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Immunohistochemical detection of polyductin and co-localization with liver progenitor cell markers during normal and abnormal development of the intrahepatic biliary system and in adult hepatobiliary carcinomas.

Dorn L, Menezes LF, Mikuz G, Otto HF, Onuchic LF, Sergi C - J. Cell. Mol. Med. (2008)

Bottom Line: No specific staining was found at the stage of remodelled bile ducts.Polyductin was also co-localized in some DP cells together with oval stem cell markers.These results represent the first systematic study of polyductin expression in human pathologies associated with abnormal development of intrahepatic biliary tree, and support the following conclusions: (i) polyductin expression mirrors developmental properties of the primitive intrahepatic biliary system; (ii) polyductin is re-expressed in pathological conditions associated with DPM and (iii) polyductin might be a potential marker to distinguish CCC from HCC.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology, University of Innsbruck, Innsbruck, Austria.

ABSTRACT
The longest open reading frame of PKHD1 (polycystic kidney and hepatic disease 1), the autosomal recessive polycystic kidney disease (ARPKD) gene, encodes a single-pass, integral membrane protein named polyductin or fibrocystin. A fusion protein comprising its intracellular C-terminus, FP2, was previously used to raise a polyclonal antiserum shown to detect polyductin in several human tissues, including liver. In the current study, we aimed to investigate by immunohistochemistry the detailed polyductin localization pattern in normal (ductal plate [DP], remodelling ductal plate [RDP], remodelled bile ducts) and abnormal development of the primitive intrahepatic biliary system, known as ductal plate malformation (DPM). This work also included the characterization of polyductin expression profile in various histological forms of neonatal and infantile cholestasis, and in cholangiocellular carcinoma (CCC) and hepatocellular carcinoma (HCC). We detected polyductin expression in the intrahepatic biliary system during the DP and the RDP stages as well as in DPM. No specific staining was found at the stage of remodelled bile ducts. Polyductin was also detected in liver biopsies with neonatal cholestasis, including mainly biliary atresia and neonatal hepatitis with ductular reaction as well as congenital hepatic fibrosis. In addition, polyductin was present in CCC, whereas it was absent in HCC. Polyductin was also co-localized in some DP cells together with oval stem cell markers. These results represent the first systematic study of polyductin expression in human pathologies associated with abnormal development of intrahepatic biliary tree, and support the following conclusions: (i) polyductin expression mirrors developmental properties of the primitive intrahepatic biliary system; (ii) polyductin is re-expressed in pathological conditions associated with DPM and (iii) polyductin might be a potential marker to distinguish CCC from HCC.

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Polyductin expression in liver disease of newborns and infants. In (A), there is liver tissue with a baseline staining (anti-FP2, ×100) with no staining of the interlobular bile duct (arrow) (inset, anti-FP2, ×320); (B) and (C) show two examples of biliary atresia with ductular proliferation without ductal plate malformation at early (B) (the arrow points to a bile duct) and advanced (C) stage showing the ductular proliferation (arrowheads) of biliary atresia with a moderate FP2 staining (anti-FP2, ×400); (D) and (E) show two examples of biliary atresia with ductal plate malformation showing biliary structures (arrowheads) at early (D) and advanced (E) stage showing moderate FP2 staining (anti-FP2, d:×400, e:×200); (F) and (G) show liver tissue affected with neonatal hepatitis with mostly an intense staining of both pseudorosettes and neoductuli (arrows with continuous line), whereas giant cells show mostly very faint staining (arrow with dotted line) (anti-FP2, ×400). In (H) is shown liver tissue from an infant with congenital liver fibrosis (note the lack of ductular proliferation at the edges of the portal tracts) showing moderate diffuse FP2 staining of the biliary structures, whereas the hepatocytes show a mild diffuse staining. The arrows point to a selected region shown in inset. Note the enlarged biliary structure with moderate FP2 staining (anti-FP2, ×200 and ×320 in the inset).
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fig02: Polyductin expression in liver disease of newborns and infants. In (A), there is liver tissue with a baseline staining (anti-FP2, ×100) with no staining of the interlobular bile duct (arrow) (inset, anti-FP2, ×320); (B) and (C) show two examples of biliary atresia with ductular proliferation without ductal plate malformation at early (B) (the arrow points to a bile duct) and advanced (C) stage showing the ductular proliferation (arrowheads) of biliary atresia with a moderate FP2 staining (anti-FP2, ×400); (D) and (E) show two examples of biliary atresia with ductal plate malformation showing biliary structures (arrowheads) at early (D) and advanced (E) stage showing moderate FP2 staining (anti-FP2, d:×400, e:×200); (F) and (G) show liver tissue affected with neonatal hepatitis with mostly an intense staining of both pseudorosettes and neoductuli (arrows with continuous line), whereas giant cells show mostly very faint staining (arrow with dotted line) (anti-FP2, ×400). In (H) is shown liver tissue from an infant with congenital liver fibrosis (note the lack of ductular proliferation at the edges of the portal tracts) showing moderate diffuse FP2 staining of the biliary structures, whereas the hepatocytes show a mild diffuse staining. The arrows point to a selected region shown in inset. Note the enlarged biliary structure with moderate FP2 staining (anti-FP2, ×200 and ×320 in the inset).

