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Perisinusoidal fibrogenesis and ductular metaplasia of hepatocytes in focal nodular hyperplasia of the liver
59
EXPRESSION OF PRE-S GI~E-~I~CODED PROTEINS IN LIVER AND SERUM DURING CHRONIC HEPATITIS B VIRUS INFECTION. S.J.Hadziyarsnis, G.Raimondo, Ch.Papaioannou, Ch.Anastassakos, D.Wong, J.Sinsky and D.A. S~afritz, Hippokrat"-~----'-~io~ --~" Gener-----~Hosp------ital,Athens, Greece ,------~rt ----=~---~-in Elnste" College of Medicine, N.Y. and Cetus Corp., Emeryville, CA., USA.
Pre-S gene-encoded proteins of the hepatitis B virus (HBV) were studied in the liver by immunofluorescence and in serum by radioimmunoassay in 30 patients with chronic HBV infection. The results were compared with molecular hybridization analysis of HBV-DNA in liver and serum, with serum hepatitis B e antigen/antibody (HBeAg/anti-HBe) status and with underlying liver histology. Pre-S peptides were detected in the s ~ of 11 patients, I0 of whom were positive for s ~ HBV-DNA and/or liver hepatitis B core antigen. Only 4 of these patients were HBeAg positive. The prevalence of sert~ pre-S among HBV replicating carriers was 59% (10/17) compared to only 8% (I/13) among those with non-replicating virus (p<0.001) . All patients with circulating pre-S peptides had active liver disease. Anti-pre-S was detected in the serum of only 4 patients, 3 with integrated HBV-E~A. In contrast to serum findings, pre-S peptides were detected in the liver of all patients with histochemically demonstrable hepatitis B surface antigen (HBsAg) regardless of HBV replicative status. HBsAg carriers with integrated HBV-ENA had abundant cytoplasmic pre-S localized in numerous ground-glass hepatocytes. It is concluded that I) pre-S peptides are expressed in the liver whenever HBsAg synthesis occurs, 2) they are secreted in the serum in association with high HBV replication and release of HBV particles and 3) in the absence of episcmal HBV replication, pre-S peptides seem to be retained within hepatocytic membranes. Consistent with studies in cell culture systems, these results suggest a pathway through a membranous system in assembly or secretion of HBV particles and that pre-S epltopes play an important role in this process.
60
PERISINUSOIDAL FIBROGENESIS AND DUCTULAR METAPLASIA OF HEPATOCYTES IN FOCAL NODULAR HYPERPLASIA OF THE LIVER. E.G. Hahn, 3. Kirchner'. D. Schuooan', Dept. of Medicine, Thomas Jefferson University, Philadelphia, USA and "Dept. of Medicine, Steglltz Medical School, Free University, West Berlin, FRG
We have investigated the composition of perisinusoidal fibrosis (PSF) in its relation to ductular metaplasia of hepatocytes (DMH) in focal nodular hyperplasia of the l i v e r (FNH). 4~m frozen, unfixed sections of FNH tissue (n=4) were stained by indirect immunofluorescence. Antibodies against human f i b r i l l a r collagens I , I I I , V and VI, basement membrane collagen IV, basement membrane glycoprotein l amlnln and fibronectin were produced in rabblts and goats, purified by immunoabsorption and used in a concentration of IO-5Ogg IgG/ml. Monoclonal antibodies to cytokeratin from Dakopatts, Hamburg, were used in double-stalnlng experiments to visualize OMH. All connective tissue proteins are shown in the perisinusoidal spaces of hyperplastic nodules and in fibrotic areas. A most conspicuous increase of the basement membrane proteins collagen IV and lamlnin and collagen V is observed in islands of bile ductule "proliferation" with a gradient from non-fibrotic perisinusoldal spaces outside to the inner accumulation of bile ductules. A similar gradient exists for cytokeratln-positlve cells, which appear inside the zone of PSF where cholestasls and DMH are known to occur (Butron Vila et al., Liver 4:387, 1984). Periductular fibrosis and fibrous septa display a similar composition. Outside vascular structures, a vast preponderance of fibrous collagens I , I I I and Vl is noted with collagens IV, V and l amlnin interspersed. This pattern resembles a matrix produced by smooth muscle cells and may be related to myofibroblasts (Callea et a l . , Virch Arch A 396:155, 1982). Since cytokeratin-positive cells constantly appear in close contact to PSF, we conclude that the induction of a new juxtahepatocellular matrix (probably related to myofibroblasts) may permit the i n i t i a t i o n of DMH. FNH may serve as a model to elucidate the sequential interplay of genes related to connective tissue production and to changes of epithelial phenotype.
