Viral, metabolic, and genetic causes of liver disease were exclud

Viral, metabolic, and genetic causes of liver disease were excluded by appropriate investigations, all patients being negative for anti–hepatitis C virus, hepatitis B surface

antigen, Epstein-Barr virus, and cytomegalovirus serological markers of active infection. From 9 of 16 [A] patients, blood was obtained before treatment; the remaining 7 [A] patients were studied at relapse during immunosuppression tapering (3 were on 4 mg of methylprednisolone daily and 4 were on 2-4 mg of methylprednisolone and 50 mg of azathioprine daily). The 31 [R] patients had normal alanine aminotransferase (ALT) and gamma-globulin levels for a median of 40 months (range = 8-120 months); 21 were on 2 to 4 mg of methylprednisolone daily, 3 were on 50 mg of azathioprine daily, and 7 were on 2 to 4 mg of methylprednisolone and 50 to 100 mg of azathioprine daily. Everolimus solubility dmso The median duration of immunosuppression was 42 months (range = 12-237 months). Liver biopsy showed histological features of interface hepatitis in all 38 patients at diagnosis. In the

group of [A] patients, ANAs were present in 12 patients, SMAs were present in 6, soluble liver antigen was present in 1, and anti-mitochondrial antibodies were present in 1 (ANAs and SMAs find more co-occurred in 3 individuals). At diagnosis, all 31 [R] patients tested positive for autoantibodies (ANAs and SMAs were detected in 19 and 21 subjects, respectively, and co-occurred in 15), whereas at the time of study, 7 (28%) had lost all autoreactivity (ANAs and/or SMAs were still present in 22 subjects and co-occurred in 4). At diagnosis, all patients fulfilled the diagnostic criteria of the International 上海皓元 Autoimmune Hepatitis Group.5 Clinical and laboratory features are summarized in Table 1. All 16 [A] patients had high aminotransferase, gamma-glutamyl transpeptidase (GGT), and bilirubin levels, increased international normalized ratios (INRs), high gamma-globulin and immunoglobulin G (IgG) levels, and seropositivity for autoantibodies. All [R] patients had normal biochemical tests, but autoantibodies were still detectable in half. Sera

were tested for non–organ-specific autoantibodies by indirect immunofluorescence on cryostatic sections of rat liver, kidney, and stomach specimens at an initial serum dilution of 1:40.33 Anti–soluble liver antigen was detected by enzyme-linked immunosorbent assay according to the manufacturer’s instructions (Euroimmun, Lubeck, Germany). Peripheral blood mononuclear cells (PBMCs) were prepared from 20 mL of peripheral blood with preservative-free heparin (10 U/mL), diluted 1:1 with Roswell Park Memorial Institute 1640 (RPMI-1640) medium (Invitrogen Life Technologies, Paisley, United Kingdom), and separated with Ficoll-Hypaque (Amersham Pharmacia Biotech, Ltd., Little Chalfont, United Kingdom). PBMCs were collected and washed twice with RPMI-1640. Viability, determined by trypan blue exclusion, always exceeded 98%.

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