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HGV in patients with autoimmune hepatitis: anti-E2 antibodies and HGV-RNA in serum
226 IMMUNOLOGY, AUTOIMMUNE LIVER DISEASES pizq
1 co9/003
HGV IN PATIENTS WITH AUTOIMMUNE HEPATITIS: ANTI-E2 ANTIBODIES AND HGV-RNA IN SERUM
DETECTION OF HUMAN Mx PROTEIN M PARAFFIN-WAX EMBEDDED LIVER TISSUE
B. Tribl. M. Sch(iniger-Hekele. D. Petermann. S. Bakos. E. Penner, C. Miiller. Department of Internal Medicine IV, University of Vienna, Austria. Aims: We studied the prevalence of HGV-RNA in serum by reverse PCR and determined antibodies against the envelope protein E2 of HGV by ELISA in 83 patients with welldefined autoimmune hepatitis (AM). Methods: HGV-RNA was detected by reverse transscription PCR with nested primers from the non-structural region 3. Sequence data were obtained by an automatic sequenzer. Anti-E2 antibody, a marker of past infection, was measured by ELISA. Results: HGV-RNA was found in 11 of 83 patients (13%) as compared with 2% of healthy controls (p=O.O12). Anti-E2 was observed in 15 (18%) patients which was not different from healthy controls (12 out of 93; ns.). Only 1 patient had both HGV-RNA and anti-E2 present in serum at the same time. HGV-RNA was found in 8% of type I AIH, but in 23% and 14% of type II and type III AIH, respectively. Conversely, 26% of type I, but only 12% and 7% of type II and type III AlH had anti-E2. Exposure to HGV (HGV-RNA+ or anti-E2+) was similar in all three types (34%, 35%, 2 1%)).Sequence analysis of HGV in the 11 positive patients revealed infection with only one single genotype of HGV (type 2b). Conclusions: Patients with AIH have an increased risk for HGV infection. Patients with type I AIH are more likely to show anti-E2 as a marker of past HGV infection. Only a single genotype of HGVRNA was present in our population of patients with AIH.
1
M. Vassilev, KAntonov, I. Michailov*, Z. Krastev Clinic of Gastroenterology, *Department of Pathology, Medical University, Sofia, Bulgaria([email protected],bg) Interferons modulate a number of biological functions including cell growth and differentiation, the immune response, and virus replication. The MxA gene expression is regulated exclusively by type I interferons and is a good marker for detecting minute quantities of biologically active type I IFN. An immunohistochemical method for the detection of human Mx proteins in par&n-wax embedded liver tissue was developed. Microwave pretreatment for epitope retrieval in citrate buffer and anti human Mx moAb kindly provided by Dr. Hochkeppel, Novartis pharma Research Base1 were used.Twenty four liver biopsies from patients (lOE/14m; 40.58+/-10.85 yr.) with chronic viral liver diseases (HBV - 6; HCV - 8; HBV+HCV -1) and from patients without virus-related changes in the liver histology(g) were studied. Staining for Mx protein was found in all specimens studied and in all cell types, although of different intensity and with different pattern. In general, zones of diffuse high intensity cytoplasmic staining for Mx protein was seen in viral liver diseases, whereas, low intensity cytoplasmic staining and foci of discrete granules in hepatocytes with or without staining of sinusoidal cells were characteristic for normal liver. Mx positive nonparenhymal cells were seen in the inflammatory lesions. These findings show that Mx protein can be visualized in paraffin-wax embedded liver tissue and that there are different patterns of Mx protein expression - both at cellular and tissue level.
