Patterns of intermediate filaments, VLA integrins and HLA antigens in a new human biliary epithelial cell line sensitive to interferon-gamma.
OBJECTIVE
Intra-hepatic bile ducts are the primary site of damage in several immunologically mediated liver diseases. However, immunological processes underlying biliary epithelial cell recognition by T lymphocytes are poorly understood. Therefore, a convenient in vitro model that could mimic these immunologic disorders would be of great interest.


METHODS
A human cell line (HuGB) was established from a metastasis of gallbladder adenocarcinoma in the liver. Intermediate filament expression was analysed by immunostaining, and gamma-glutamyl transpeptidase and albumin secretion were measured. VLA integrin expression pattern, expression of HLA class I and II antigens and ICAM-1 protein were analysed by flow cytometry and their modulation by interferon-gamma was quantitated using a QIFIKIT commercial kit.


RESULTS
Histological analysis showed high similarity between the initial gallbladder adenocarcinoma and the established cell line. Cytokeratins 8 and 19 and vimentin showed strong positive staining in the established cell line. Gamma-glutamyl transpeptidase was secreted by these cells while albumin expression was negative. HuGB cells also expressed VLA-alpha2, VLA-alpha3, VLA-alpha6, VLA-beta1, but not VLA-alpha1, VLA-alpha4 and NCAM, a pattern of adhesion molecule expression compatible with the biliary epithelium. Also, similar to the biliary epithelium found in normal liver, HuGB cells expressed abundant HLA class I but few HLA class II antigens. We found that the expression of HLA antigens and ICAM-1 protein were increased during interferon-gamma treatment of HuGB cell line.


CONCLUSIONS
Both phenotypic and morphological characteristics of HuGB cells suggested their biliary origin. Sensitivity of HuGB cells to interferon-gamma suggests that this new cell line could represent a suitable model to investigate the up-regulation of membrane antigens occurring in immune diseases involving biliary epithelial cells.
Rumin
S
S
INSERM U 49, Unité de Recherches Hépatologiques, CHRU Pontchaillou, Rennes, France. rumin@lyon151.inserm.fr
Loréal
O
O
Drénou
B
B
Turlin
B
B
Rissel
M
M
Campion
J P
JP
Gripon
P
P
Strain
A J
AJ
Clément
B
B
Guguen-Guillouzo
C
C
D000230:Adenocarcinoma; D000368:Aged; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D002466:Cell Nucleolus / Q000648:ultrastructure; D002467:Cell Nucleus / Q000648:ultrastructure; D004848:Epithelium / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D005704:Gallbladder / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D005706:Gallbladder Neoplasms; D006681:HLA-D Antigens / Q000032:analysis / Q000096:biosynthesis; D015395:Histocompatibility Antigens Class I / Q000032:analysis / Q000096:biosynthesis; D006801:Humans; D007150:Immunohistochemistry; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D007371:Interferon-gamma / Q000494:pharmacology*; D007381:Intermediate Filament Proteins / Q000032:analysis / Q000096:biosynthesis*; D008099:Liver / Q000276:immunology / Q000502:physiology; D008297:Male; D008871:Microvilli / Q000648:ultrastructure; D008928:Mitochondria / Q000648:ultrastructure; D016029:Receptors, Very Late Antigen / Q000032:analysis / Q000096:biosynthesis*; D014617:Vacuoles / Q000648:ultrastructure
D006681:HLA-D Antigens; D015395:Histocompatibility Antigens Class I; D007381:Intermediate Filament Proteins; D016029:Receptors, Very Late Antigen; D018799:Intercellular Adhesion Molecule-1; D007371:Interferon-gamma