Mentions: None or very faint FP2 staining was noted in the hepatocytes or biliary structures of unaffected liver tissue and FP2 immunostained slides did not show any signal with regard to the canals of Hering. Figure 2A shows liver tissue with a baseline staining. In five out of ten cases with BA without DPM at early and at advanced stage with ductular proliferation (Fig. 2B and C), there was a faint staining of the portal ductal structures (early stage, Fig. 2B) and moderate FP2 staining of the biliary proliferations of the portal tracts (advanced stage, Fig. 2C). In three additional cases of BA with DPM at early (Fig. 2D) and at advanced stage (Fig. 2E) with ductular proliferation, FP2 staining of the biliary structures was strong. In two cases of BA without DPM, no staining was detected. NH showed a moderate to strong staining of the liver tissue in six out of nine cases (Fig. 2F and G), whereas a mild staining was found in three cases. In particular, pseudorosettes and neoductuli showed mostly an intense FP2 staining, whereas giant cells showed mostly very faint staining. Liver tissue from two patients with CHF (Fig. 2H) showed a moderate diffuse staining of the ductular structures, whilst the hepatocytes showed a mild diffuse staining. In liver biopsies with paucity of the intrahepatic bile ducts (PIBD) (Fig. 3A), intense, mild and no FP2 staining were found in two, nine and five cases, respectively. Finally, a mild staining was found in single cases of sepsis, choledocal cyst-associated liver disease (CC-LD), annular pancreas obstruction-associated liver disease (APO-LD), cardiovascular disease with hepatic involvement (CVD-HI), FFAOD (foetal fatty acid oxidation disorder associated with maternal HELLP syndrome), drug-induced liver disease (DILD), non-biliary liver disease (NBLD NOS), and liver disease associated with necrotizing enterocolitis (NEC-LD) as well as in two cases of neonatal cholestasis not otherwise specified (NC NOS). An additional case of an infant with alpha-1-antitrypsin deficiency (Fig. 3B) was added to the study and showed more intense FP2 staining at the periportal area. The liver tissue of progressive familiar intrahepatic cholestasis (Fig. 3C) showed an intense staining of the biliary structures and periportal hepatocytes. In Caroli’s disease (Fig. 3D), there were intraluminal bulbar protrusions of the ductal wall with moderate to intense staining of the biliary structures. The other cases of Table 1 did not show any staining. No age-dependent differences were found within each group.


Immunohistochemical detection of polyductin and co-localization with liver progenitor cell markers during normal and abnormal development of the intrahepatic biliary system and in adult hepatobiliary carcinomas.

Dorn L, Menezes LF, Mikuz G, Otto HF, Onuchic LF, Sergi C - J. Cell. Mol. Med. (2008)

Polyductin expression in liver disease of newborns and infants. In (A), there is liver tissue with a baseline staining (anti-FP2, ×100) with no staining of the interlobular bile duct (arrow) (inset, anti-FP2, ×320); (B) and (C) show two examples of biliary atresia with ductular proliferation without ductal plate malformation at early (B) (the arrow points to a bile duct) and advanced (C) stage showing the ductular proliferation (arrowheads) of biliary atresia with a moderate FP2 staining (anti-FP2, ×400); (D) and (E) show two examples of biliary atresia with ductal plate malformation showing biliary structures (arrowheads) at early (D) and advanced (E) stage showing moderate FP2 staining (anti-FP2, d:×400, e:×200); (F) and (G) show liver tissue affected with neonatal hepatitis with mostly an intense staining of both pseudorosettes and neoductuli (arrows with continuous line), whereas giant cells show mostly very faint staining (arrow with dotted line) (anti-FP2, ×400). In (H) is shown liver tissue from an infant with congenital liver fibrosis (note the lack of ductular proliferation at the edges of the portal tracts) showing moderate diffuse FP2 staining of the biliary structures, whereas the hepatocytes show a mild diffuse staining. The arrows point to a selected region shown in inset. Note the enlarged biliary structure with moderate FP2 staining (anti-FP2, ×200 and ×320 in the inset).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4496142&req=5