1 cog/o04 1 INTERSTITIAL LUNG FIBROSIS IN PATIENTS WITH HEPATITIS C VIRUS INFECTION. C. Feni. L. La Civita, P. Fazzi*. M. Monti’. G. Longombardo, G Careccia”, S. Solfanelli*. G. Porciello. G. Pasero. P. Gentilini’. A L Zirmeao”. Department of Internal Medicine and *Respiratory Disease Unit, University of Pisa; “Institute of Internal Medicine, University of Florence, Florence, Italy. Hepatitis C virus (HCV) chronic infection is a multifaceted disorder characterised by hepatic and extra-hepatic immune-lymphoproliferative mixed autoimmune hepatitis, cryoglobulinemia, disorders; namely, thyroiditis, glomemlonqkitits, and B-cell lymphomas. Moreover, HCV has been correlated with ‘idiopathic’ lung fibrosis. Here, a cohort of 10 pts (M/F = 416, mean age 6OilOSD years) with lung fibrosis belonging to our series of 300 HCV-infected pts is described. Lung fibrosis appeared medially Sk4SD years after the clinical onset of chronic hepatitis which was clinically evident in S/lOpts. Moreover, lung involvement was often associated with the following disorders: arthritis (5 pts); sicca syndrome (1 pt); severe polymyositis and cranial neuropathy (1 pt); circulating mixed cryoglobulins and/or auto-antibodies (8 pts). Lung involvement was suspected on the basis of clinical symptoms (dyspnoea, cough, clubbing) and/or pulmonary abnormalities at routine chest X-ray. Interstitial lung fibrosis was quantified (moderate in 4, severe in 6 pts) by high resolution computed tomography. The presence of active lung involvement was suggested by parenchymal uptake of 67Ga (10110 pts) and increased percentages of neutropbils and/or lymphocytes (5/5 pts) at bronchoalveolar lavage. A significant reduction of Dlco and spirometric abnormalities were observed in lO/lO and 500 pts were respectively. In all cases, serum anti-HCV and HCV-RNA demonstrated; HCV genomic sequences were also detected in peripheral lymphocytes (4/4pts) and in one case in lung biopsy specimens. A desquamative interstitial pneumonia was demonstrated in 2 pts by lung biopsy. These data further support the hypothesis that HCV can trigger a constellation of immune-mediated disorders including the interstitial lung fibrosis.
HEPATIC AUTOANTIGENS IN THE AUTOIMMUNE POLYGIANDULAR SYNDROME TYPE 1 AND AUTOIMMUNE HEPATITIS TYPE 2 P. Obermaver-Straubl, S. Braunl, S. Lo!aes, M.G. Clementel, B. Liittiq’, B. Grams’, J. Pemeentupaz and M.P. Mannsl l Dept. of Gastroenterology, Hannover Medical School, Germany zChildren’s Hospital, University of Helsinki, Finnland. APS-1 is a rare autoimmune disease characterized by the progressive manifestation of a variety of disease components. 7168 APSI patients developed hepatitis. Since CYP2D6 is the major autoantigen in autoimmune hepatitis type 2 (AIH-2) all 68 APS-I sera ware tested for CYP2D6 autoantibodies. However no signals were detected in Western Blots with recombinant CYP2D6. Tests with preparations of microsomes containing eight different recombinant CYPs, namely CYPIAI, CYPlAL, CYP2A6, CYP2B6, CYP2C9, CYP2C18, CYP2E1, CYP3Al revealed CYPlA2 and CYP2A6 as major hepatic autoantigens in APSI. CYPfA2 was recognized by 4/68 APSI sera, CYP2A6 by 9/68 sera. Minor hepatfc autoantigens are CYPIAI and CYP2B6, which were recognized by one exceptional serum. When the results were compared with patient data all 4 patients with anti-CYPIAP were affected by hepatitis, however only 2/9 patients sera, which recognize CYP2A6. This result indicates that antiCYPlA2 autoantibodies are specific for hepatitis in APSI and that anfCYP2A6 autoantibodies may be linked to another disease component in APSI. When 40 sera from patients with AIH-2 were tested for antiCYPiA2 in Western Blots, none of the sera revealed binding activity. These results demonstrate that hepatic autoantibodies in APSI differ from autoantibodies in idiopathic autoimmune hepatitis. Therefore in children affected by hepatitis of autoimmune ethiology always the identity of the target protein should be determined in order to distinguish between idiopathic autoimmune hepatitis and hepatitis as disease component in APSI, which may be followed by other serious autoimmune diseases.
