Type 1 autoimmune hepatitis: Clinical course and outcome in an Italian multicentre study

Abstracts / Digestive and Liver Disease 38 (2006) A75–A85

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and HeLa nucleolar extract were used as substrates in Western blot (WB) analysis. Cell extracts of [35 S]methionine-labelled HeLa cells were used as antigens in immunoprecipitation (IP). Different phases synchronised cells were also used to confirm antigen expression in IIF, WB, IP. Results. Five out of 105 (4.8%) PBC sera showed a nucleolar cell cycle related staining. None of the control sera showed this unique IIF pattern. Interestingly, one out of these five PBC sera was AMA negative. In cell synchronisation experiments, double IIF staining with a rabbit anti-PCNA or anti-CENP-F serum with these three human sera showed an identical nucleolar staining pattern only in the nucleoli during the G2 phase. Human sera did not show reactivities in WB and IP. Conclusions. This study describes a new autoantibody reactivity to a novel nucleolar antigen found in PBC patients. This reactivity seems highly specific for PBC and it could help establishing the correct diagnosis in AMA negative patients with PBC. Further studies will be necessary to define the nature of this novel autoantigen.

atic autoimmune disease (13.5%): SSc in two patients (one HCV+ and one cryptogenic), Graves disease in one (HCV+), Rheumatoid Arthritis in one (HCV+) and coeliac disease in one (AIH). Five patients (13.5%) had an extra hepatic neoplasm (three had had a breast cancer preceding ANA positivity, one rectal carcinoma and one bladder carcinoma 2 year after ANA positivity). Seventeen patients had sequential ANA tests available and 13 (76%) changed nucleolar pattern during follow-up. Eight were HCV+: three (37%) developed ANA-N reactivity during an hepatitic flare, two loose ANAN after virological response to interferon and two returned ANA-N+ at the time of virological relapse. 1 HBV+ patient developed ANA-N during a flare of viral replication. All three AIH loose nucleolar pattern during immuno-suppressive therapy but one became positive again at the time of hepatitis reactivation. Conclusions. ANA-nucleolar pattern seems to be associated with autoimmune manifestations, with development of extrahepatic neoplasms and in viral hepatitis it seems to correlate with disease flares.

doi:10.1016/j.dld.2006.06.019

doi:10.1016/j.dld.2006.06.020

SERUM NUCLEAR AUTOANTIBODIES WITH NUCLEOLAR PATTERN IN CHRONIC LIVER DISEASES

TYPE 1 AUTOIMMUNE HEPATITIS: CLINICAL COURSE AND OUTCOME IN AN ITALIAN MULTICENTRE STUDY

V. Monti, E. Arosio, M.F. Donato, M. Berra, A. Aghemo, M. Colombo

E. Rosa Rizzotto a , M. Durazzo b , G. Niro c , S. Antoniazzi a , F. Ferrara a , I. Carderi a , V. Baldo d , A. Premoli c , F. Olivero c , E. Morello c , A. Floreani a

Department of Gastroenterology, IRCCS Fondazione Policlinico Mangiagalli e Regina Elena, Milan, Italy Background. Circulating nuclear autoantibodies with a nucleolar pattern (ANA-N) have been associated with systemic sclerosis (SSc) or neoplasms including hepatocellular carcinoma. Aim. To study the clinical impact of ANA-nucleolar pattern in patients with chronic liver diseases. Material and methods. Retrospective analysis of patients registered as having a chronic liver disease and ANA test positive performed between 2001 and 2004, on Hep-2 cells substrate by immuno-florescence. Serial ANA tests were available in a subgroup of cases. All ANA positive sera were tested for antigen specificity (ENA, dsDNA, centromerus). Patients features, presence of extrahepatic autoimmune disease and neoplasms prevalence were analysed. Results. Thirty-seven out of 811 serum ANA+ patients showed an ANA-N pattern (4.6%). ANA titres ranged between 1:80 and >1:320. Sixty-two percent were female, with a mean age of 52 ± 13year. Twenty-six had HCV+ (46% genotype 1), four HBV+, three were Autoimmune Hepatitis, two Primary Biliary Cirrhosis and two Cryptogenic Hepatitis. Antigenic specificity was: anti Scl70 in two (1 HCV+ and 1 cryptogenic), anti centromeric in one (HCV+) and anti dsDNA+ in three (two AIH and one PBC). Ten (27%) patients had well compensated cirrhosis. Five patients had extra hep-

a

Department of Surgical and Gastroenterological Sciences, University of Padova, Padua, Italy b Department of Medicine, Ospedale Molinette, Turin, Italy c IRCCS, S. Giovanni Rotondo, Italy d Department of Hygiene and Public Health, University of Padova, Padua, Italy Diagnosis of AIH is based on a combination of clinical, biochemical and histological features together with careful exclusion of other causes of liver disease. Many reports of the condition were written in the ‘pre Hepatitis C era’ or were based on data from a limited number of centres. Data on the natural history are still lacking. Aim. To evaluate the clinical presentation and the natural history of type I AIH. Methods. Seventy-three consecutive patients (63 females, 10 males) with a regular follow-up of at least 2 years were prospectively included in the study. The mean followup was 91 + 61 months. The International Autoimmune Hepatitis Group (IAHG) revised scoring was applied to each patient. Treatment included immunosuppression (prednisolone associated with Azathioprine) with a tailored maintenance dosage. Results. Patients with ‘acute’ onset at presentation (N. 27) were significantly older (56 + 20 years) than patients with ‘chronic’ onset (45.5 ± 15 years, p < 0.05) and had signifi-