fig02: Polyductin expression in liver disease of newborns and infants. In (A), there is liver tissue with a baseline staining (anti-FP2, ×100) with no staining of the interlobular bile duct (arrow) (inset, anti-FP2, ×320); (B) and (C) show two examples of biliary atresia with ductular proliferation without ductal plate malformation at early (B) (the arrow points to a bile duct) and advanced (C) stage showing the ductular proliferation (arrowheads) of biliary atresia with a moderate FP2 staining (anti-FP2, ×400); (D) and (E) show two examples of biliary atresia with ductal plate malformation showing biliary structures (arrowheads) at early (D) and advanced (E) stage showing moderate FP2 staining (anti-FP2, d:×400, e:×200); (F) and (G) show liver tissue affected with neonatal hepatitis with mostly an intense staining of both pseudorosettes and neoductuli (arrows with continuous line), whereas giant cells show mostly very faint staining (arrow with dotted line) (anti-FP2, ×400). In (H) is shown liver tissue from an infant with congenital liver fibrosis (note the lack of ductular proliferation at the edges of the portal tracts) showing moderate diffuse FP2 staining of the biliary structures, whereas the hepatocytes show a mild diffuse staining. The arrows point to a selected region shown in inset. Note the enlarged biliary structure with moderate FP2 staining (anti-FP2, ×200 and ×320 in the inset).
Mentions: None or very faint FP2 staining was noted in the hepatocytes or biliary structures of unaffected liver tissue and FP2 immunostained slides did not show any signal with regard to the canals of Hering. Figure 2A shows liver tissue with a baseline staining. In five out of ten cases with BA without DPM at early and at advanced stage with ductular proliferation (Fig. 2B and C), there was a faint staining of the portal ductal structures (early stage, Fig. 2B) and moderate FP2 staining of the biliary proliferations of the portal tracts (advanced stage, Fig. 2C). In three additional cases of BA with DPM at early (Fig. 2D) and at advanced stage (Fig. 2E) with ductular proliferation, FP2 staining of the biliary structures was strong. In two cases of BA without DPM, no staining was detected. NH showed a moderate to strong staining of the liver tissue in six out of nine cases (Fig. 2F and G), whereas a mild staining was found in three cases. In particular, pseudorosettes and neoductuli showed mostly an intense FP2 staining, whereas giant cells showed mostly very faint staining. Liver tissue from two patients with CHF (Fig. 2H) showed a moderate diffuse staining of the ductular structures, whilst the hepatocytes showed a mild diffuse staining. In liver biopsies with paucity of the intrahepatic bile ducts (PIBD) (Fig. 3A), intense, mild and no FP2 staining were found in two, nine and five cases, respectively. Finally, a mild staining was found in single cases of sepsis, choledocal cyst-associated liver disease (CC-LD), annular pancreas obstruction-associated liver disease (APO-LD), cardiovascular disease with hepatic involvement (CVD-HI), FFAOD (foetal fatty acid oxidation disorder associated with maternal HELLP syndrome), drug-induced liver disease (DILD), non-biliary liver disease (NBLD NOS), and liver disease associated with necrotizing enterocolitis (NEC-LD) as well as in two cases of neonatal cholestasis not otherwise specified (NC NOS). An additional case of an infant with alpha-1-antitrypsin deficiency (Fig. 3B) was added to the study and showed more intense FP2 staining at the periportal area. The liver tissue of progressive familiar intrahepatic cholestasis (Fig. 3C) showed an intense staining of the biliary structures and periportal hepatocytes. In Caroli’s disease (Fig. 3D), there were intraluminal bulbar protrusions of the ductal wall with moderate to intense staining of the biliary structures. The other cases of Table 1 did not show any staining. No age-dependent differences were found within each group.

Bottom Line: No specific staining was found at the stage of remodelled bile ducts.Polyductin was also co-localized in some DP cells together with oval stem cell markers.These results represent the first systematic study of polyductin expression in human pathologies associated with abnormal development of intrahepatic biliary tree, and support the following conclusions: (i) polyductin expression mirrors developmental properties of the primitive intrahepatic biliary system; (ii) polyductin is re-expressed in pathological conditions associated with DPM and (iii) polyductin might be a potential marker to distinguish CCC from HCC.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology, University of Innsbruck, Innsbruck, Austria.

ABSTRACT
The longest open reading frame of PKHD1 (polycystic kidney and hepatic disease 1), the autosomal recessive polycystic kidney disease (ARPKD) gene, encodes a single-pass, integral membrane protein named polyductin or fibrocystin. A fusion protein comprising its intracellular C-terminus, FP2, was previously used to raise a polyclonal antiserum shown to detect polyductin in several human tissues, including liver. In the current study, we aimed to investigate by immunohistochemistry the detailed polyductin localization pattern in normal (ductal plate [DP], remodelling ductal plate [RDP], remodelled bile ducts) and abnormal development of the primitive intrahepatic biliary system, known as ductal plate malformation (DPM). This work also included the characterization of polyductin expression profile in various histological forms of neonatal and infantile cholestasis, and in cholangiocellular carcinoma (CCC) and hepatocellular carcinoma (HCC). We detected polyductin expression in the intrahepatic biliary system during the DP and the RDP stages as well as in DPM. No specific staining was found at the stage of remodelled bile ducts. Polyductin was also detected in liver biopsies with neonatal cholestasis, including mainly biliary atresia and neonatal hepatitis with ductular reaction as well as congenital hepatic fibrosis. In addition, polyductin was present in CCC, whereas it was absent in HCC. Polyductin was also co-localized in some DP cells together with oval stem cell markers. These results represent the first systematic study of polyductin expression in human pathologies associated with abnormal development of intrahepatic biliary tree, and support the following conclusions: (i) polyductin expression mirrors developmental properties of the primitive intrahepatic biliary system; (ii) polyductin is re-expressed in pathological conditions associated with DPM and (iii) polyductin might be a potential marker to distinguish CCC from HCC.

Show MeSH
Related in: MedlinePlus