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HLA-H mutations in patients with chronic hepatitis C
144
SWEDISH PATIENTS WITH HEMOCHROMATOSIS (HH) HAVE LOW LEVELS OF IRON AND A LOW FREQUENCY OF CIRRHOSIS
K. Hagen’, R. Befrits’, E. M. P. Cardoso’, P. Sti*, R. Hultcrantz’. ‘Department of Gastroenterology and Hepatology, Karolinska Hospital, Stockholm, *Department of Gastroenterology and Hepatology, Huddinge Univ. Hospital, Huddinge, Sweden. In recent reports, the frequency of liver cirrhosis in patients with HH is 60-70%. Liver cirrhosis is caused bv hick liver iron stores. due to late diagnosis or a more intense exbresiion of the disease. Cirrhosis may also he caused by coexisting factors, such as hepatitis B or C virus infection, or alcohol overconsumption. We studies these factors in a Swedish HH oatient erouo. The group consisted of 76 patients (56 men &d 17 women) with HH, diagnosed between 1980 and 1996 in Stockholm. All patients were homozygous for the C282Y mutation. 9 patients (11.8%) had cirrhosis, 23 (30.3%) had fibrosis and 44 (57.9%) had no fibrosis. The mean iron removed was 6.71 f 4.1 g (0.9-23.4 g), 7.2 f 4.3 g (1.4-23.4 g) for men and 5.0 f 2.9 g (0.9-14.0 g) for women. 30 patients (39.5%) had less than 5 g of removable iron. AU patients ;vith cirrhosis had iron stores exceeding 7 g. Of the patients without fibrosis, only 7 patients (15.9%) had iron stores exceeding 7 g. Iron stores were not age correlated. The mean aee at diagnosis was 47.4 f 13.5 yea& (20-73 years), 46.9 f 13.3 years (20-73 years) for men and 50.2 f 13.9 years (25-72 years) for women. 2 patients (males) were infected with hepatitis B virus, no patient with hepatitis C virus. 5 of the patients (3 men and 2 women) consumed more than 80 g alcohol/day. Conclusion: Swedish HH patients have a low frequency of cmhosls. rfijljs is not due to younger age at diagnosis, but is due to lower iron stores and a lower frequency of coexisting liver damaging factors.
REDOX GENE THERAPY OF LIVER ISCHEMIA/ REPERFUSION INJURY REDUCES AP-1 AND NF-KB ACTIVATION
Ralf M. Zwacka2, Yulong Zhanalt, Jeff Halldorsonl2, Lorita Dudus 12,and John F. Enaelhardt 1 1 University
of Iowa, Department
of Anatomy
and Cell
2 University of Edinburgh, Dept. Biology, Iowa City, Iowa. of Surgery, MRC Human Genetics Unit, Western General Hospital, Edinburgh. Liver transplantation is the only therapeutic strategy for numerous inherited and acquired diseases. The formation of reactive oxygen species (ROS) following ischemia/reperfusion (l/R) have been hypothesized to play a major role in transplant associated hepatocellular injury and ultimately graft rejection. Here, we describe the application of mitochondrial manganese superoxide dismutase (MnSOD) gene transfer to the liver as a therapeutic strategy for I/R injury in the liver of mice. Adenoviral-mediated overexpression of MnSOD in the liver of nulnu mice prior to lobar l/R achieved a 50% inhibition of acute liver damage. Associated activation of both NF-KB and AP-1 was also attenuated in animals expressing recombinant MnSOD but not in those expressing an irrelevant transgene pgalactosidase. These findings suggest that changes in the cellular redox state regulate the activity of transcription factors that are involved in the acute cellular response to I/R damage. These studies provide the foundation for redox mediated gene therapies for I/R injury which has been associated with acute rejection in autologous liver transplantation.
Topi E, k I. Tdija A,.I&ik7,,cekadaS. Antoljak N, .$tefanoviCM Clinical timte of Chem&ry and Department of pediatrics, !&he milosrdnice Univaity Hospital and S&ool of Medicine University of a=b, Kantrids Chikhen’s Hopital Rijcka University Hospital &&a, Croatia Deficiency alleles of al-anti~psin (alAT) are associated with low semm levels of alAT and can be either linkcd to liver disease and/or to early w emphysema. The most common deficiency alleles are PiS and Z. PiZ genotype is charade&cd hy G+A substition in exon 5 and change of Glu to Lys at aminoacid position 342. PiS genotype is ckuacterkd by substitotion of base A-T in exon 3 and aminmcid change of Glu to Val at p&ion 264. The aim of this study was to investigate the conco&nce between phm and genotype of al antimine polytnorpbic alleles Pi S snd Z using mutant primers in FCR-RFLp analysis. In four families (mother, father and child) phenotypes wa dctemkcd by iswlearic focusing (Pbannacia, PAG-plate pH 4D5.0) and gewypes by KR-RFLP, using mutant pimers (MWG-Biotech) for FCR, Taq I @&a) for amplicon dig&on, and 3% Meta Phor agamsc @MC Bio plodud) for electmphotetic sepamtion of restiction 6agtnerl.s. The serum alAT concentration was measured by RLD methoci.FamilyAwas~MZandMZphenotypeandgenotypeforfather motbcr and chid. Each member of this family had alAT cotwntmtion in normal ~ge,FamilyBwas~~22phenotype~genotypefor~,mmotherand child @vely. only the child in this family has madwlly decreased alAT concentration. In family C. disnepancy between @cnw-gcndypc was found: a boyptwentedZZphnwtypebuthJZgenotype.Hismc#lierwasMMandfather MZ gehotypc and phenotype. Tk cowenhation of alAT in each member of family C was in normal range. Results of family C reassessment indkatai that the discrepancy was not due to sample miskd@ or technical difficulties but a hue discordance,FamilyDwas~MZand22phenotypeandgenotypeforfather, mother and child rcspe&ely. The alAT CQncentration in this family was markcdly derreased only in child. The diw between genotype aal phenotype should he thomughly studied to identify the possible caw oP changes wzcuning along the gene to pmtein cycle.
