Fas-mediated cholangiopathy in the murine model of graft versus host disease.
Bile-duct injury observed in hepatic graft versus host disease (GVHD) is regarded as an immune-mediated injury, although its precise mechanism is unclear. However, recent studies have suggested the involvement of Fas-mediated cell death in this immune-mediated cholangiopathy. In this study, we first showed the constitutive expression of Fas receptor by cholangiocytes in situ from normal BALB/c mice, which was upregulated in GVHD mice. Also, we confirmed the Fas protein expression in the isolated cholangiocytes from normal BALB/c mice by immunocytochemistry and immunoblotting. Furthermore, the addition of agonistic Fas antibody-(Jo2)-induced cholangiocyte apoptosis confirmed by DNA-ladder formation and annexin V staining. Cholangiocytes from Fas-deficient mice (MRL lpr/lpr) did not show Jo2-induced apoptosis. Interferon-gamma augmented Fas expression and Fas-mediated cell death, respectively. Following these observations, experimental GVHD was induced by transfer of splenocytes from B10.D2 mice to irradiated (800 rad) BALB/c mice. Liver-infiltrating lymphocytes from the recipient showed dose-dependent cytotoxicity against (51)Cr-labeled cholangiocytes isolated from BALB/c mice. Moreover, the addition of blocking Fas-Fc fusion protein reduced this cytotoxicity to 44.7%. Finally, administration of this Fas-Fc protein to the BALB/c mice, which had been adoptively transferred with splenocytes of B10.D2 mice, prevented the development of hepatic GVHD in vivo. These results showed the involvement of Fas-mediated cell death in cholangiopathy observed in GVHD, and a soluble Fas-Fc protein may have a therapeutic potential for hepatic GVHD.
Ueno
Y
Y
Third Department of Internal Medicine, Tohoku University School of Medicine, Sendai, Japan. yueno@int3.med.tohoku.ac.jp
Ishii
M
M
Yahagi
K
K
Mano
Y
Y
Kisara
N
N
Nakamura
N
N
Shimosegawa
T
T
Toyota
T
T
Nagata
S
S
D000818:Animals; D001649:Bile Duct Diseases / Q000276:immunology*; D001652:Bile Ducts / Q000737:chemistry / Q000276:immunology; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic / Q000187:drug effects; D004195:Disease Models, Animal; D006086:Graft vs Host Disease; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000494:pharmacology; D008099:Liver / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D013154:Spleen / Q000637:transplantation; D019014:fas Receptor / Q000032:analysis / Q000235:genetics / Q000502:physiology*
D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; D019014:fas Receptor