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cantly higher serum levels of AST/ALT, GGT, bilirubin; PT was significantly lower in the said group compared to patients with AIH with ‘chronic’ onset. The total number of pregnancies was 94: 88 (93.6%) before the onset of the disease, and 6 (6.4%) after the diagnosis of AIH, during the pharmacological remission. Among these, 2 patients experienced discharge, and the remaining had a normal vaginal delivery. At overall no alteration of the fertile life was noted. Particularly the mean age of menarche was 12.8 ± 1.7 years, and the mean age of menopause was 50.5 ± 3.8 years. The major events during the follow-up included: oesophageal varices (N. 9), and ascitis (N. 4), whereas 60 patients remained in remission while receiving immunoppression. None of the patients died during follow-up, but 7 patients were transplanted. The cumulative transplant free probability of survival was 80% at 280 months. Conclusions. Survival in AIH is apparently good when immunosuppression is protracted for ever.

observation to genotype 1 patients. No difference was found between Cryo+ and Cryo−. When compared to the other patients, Cryo++ were significantly older, (p < 0.0001), more frequently females (p < 0.002), with lower IgG (p < 0.00004), higher IgM (p < 0.005), and with similar liver histology. By stepwise logistic regression analysis, age and number of HCV-Ag positive cells were independently significantly associated with Cryo++. Conclusions. In HCV-related CS a higher liver HCV-Ag expression associated to a longer disease duration may represent the basis for the activation of the mechanism that lead to vasculitis. In fact we can speculate that the availability of HCV-Ag and the formation and persistence of immunocomplexes within the circulation represents the basis for the cryoprecipitation and the activation of the inflammatory response responsible for the vasculitis. Whether the increased amount of HCV in the liver of CS patients is secondary to a condition of immunotolerance of the host remains speculative.

doi:10.1016/j.dld.2006.06.021 doi:10.1016/j.dld.2006.06.022 HIGH LIVER HCV ANTIGENS (HCV-AG) EXPRESSION IN HCV-RELATED CRYOGLOBULINAEMIC SYNDROME (CS) A. Grassi, G. Ballardini, M. Susca, C. Quarneti, G. Farina, G.P. Bianchi, V. Cipriano, F.B. Bianchi, M. Lenzi Dipartimento di Medicina Interna, Cardioangiologia, Epatologia, Policlinico S.Orsola-Malpighi, Universit`a di Bologna, Bologna, Italy Background. HCV has been recognised as the major aetiologic factor of CS, possibly through B-cell clonal proliferation sustained by HCV-Ag. The aim of this study was to evaluate the expression of HCV-Ag in liver samples of patients infected with HCV with (Cryo++) and without (Cryo−) CS. Material and methods. The number of hepatocytes positive for HCV-Ag were evaluated in 213 consecutive HCV+ patients with an available frozen liver biopsy. HCV-Ag were detected using an immunoperoxidase technique. The number of HCV infected hepatocytes was scored as follows: 0 = negative, 1 = <5%, 2 = 5–20%, 3 = >20% positive hepatocytes. The intensity of staining was scored from 1 to 3 (faint, medium, strong). Results. Circulating cryoglobulins were found in 77/213 patients: 55 without (Cryo+) and 22 with CS (Cryo++). HCV-Ag were more frequently detected in Cryo++ (86%) with respect to Cryo+ (65%) and Cryo− (61%) (p = NS and p < 0.04, respectively). Mean scores of HCV staining were significantly higher in Cryo++ (2.0) versus Cryo+ (1.3) and Cryo− (1.2) (p < 0.03 and p < 0.002, respectively). Similarly, mean intensity of staining in positive cases was higher in Cryo++ (2.1) with respect to Cryo+ (1.7) and Cryo− (1.5) (p = 0.05 and p < 0.001, respectively). This was maintained limiting the

NATURAL COURSE AND INCIDENCE OF NONORGAN SPECIFIC AUTOANTIBODIES (NOSA) AND HCV INFECTION IN THE GENERAL POPULATION: A NESTED CASE-CONTROL STUDY OF THE DIONYSOS COHORT M. Guidi a , A. Granito a , M. Lenzi a , L. Miglioli b , L.S. Croc`e c , P. Muratori a , A. Castiglione c , L. Muratori a , F.B. Bianchi a , G. Bedogni b,c , C. Tiribelli c , S. Bellentani b a

Departiment of Internal Medicine, Cardioangiology, Hepatology, University of Bologna, Bologna, Italy b Centro Studi Nutrizione e Fegato, Ospedale di Carpi (Modena), Italy c Centro Studi Fegato, Science Park, Basovizza (Trieste), Italy This study aims to explore the natural course and the crude incidence of non-organ specific autoantibodies (NOSA), and the presence and severity of chronic liver disease (CLD) in the HCV infected subjects of the Dionysos cohort. Methods. After a median period of 8.5 years from the first (1992) screening, 122 (69 M and 53 F) out of the 161 HCVRNA positive subjects (HCV+) were screened (compliance = 76%), and paired-matched for sex and age with 122 normal subjects of the same cohort (controls = C). Sera were tested for the presence of NOSA (antinuclear antibody-ANA, anti-smooth muscle antibody-SMA and anti-liver/kidney microsomes type 1-LKM1) by indirect immunofluorescence (IFL) at a 1:40 serum dilution. The annual crude mortality rate of the entire cohort was also registered. Results. 24/122 patients (20%) were treated with one or more courses of IFN and only 4 (all genotype non-1) are longterm responders. Antiviral treatment was not associated with variation in the autoantibody profile. The overall prevalence