1 co9/003
HGV IN PATIENTS WITH AUTOIMMUNE HEPATITIS: ANTI-E2 ANTIBODIES AND HGV-RNA IN SERUM
DETECTION OF HUMAN Mx PROTEIN M PARAFFIN-WAX EMBEDDED LIVER TISSUE
B. Tribl. M. Sch(iniger-Hekele. D. Petermann. S. Bakos. E. Penner, C. Miiller. Department of Internal Medicine IV, University of Vienna, Austria. Aims: We studied the prevalence of HGV-RNA in serum by reverse PCR and determined antibodies against the envelope protein E2 of HGV by ELISA in 83 patients with welldefined autoimmune hepatitis (AM). Methods: HGV-RNA was detected by reverse transscription PCR with nested primers from the non-structural region 3. Sequence data were obtained by an automatic sequenzer. Anti-E2 antibody, a marker of past infection, was measured by ELISA. Results: HGV-RNA was found in 11 of 83 patients (13%) as compared with 2% of healthy controls (p=O.O12). Anti-E2 was observed in 15 (18%) patients which was not different from healthy controls (12 out of 93; ns.). Only 1 patient had both HGV-RNA and anti-E2 present in serum at the same time. HGV-RNA was found in 8% of type I AIH, but in 23% and 14% of type II and type III AIH, respectively. Conversely, 26% of type I, but only 12% and 7% of type II and type III AlH had anti-E2. Exposure to HGV (HGV-RNA+ or anti-E2+) was similar in all three types (34%, 35%, 2 1%)).Sequence analysis of HGV in the 11 positive patients revealed infection with only one single genotype of HGV (type 2b). Conclusions: Patients with AIH have an increased risk for HGV infection. Patients with type I AIH are more likely to show anti-E2 as a marker of past HGV infection. Only a single genotype of HGVRNA was present in our population of patients with AIH.
1
M. Vassilev, KAntonov, I. Michailov*, Z. Krastev Clinic of Gastroenterology, *Department of Pathology, Medical University, Sofia, Bulgaria([email protected],bg) Interferons modulate a number of biological functions including cell growth and differentiation, the immune response, and virus replication. The MxA gene expression is regulated exclusively by type I interferons and is a good marker for detecting minute quantities of biologically active type I IFN. An immunohistochemical method for the detection of human Mx proteins in par&n-wax embedded liver tissue was developed. Microwave pretreatment for epitope retrieval in citrate buffer and anti human Mx moAb kindly provided by Dr. Hochkeppel, Novartis pharma Research Base1 were used.Twenty four liver biopsies from patients (lOE/14m; 40.58+/-10.85 yr.) with chronic viral liver diseases (HBV - 6; HCV - 8; HBV+HCV -1) and from patients without virus-related changes in the liver histology(g) were studied. Staining for Mx protein was found in all specimens studied and in all cell types, although of different intensity and with different pattern. In general, zones of diffuse high intensity cytoplasmic staining for Mx protein was seen in viral liver diseases, whereas, low intensity cytoplasmic staining and foci of discrete granules in hepatocytes with or without staining of sinusoidal cells were characteristic for normal liver. Mx positive nonparenhymal cells were seen in the inflammatory lesions. These findings show that Mx protein can be visualized in paraffin-wax embedded liver tissue and that there are different patterns of Mx protein expression - both at cellular and tissue level.
