The immunology of fibrogenesis in alcoholic liver disease.
BACKGROUND
Alcoholic liver disease in humans frequently leads to cirrhosis. Experimental models of hepatic fibrogenesis are available, but extrapolation of those findings to human ethanol-induced liver injury is difficult. Hepatic ethanol-induced fibrosis in humans has often been studied in relatively small patient populations. During the past decade, several animal models and human studies have attributed fibrogenesis in the liver to the role played by hepatocytes, Kupffer cells, endothelial cells, and especially stellate cells.


OBJECTIVE
To determine the contribution of the main liver cell types to ethanol-induced fibrogenesis. For that purpose, we studied the expression of the following immunologic parameters: smooth muscle-specific alpha actin (SMSA), CD68, CD34, transforming growth factor beta1, intercellular adhesion molecule 1, and collagen types 1 and 3. The Dako LSAB+ kit (peroxidase method) was used.


METHODS
We recently studied a large cohort of patients with alcoholic liver disease in France. In this cohort, we found 87 cases in which liver biopsies revealed only pericentral injury with nonpathologic portal areas. We compared cases in which the portal areas were nonpathologic with 324 patients in whom staging ranged from F0 to F3. Patients with cirrhosis (F4) were excluded from evaluation. To stage fibrosis, we used the METAVIR system. Furthermore, we selected 40 cases in which the biopsies measured at least 25 mm in length for further histochemical evaluation. Ten additional normal cases from our archives were used as controls. We divided this patient population into the following 5 groups of 10 patients each: group 1A, F0 with steatosis; group 1B, F0 without steatosis; group 2, F0 to F1, central injury; group 3, F3, fibrosis with multiple septa; and group 4, nonpathologic livers (controls).


RESULTS
Smooth muscle-specific alpha actin was expressed by stellate cells, pericentrally, with increasing severity and intensity in the advanced stage of fibrosis of group 3, less intense expression was noted in group 2, and expression was practically absent in group 1 and in nonpathologic controls. CD68 was the best marker for Kupffer cells and was expressed diffusely within the lobules in all groups. Its expression correlated directly with the degree of disease severity, progressing from stage I through stage III, but was absent in nonpathologic livers. CD34 was consistently expressed by endothelial cells in the periportal areas in all groups. The expression of collagen type 1 was intense in the bands of fibrosis or bridging, while type 3 expression was poor. Transforming growth factor beta1 and intercellular adhesion molecule 1 were not expressed in any group.


CONCLUSIONS
In this study, stellate cell activation (SMSA) was most intense pericentrally in the early stages and diffusely with progression to fibrosis and maximum intensity in stage III. Kupffer cell activation, as determined by CD68 expression, was intense and diffuse, while endothelial cells expressed CD34 periportally in a similar manner in all stages. Fibrogenesis in human ethanol injury is due to the activity of stellate cells, Kupffer cells, and to a lesser extent, to endothelial cells.
Chedid
Antonio
A
Department of Pathology, Rosalind Franklin University/Chicago Medical School, North Chicago, Ill 60064, USA. Antonio.chedid@rosalindfranklin.edu
Arain
Selma
S
Snyder
Ann
A
Mathurin
Philippe
P
Capron
Frédérique
F
Naveau
Sylvie
S
D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D015331:Cohort Studies; D042783:Endothelial Cells / Q000737:chemistry / Q000473:pathology; D000431:Ethanol / Q000506:poisoning; D005260:Female; D022781:Hepatocytes / Q000737:chemistry / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D007728:Kupffer Cells / Q000737:chemistry / Q000502:physiology; D008103:Liver Cirrhosis / Q000209:etiology* / Q000276:immunology*; D008108:Liver Diseases, Alcoholic / Q000150:complications*; D008297:Male; D008875:Middle Aged
D015415:Biomarkers; D000431:Ethanol