subject positive for anti-HTLV III has been detected among the Hospital staff and in workers of Therapeutic communities who have more probability to get in contact with infected subjects or their blood, as well as in the general population. A positive case has been discovered in a family contact (the wife of a positive for anti-HTLV III IV drug user). Some epidemiological and public health questions linked to the situation observed, at present, in Liguria, are discussed.

Prevalence of infections caused by AIDS and hepatitis B viruses in jailed people.

Prevalence of positive subjects to anti-HTLV III and HBV markers (HBsAg; anti-HBc; anti-HBs) has been studied both among jailed people and wardens of Sanremo Jail. Out of 92 subjects in custody, 11 were anti-HTLV III positive and 44 had acquired HBV infection markers (antigen and/or antibodies). One of the wardens resulted anti-HTLV III positive whilst 14 appeared to have been infected by HBV. All anti-HTLV III positive subjects, but the warden, were intravenous drug users. The study of prevalence was the first step of a perspective monitoring program in Ligurian Jails.

Passive-active immunoprophylaxis in newborns to HBsAg carrier mothers liguria--I.

Thirty neonates to HBsAg carrier mothers, two of whom were HBeAg positive, received passive-active immunoprophylaxis. Within 48 hours after birth and at the first month of life these newborns were given HBIG (0.5 ml/Kg) and at the third month of life they were given first vaccination (10 micrograms). "HB-Vax" (M.S.D.) has been employed and its schedule has been performed (2 degrees and 3 degrees injections respectively at 4th and 9th month of life). Blood specimens have been collected in infants at birth and at each HBIG and vaccine administration, two months after the 2nd vaccination, one and six months after the third one. All specimens have been tested for HBV markers: HBsAg, anti-HBs, anti-HBc, anti-HBe and anti-HBc IgM. The anti-HBs titration has been performed according to WHO International Reference anti-HBs Standard. ALT and AST levels were determined in all infants specimens. The newborns did not show clinical signs or lab tests expressing HBV infection during the follow-up period. The anti-HBs geometric mean titres have been the following: 125 mIU/ml at 1 month, 176.4 mIU/ml at 3 months, 64.4 mIU/ml at 4 months, 119 mIU/ml at 6 months, 321 mIU/ml at 9 months, 2580.8 mIU/ml at 10 months and 715.3 mIU/ml at 15 months of life. Finally, side effects were observed for no infants.

Quantitative evaluation of anti-HBs in sera--II.

We have investigated about the most correct way to express the anti-HBs titration in serum. The following linear regression has been elaborated on the basis of the results (converted into logarithms) obtained by 33 titrations of the International Reference Standard (the dilutions prepared were the following: 150, 100, 75, 37.5, 18.75, 9.375 mUI/ml) by as many kit lots AUSAB EIA (Abbott): y = 0.8590732x - 1.6936435. The correlation between the couples: optical density log. mUI/ml has been highly significative (r = 0.991091; P less than 0.01). In order to limit the variability effects in the same laboratory, the use of the correction factor "F" is suggested. The variation of the optical density corrected by the "F" variation is illustrated. A linear regression on the Author's data has been elaborated by the inverse application of the Hollinger formula. The adaptability of this linear regression has been tested on 196 dilutions out of the 33 standard tests studied. The average values obtained, the 95% confidence limits and the mean values +/- 2s limits are compared with the same values obtained by applying the Hollinger original formula.

Quantitative evaluation of anti-HBs in sera.

The Authors carried out a methodological study about the way of expressing in mIU/ml the antibody to HBV surface antigen (anti-HBs) titer. The results showed that Hollinger's formula seems to provide a good accuracy when enzyme immunoassay is employed (AUSAB EIA, Abbott). Moreover, moderate variations in the commercial anti-HBs kits studied (AUSAB, AUSAB EIA, Abbott) have been verified. Considering that and the progress of the calibration curve of the WHO Reference International Standard, it is also advised to include always in the test a calibration curve of the standard serum when a higher precision is required.