title: The protective effect of intrasplenic transplantation of Ad-IL-18BP/IL-4 gene-modified fetal hepatocytes on ConA-induced hepatitis in mice.
abstract: BACKGROUND
  Concanavalin A (ConA)-induced hepatitis is an experimental murine model mirroring the pathology of human autoimmune hepatitis.
  
  
  OBJECTIVE
  To investigate the effects of intrasplenically transplanted fetal hepatocytes (BNL.CL2) transfected with recombinant adenovirus vector expressing the IL-18 binding protein (IL-18BP) and IL-4 fusion protein on ConA-induced hepatitis in mice.
  
  
  METHODS
  Ad-IL-18BP/IL-4 was used to infect BNL.CL2 cells. IL-4 and IL-18BP fusion protein expression were detected by ELISA and Western blotting. BNL.CL2 cells infected with Ad-IL-18BP/IL-4 were intrasplenically transplanted into mice. After 10 days, mice were injected with ConA (15 mg/kg), and sacrificed 18 hours later. Liver injury was assessed by serum transaminase and liver histology. TNF-α, IL-18, IL-4, IL-10, IL-12p70 and monocyte-chemoattracting protein (MCP)-1 levels in serum and liver homogenates were detected by ELISA. Signaling molecules in liver homogenates were analyzed by Western blotting.
  
  
  RESULTS
  Ad-IL-18BP/IL-4 effectively expressed the IL-18BP/IL-4 fusion protein for more than 14 days in BNL.CL12 cells. Treatment of mice with Ad-IL-18BP/IL-4-BNL.CL2 before ConA injection significantly reduced the elevated plasma levels of transaminases compared with ConA control groups. TNF-α, IL-18, IL-12p70 and MCP-1 levels in serum and liver homogenates from mice transplanted with Ad-IL-18BP/IL-4-BNL.CL2 were lower and IL-4 and IL-10 levels were higher than control groups. Phosphorylation levels of NF-κB p65, AKT, p38 and JNK1/2 in liver homogenates were markedly suppressed by Ad-IL-18BP/IL-4.
  
  
  CONCLUSIONS
  Ad-IL-18BP/IL-4 was effectively transfected into mouse BNL.CL2 cells. Intrasplenic transplantation of Ad-IL-18BP/IL-4-BNL.CL12 cells alleviated the severity of inflammation in ConA-induced experimental hepatitis and provides a useful basis for the targeted gene therapy of liver disease.
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Xueting
authors.initials: X
authors.lastname: Shao
authors.forename: Yun
authors.initials: Y
authors.lastname: Qian
authors.forename: Chenhuai
authors.initials: C
authors.lastname: Xu
authors.forename: Bo
authors.initials: B
authors.lastname: Hong
authors.forename: Wanhong
authors.initials: W
authors.lastname: Xu
authors.forename: Ling
authors.initials: L
authors.lastname: Shen
authors.forename: Changzhong
authors.initials: C
authors.lastname: Jin
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Wu
authors.forename: Xiangmin
authors.initials: X
authors.lastname: Tong
authors.forename: Hangping
authors.initials: H
authors.lastname: Yao
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D017690:Cell Transplantation; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000235:genetics / Q000517:prevention & control / Q000601:surgery; D003208:Concanavalin A / Q000009:adverse effects*; D016207:Cytokines / Q000378:metabolism; D005333:Fetus / Q000166:cytology; D006505:Hepatitis / Q000209:etiology / Q000235:genetics / Q000517:prevention & control* / Q000601:surgery; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics*; D015847:Interleukin-4 / Q000235:genetics*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D013154:Spleen / Q000166:cytology
chemical_list: D016207:Cytokines; D036341:Intercellular Signaling Peptides and Proteins; C118370:interleukin-18 binding protein; D003208:Concanavalin A; D015847:Interleukin-4