title: Gene transfer of high-mobility group box 1 box-A domain in a rat acute liver failure model.
abstract: BACKGROUND
  High-mobility group box 1 (HMGB1) has recently been identified as an important mediator of various kinds of acute and chronic inflammation. The protein encoded by the box-A domain of the HMGB1 gene is known to act as a competitive inhibitor of HMGB1. In this study, we investigated whether box-A gene transfer results in box-A protein production in rats and assessed therapeutic efficacy in vivo using an acute liver failure (ALF) model.
  
  
  METHODS
  Three types of adenovirus vectors were constructed-a wild type and two mutants-and a mutant vector was then selected based on the secretion from HeLa cells. The secreted protein was subjected to a tumor necrosis factor (TNF) production inhibition test in vitro. The vector was injected via the portal vein in healthy Wistar rats to confirm box-A protein production in the liver. The vector was then injected via the portal vein in rats with ALF.
  
  
  RESULTS
  Western blot analysis showed enhanced expression of box-A protein in HeLa cells transfected with one of the mutant vectors. The culture supernatant from HeLa cells transfected with the vector inhibited TNF-α production from macrophages. Expression of box-A protein was confirmed in the transfected liver at 72 h after transfection. Transfected rats showed decreased hepatic enzymes, plasma HMGB1, and hepatic TNF-α messenger RNA levels, and histologic findings and survival were significantly improved.
  
  
  CONCLUSIONS
  HMGB1 box-A gene transfer results in box-A protein production in the liver and appears to have a beneficial effect on ALF in rats.
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Tanaka
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan. Electronic address: masa02114@yahoo.co.jp.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Shinoda
authors.affiliation: Department of Molecular Biology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Takayanagi
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Go
authors.initials: G
authors.lastname: Oshima
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Nishiyama
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Fukuda
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yagi
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Tetsu
authors.initials: T
authors.lastname: Hayashida
authors.affiliation: Department of Pathology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Masugi
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Suda
authors.affiliation: Central Institute, Shino-Test Corporation, Sagamihara, Kanagawa, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Yamada
authors.affiliation: Department of Veterinary Science, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido, Japan.
authors.forename: Taku
authors.initials: T
authors.lastname: Miyasho
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Taizo
authors.initials: T
authors.lastname: Hibi
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Abe
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Kitago
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Obara
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Itano
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Hiroya
authors.initials: H
authors.lastname: Takeuchi
authors.affiliation: Department of Pathology, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Michiie
authors.initials: M
authors.lastname: Sakamoto
authors.affiliation: Department of Surgery, Tokyo Medical and Dental University, Bunkyo, Tokyo, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Tanabe
authors.affiliation: Department of Laboratory and Vascular Medicine, Kagoshima University Graduate School of Medicine and Dental Sciences, Kagoshima, Kagoshima, Japan.
authors.forename: Ikuro
authors.initials: I
authors.lastname: Maruyama
authors.affiliation: Department of Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Kitagawa
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D024243:HMGB1 Protein / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D008099:Liver / Q000378:metabolism / Q000473:pathology; D017114:Liver Failure, Acute / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011897:Random Allocation; D017208:Rats, Wistar; D016019:Survival Analysis; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D024243:HMGB1 Protein; C104200:Hbp1 protein, rat; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha
keywords: Acute liver failure; Adenovirus vectors; Box-A; Gene delivery; High-mobility group box 1