S32
EXPRESSION OF PRE-S GI~E-~I~CODED PROTEINS IN LIVER AND SERUM DURING CHRONIC HEPATITIS B VIRUS INFECTION. S.J.Hadziyarsnis, G.Raimondo, Ch.Papaioannou, Ch.Anastassakos, D.Wong, J.Sinsky and D.A. S~afritz, Hippokrat"-~----'-~io~ --~" Gener-----~Hosp------ital,Athens, Greece ,------~rt ----=~---~-in Elnste" College of Medicine, N.Y. and Cetus Corp., Emeryville, CA., USA.
Pre-S gene-encoded proteins of the hepatitis B virus (HBV) were studied in the liver by immunofluorescence and in serum by radioimmunoassay in 30 patients with chronic HBV infection. The results were compared with molecular hybridization analysis of HBV-DNA in liver and serum, with serum hepatitis B e antigen/antibody (HBeAg/anti-HBe) status and with underlying liver histology. Pre-S peptides were detected in the s ~ of 11 patients, I0 of whom were positive for s ~ HBV-DNA and/or liver hepatitis B core antigen. Only 4 of these patients were HBeAg positive. The prevalence of sert~ pre-S among HBV replicating carriers was 59% (10/17) compared to only 8% (I/13) among those with non-replicating virus (p<0.001) . All patients with circulating pre-S peptides had active liver disease. Anti-pre-S was detected in the serum of only 4 patients, 3 with integrated HBV-E~A. In contrast to serum findings, pre-S peptides were detected in the liver of all patients with histochemically demonstrable hepatitis B surface antigen (HBsAg) regardless of HBV replicative status. HBsAg carriers with integrated HBV-ENA had abundant cytoplasmic pre-S localized in numerous ground-glass hepatocytes. It is concluded that I) pre-S peptides are expressed in the liver whenever HBsAg synthesis occurs, 2) they are secreted in the serum in association with high HBV replication and release of HBV particles and 3) in the absence of episcmal HBV replication, pre-S peptides seem to be retained within hepatocytic membranes. Consistent with studies in cell culture systems, these results suggest a pathway through a membranous system in assembly or secretion of HBV particles and that pre-S epltopes play an important role in this process.
60
PERISINUSOIDAL FIBROGENESIS AND DUCTULAR METAPLASIA OF HEPATOCYTES IN FOCAL NODULAR HYPERPLASIA OF THE LIVER. E.G. Hahn, 3. Kirchner'. D. Schuooan', Dept. of Medicine, Thomas Jefferson University, Philadelphia, USA and "Dept. of Medicine, Steglltz Medical School, Free University, West Berlin, FRG
We have investigated the composition of perisinusoidal fibrosis (PSF) in its relation to ductular metaplasia of hepatocytes (DMH) in focal nodular hyperplasia of the l i v e r (FNH). 4~m frozen, unfixed sections of FNH tissue (n=4) were stained by indirect immunofluorescence. Antibodies against human f i b r i l l a r collagens I , I I I , V and VI, basement membrane collagen IV, basement membrane glycoprotein l amlnln and fibronectin were produced in rabblts and goats, purified by immunoabsorption and used in a concentration of IO-5Ogg IgG/ml. Monoclonal antibodies to cytokeratin from Dakopatts, Hamburg, were used in double-stalnlng experiments to visualize OMH. All connective tissue proteins are shown in the perisinusoidal spaces of hyperplastic nodules and in fibrotic areas. A most conspicuous increase of the basement membrane proteins collagen IV and lamlnin and collagen V is observed in islands of bile ductule "proliferation" with a gradient from non-fibrotic perisinusoldal spaces outside to the inner accumulation of bile ductules. A similar gradient exists for cytokeratln-positlve cells, which appear inside the zone of PSF where cholestasls and DMH are known to occur (Butron Vila et al., Liver 4:387, 1984). Periductular fibrosis and fibrous septa display a similar composition. Outside vascular structures, a vast preponderance of fibrous collagens I , I I I and Vl is noted with collagens IV, V and l amlnin interspersed. This pattern resembles a matrix produced by smooth muscle cells and may be related to myofibroblasts (Callea et a l . , Virch Arch A 396:155, 1982). Since cytokeratin-positive cells constantly appear in close contact to PSF, we conclude that the induction of a new juxtahepatocellular matrix (probably related to myofibroblasts) may permit the i n i t i a t i o n of DMH. FNH may serve as a model to elucidate the sequential interplay of genes related to connective tissue production and to changes of epithelial phenotype.
S32