1 cog/o04 1 INTERSTITIAL LUNG FIBROSIS IN PATIENTS WITH HEPATITIS C VIRUS INFECTION. C. Feni. L. La Civita, P. Fazzi*. M. Monti’. G. Longombardo, G Careccia”, S. Solfanelli*. G. Porciello. G. Pasero. P. Gentilini’. A L Zirmeao”. Department of Internal Medicine and *Respiratory Disease Unit, University of Pisa; “Institute of Internal Medicine, University of Florence, Florence, Italy. Hepatitis C virus (HCV) chronic infection is a multifaceted disorder characterised by hepatic and extra-hepatic immune-lymphoproliferative mixed autoimmune hepatitis, cryoglobulinemia, disorders; namely, thyroiditis, glomemlonqkitits, and B-cell lymphomas. Moreover, HCV has been correlated with ‘idiopathic’ lung fibrosis. Here, a cohort of 10 pts (M/F = 416, mean age 6OilOSD years) with lung fibrosis belonging to our series of 300 HCV-infected pts is described. Lung fibrosis appeared medially Sk4SD years after the clinical onset of chronic hepatitis which was clinically evident in S/lOpts. Moreover, lung involvement was often associated with the following disorders: arthritis (5 pts); sicca syndrome (1 pt); severe polymyositis and cranial neuropathy (1 pt); circulating mixed cryoglobulins and/or auto-antibodies (8 pts). Lung involvement was suspected on the basis of clinical symptoms (dyspnoea, cough, clubbing) and/or pulmonary abnormalities at routine chest X-ray. Interstitial lung fibrosis was quantified (moderate in 4, severe in 6 pts) by high resolution computed tomography. The presence of active lung involvement was suggested by parenchymal uptake of 67Ga (10110 pts) and increased percentages of neutropbils and/or lymphocytes (5/5 pts) at bronchoalveolar lavage. A significant reduction of Dlco and spirometric abnormalities were observed in lO/lO and 500 pts were respectively. In all cases, serum anti-HCV and HCV-RNA demonstrated; HCV genomic sequences were also detected in peripheral lymphocytes (4/4pts) and in one case in lung biopsy specimens. A desquamative interstitial pneumonia was demonstrated in 2 pts by lung biopsy. These data further support the hypothesis that HCV can trigger a constellation of immune-mediated disorders including the interstitial lung fibrosis.
HEPATIC AUTOANTIGENS IN THE AUTOIMMUNE POLYGIANDULAR SYNDROME TYPE 1 AND AUTOIMMUNE HEPATITIS TYPE 2 P. Obermaver-Straubl, S. Braunl, S. Lo!aes, M.G. Clementel, B. Liittiq’, B. Grams’, J. Pemeentupaz and M.P. Mannsl l Dept. of Gastroenterology, Hannover Medical School, Germany zChildren’s Hospital, University of Helsinki, Finnland. APS-1 is a rare autoimmune disease characterized by the progressive manifestation of a variety of disease components. 7168 APSI patients developed hepatitis. Since CYP2D6 is the major autoantigen in autoimmune hepatitis type 2 (AIH-2) all 68 APS-I sera ware tested for CYP2D6 autoantibodies. However no signals were detected in Western Blots with recombinant CYP2D6. Tests with preparations of microsomes containing eight different recombinant CYPs, namely CYPIAI, CYPlAL, CYP2A6, CYP2B6, CYP2C9, CYP2C18, CYP2E1, CYP3Al revealed CYPlA2 and CYP2A6 as major hepatic autoantigens in APSI. CYPfA2 was recognized by 4/68 APSI sera, CYP2A6 by 9/68 sera. Minor hepatfc autoantigens are CYPIAI and CYP2B6, which were recognized by one exceptional serum. When the results were compared with patient data all 4 patients with anti-CYPIAP were affected by hepatitis, however only 2/9 patients sera, which recognize CYP2A6. This result indicates that antiCYPlA2 autoantibodies are specific for hepatitis in APSI and that anfCYP2A6 autoantibodies may be linked to another disease component in APSI. When 40 sera from patients with AIH-2 were tested for antiCYPiA2 in Western Blots, none of the sera revealed binding activity. These results demonstrate that hepatic autoantibodies in APSI differ from autoantibodies in idiopathic autoimmune hepatitis. Therefore in children affected by hepatitis of autoimmune ethiology always the identity of the target protein should be determined in order to distinguish between idiopathic autoimmune hepatitis and hepatitis as disease component in APSI, which may be followed by other serious autoimmune diseases.