HLA-H MUTATIONS IN PATDENTSWITH CHRONIC HEPATITIS C.
P. Ferenci. Ch. Datz. L. Kazemi-Shirazi. W. Ha&l. Th. MaierDobersberger. C.Polli. K. Kaserer. Ch. Osterreicher, P. Steindl, F. Wrba, E. Penner. Vienna, Linz, and Salzburg, Austria To determine a role of genetic hemochromatosis for iron accumulation in chronic hepatitis C (CH-C) the mutations of the HLA-H gene (Cys282Tyr, His63Asp) were evaluated in 98 (70 male/28 female) patients with biopsy-confirmed CH-C. Transferrin saturation (Tf?/o) was < 45% in 64, and >45 (range: 46-89) in 34. Hepatic iron content was measured by atom absorption spectroscopy and the hepatic iron index (I-III) was calculated. Mutations were assessed by separation on PAGE of digests (by Snap1 and Bcl-I, for Cys282Tyr and His63Asp, respectively) of
Cys282Tyr: I&l; His63Asp: HHA;wt: wild type; median, (range) HLA-H mutations were more frequent in CH-C with Tf%>45 (17/34=50%) (vs. 13/64=13% in Tf??<45; p1.9 was observed only in 1 of the 4 Cys282Tyr (others: 0.26, 0.61, 0.8) and in 1 of the 3 His63Asp homozygotes. Conclusions: Genetic factors contribute to hepatic iron accumulation in CH-C. Furthermore, Cys282Tyr or His63Asp homozygosity is not necessarily associated with a high hepatic iron content.
SWEDISH PATIENTS WITH HEMOCHROMATOSIS (HH) HAVE LOW LEVELS OF IRON AND A LOW FREQUENCY OF CIRRHOSIS
K. Hagen’, R. Befrits’, E. M. P. Cardoso’, P. Sti*, R. Hultcrantz’. ‘Department of Gastroenterology and Hepatology, Karolinska Hospital, Stockholm, *Department of Gastroenterology and Hepatology, Huddinge Univ. Hospital, Huddinge, Sweden. In recent reports, the frequency of liver cirrhosis in patients with HH is 60-70%. Liver cirrhosis is caused bv hick liver iron stores. due to late diagnosis or a more intense exbresiion of the disease. Cirrhosis may also he caused by coexisting factors, such as hepatitis B or C virus infection, or alcohol overconsumption. We studies these factors in a Swedish HH oatient erouo. The group consisted of 76 patients (56 men &d 17 women) with HH, diagnosed between 1980 and 1996 in Stockholm. All patients were homozygous for the C282Y mutation. 9 patients (11.8%) had cirrhosis, 23 (30.3%) had fibrosis and 44 (57.9%) had no fibrosis. The mean iron removed was 6.71 f 4.1 g (0.9-23.4 g), 7.2 f 4.3 g (1.4-23.4 g) for men and 5.0 f 2.9 g (0.9-14.0 g) for women. 30 patients (39.5%) had less than 5 g of removable iron. AU patients ;vith cirrhosis had iron stores exceeding 7 g. Of the patients without fibrosis, only 7 patients (15.9%) had iron stores exceeding 7 g. Iron stores were not age correlated. The mean aee at diagnosis was 47.4 f 13.5 yea& (20-73 years), 46.9 f 13.3 years (20-73 years) for men and 50.2 f 13.9 years (25-72 years) for women. 2 patients (males) were infected with hepatitis B virus, no patient with hepatitis C virus. 5 of the patients (3 men and 2 women) consumed more than 80 g alcohol/day. Conclusion: Swedish HH patients have a low frequency of cmhosls. rfijljs is not due to younger age at diagnosis, but is due to lower iron stores and a lower frequency of coexisting liver damaging factors.
