title: Involvement of the TNF and FasL produced by CD11b Kupffer cells/macrophages in CCl4-induced acute hepatic injury.
abstract: We previously reported that F4/80(+) Kupffer cells are subclassified into CD68(+) Kupffer cells with phagocytic and ROS producing capacity, and CD11b(+) Kupffer cells with cytokine-producing capacity. Carbon tetrachloride (CCl4)-induced hepatic injury is a well-known chemical-induced hepatocyte injury. In the present study, we investigated the immunological role of Kupffer cells/macrophages in CCl4-induced hepatitis in mice. The immunohistochemical analysis of the liver and the flow cytometry of the liver mononuclear cells showed that clodronate liposome (c-lipo) treatment greatly decreased the spindle-shaped F4/80(+) or CD68(+) cells, while the oval-shaped F4/80+ CD11b(+) cells increased. Notably, severe hepatic injury induced by CCl4 was further aggravated by c-lipo-pretreatment. The population of CD11b(+) Kupffer cells/macrophages dramatically increased 24 hour (h) after CCl4 administration, especially in c-lipo-pretreated mice. The CD11b(+) Kupffer cells expressed intracellular TNF and surface Fas-ligand (FasL). Furthermore, anti-TNF Ab pretreatment (which decreased the FasL expression of CD11b(+) Kupffer cells), anti-FasL Ab pretreatment or gld/gld mice attenuated the liver injury induced by CCl4. CD1d-/- mouse and cell depletion experiments showed that NKT cells and NK cells were not involved in the hepatic injury. The adoptive transfer and cytotoxic assay against primary cultured hepatocytes confirmed the role of CD11b(+) Kupffer cells in CCl4-induced hepatitis. Interestingly, the serum MCP-1 level rapidly increased and peaked at six h after c-lipo pretreatment, suggesting that the MCP-1 produced by c-lipo-phagocytized CD68(+) Kupffer cells may recruit CD11b(+) macrophages from the periphery and bone marrow. The CD11b(+) Kupffer cells producing TNF and FasL thus play a pivotal role in CCl4-induced acute hepatic injury.
authors.affiliation: Department of Immunology and Microbiology, National Defense Medical College, Namiki, Tokorozawa, Saitama, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sato
authors.affiliation: Department of Immunology and Microbiology, National Defense Medical College, Namiki, Tokorozawa, Saitama, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Nakashima
authors.affiliation: Department of Immunology and Microbiology, National Defense Medical College, Namiki, Tokorozawa, Saitama, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Nakashima
authors.affiliation: Department of Immunology and Microbiology, National Defense Medical College, Namiki, Tokorozawa, Saitama, Japan.
authors.forename: Masami
authors.initials: M
authors.lastname: Ikarashi
authors.affiliation: Department of Surgery 1, National Defense Medical College, Namiki, Tokorozawa, Saitama, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Nishiyama
authors.affiliation: Department of Immunology and Microbiology, National Defense Medical College, Namiki, Tokorozawa, Saitama, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Kinoshita
authors.affiliation: Department of Immunology and Microbiology, National Defense Medical College, Namiki, Tokorozawa, Saitama, Japan.
authors.forename: Shuhji
authors.initials: S
authors.lastname: Seki
mesh_terms: D000818:Animals; D055610:Antigens, CD1d / Q000502:physiology; D002251:Carbon Tetrachloride / Q000633:toxicity*; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D053222:Fas Ligand Protein / Q000378:metabolism*; D005434:Flow Cytometry; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D007124:Immunoenzyme Techniques; D007694:Killer Cells, Natural / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D007728:Kupffer Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D017114:Liver Failure, Acute / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D055610:Antigens, CD1d; C584692:CD1d antigen, mouse; D053222:Fas Ligand Protein; D014409:Tumor Necrosis Factor-alpha; D002251:Carbon Tetrachloride