Immunohistochemical localization of transforming growth factor β-1 and its relationship with collagen expression in advanced liver fibrosis due to biliary atresia

Abstract

Purpose:

Biliary atresia (BA) is the most common indication of liver transplantation in children. Pathogenesis of hepatic fibrosis, which is a prominent feature of BA, remains obscure. The purpose of this work was to determine the cellular sources of transforming growth factor beta-1 (TGFβ1) and establish the relationship between TGFβ1-producing cells and extracellular matrix producing myofibroblasts (MFBs) in advanced BA.

Methods:

Trichrome staining and immunohistochemistry were carried out to determine the expression pattern of collagen and TGFβ1 protein in explant liver specimens from patients with BA. The intensities of portal and lobular TGFβ1 expressions were compared. Immunofluorescence technique was carried out to determine the relationship between α-smooth muscle actin (α-SMA)-positive-MFB and TGFβ1-positve cells.

Results:

Lobular TGFβ1 protein expression was significantly higher than portal (89 ± 6 versus 10 ± 1 arbitrary units, P ≤ 0.05), whereas no difference was noted in livers used as control (10 ± 1.6 versus 19 ± 5 arbitrary units, P = 0.11). TGFβ1 expression was more in the center of nodules versus MFB in surrounding fibrous septa. Contrary to TGFβ1 expression, α1-SMA was mostly expressed in the portal structures and the adjacent fibrous septa enacting lobulation of the parenchyma. The results obtained by coimmunofluorescence staining showed no colocalization of α-SMA and TGFβ1.

Conclusions:

TGFβ1 protein expression is mostly localized to hepatocytes in advanced BA. These findings suggest a paracrine mechanisms of TGFβ1-driven fibrogenesis in advanced BA.

Keywords:

• biliary atresia
• liver fibrosis
• transforming growth factor β-1
• liver transplantation

Acknowledgements

This work was supported by funds from the Children’s Miracle Network. Publication of this article was funded in part by the University of Florida Open-Access Publishing Fund. The authors thank Dr Bryon E Petersen for his expert input in the analysis of immunofluorescence and for reviewing the manuscript. We also thank Marda Jorgensen for technical advice on immunohistochemistry.

Disclosure

The authors report no conflicts of interest in this work.