REDOX GENE THERAPY OF LIVER ISCHEMIA/ REPERFUSION INJURY REDUCES AP-1 AND NF-KB ACTIVATION
Ralf M. Zwacka2, Yulong Zhanalt, Jeff Halldorsonl2, Lorita Dudus 12,and John F. Enaelhardt 1 1 University
of Iowa, Department
of Anatomy
and Cell
2 University of Edinburgh, Dept. Biology, Iowa City, Iowa. of Surgery, MRC Human Genetics Unit, Western General Hospital, Edinburgh. Liver transplantation is the only therapeutic strategy for numerous inherited and acquired diseases. The formation of reactive oxygen species (ROS) following ischemia/reperfusion (l/R) have been hypothesized to play a major role in transplant associated hepatocellular injury and ultimately graft rejection. Here, we describe the application of mitochondrial manganese superoxide dismutase (MnSOD) gene transfer to the liver as a therapeutic strategy for I/R injury in the liver of mice. Adenoviral-mediated overexpression of MnSOD in the liver of nulnu mice prior to lobar l/R achieved a 50% inhibition of acute liver damage. Associated activation of both NF-KB and AP-1 was also attenuated in animals expressing recombinant MnSOD but not in those expressing an irrelevant transgene pgalactosidase. These findings suggest that changes in the cellular redox state regulate the activity of transcription factors that are involved in the acute cellular response to I/R damage. These studies provide the foundation for redox mediated gene therapies for I/R injury which has been associated with acute rejection in autologous liver transplantation.
Topi E, k I. Tdija A,.I&ik7,,cekadaS. Antoljak N, .$tefanoviCM Clinical timte of Chem&ry and Department of pediatrics, !&he milosrdnice Univaity Hospital and S&ool of Medicine University of a=b, Kantrids Chikhen’s Hopital Rijcka University Hospital &&a, Croatia Deficiency alleles of al-anti~psin (alAT) are associated with low semm levels of alAT and can be either linkcd to liver disease and/or to early w emphysema. The most common deficiency alleles are PiS and Z. PiZ genotype is charade&cd hy G+A substition in exon 5 and change of Glu to Lys at aminoacid position 342. PiS genotype is ckuacterkd by substitotion of base A-T in exon 3 and aminmcid change of Glu to Val at p&ion 264. The aim of this study was to investigate the conco&nce between phm and genotype of al antimine polytnorpbic alleles Pi S snd Z using mutant primers in FCR-RFLp analysis. In four families (mother, father and child) phenotypes wa dctemkcd by iswlearic focusing (Pbannacia, PAG-plate pH 4D5.0) and gewypes by KR-RFLP, using mutant pimers (MWG-Biotech) for FCR, Taq I @&a) for amplicon dig&on, and 3% Meta Phor agamsc @MC Bio plodud) for electmphotetic sepamtion of restiction 6agtnerl.s. The serum alAT concentration was measured by RLD methoci.FamilyAwas~MZandMZphenotypeandgenotypeforfather motbcr and chid. Each member of this family had alAT cotwntmtion in normal ~ge,FamilyBwas~~22phenotype~genotypefor~,mmotherand child @vely. only the child in this family has madwlly decreased alAT concentration. In family C. disnepancy between @cnw-gcndypc was found: a boyptwentedZZphnwtypebuthJZgenotype.Hismc#lierwasMMandfather MZ gehotypc and phenotype. Tk cowenhation of alAT in each member of family C was in normal range. Results of family C reassessment indkatai that the discrepancy was not due to sample miskd@ or technical difficulties but a hue discordance,FamilyDwas~MZand22phenotypeandgenotypeforfather, mother and child rcspe&ely. The alAT CQncentration in this family was markcdly derreased only in child. The diw between genotype aal phenotype should he thomughly studied to identify the possible caw oP changes wzcuning along the gene to pmtein cycle.
HLA-H MUTATIONS IN PATDENTSWITH CHRONIC HEPATITIS C.
P. Ferenci. Ch. Datz. L. Kazemi-Shirazi. W. Ha&l. Th. MaierDobersberger. C.Polli. K. Kaserer. Ch. Osterreicher, P. Steindl, F. Wrba, E. Penner. Vienna, Linz, and Salzburg, Austria To determine a role of genetic hemochromatosis for iron accumulation in chronic hepatitis C (CH-C) the mutations of the HLA-H gene (Cys282Tyr, His63Asp) were evaluated in 98 (70 male/28 female) patients with biopsy-confirmed CH-C. Transferrin saturation (Tf?/o) was < 45% in 64, and >45 (range: 46-89) in 34. Hepatic iron content was measured by atom absorption spectroscopy and the hepatic iron index (I-III) was calculated. Mutations were assessed by separation on PAGE of digests (by Snap1 and Bcl-I, for Cys282Tyr and His63Asp, respectively) of
Cys282Tyr: I&l; His63Asp: HHA;wt: wild type; median, (range) HLA-H mutations were more frequent in CH-C with Tf%>45 (17/34=50%) (vs. 13/64=13% in Tf??<45; p