title: Interferon-gamma and interleukin-10 messenger RNA are up-regulated after orthotopic liver transplantation in tolerant rats: evidence for cytokine-mediated immune dysregulation.
abstract: BACKGROUND
  Immune regulation requires antigen recognition, signaling, activation, secretion of cytokines, and effector function by lymphocytes. Although there is redundancy in the activation and function of the immune response, some cytokines simultaneously promote and suppress different pathways of immunity. In the experiments reported here we compare cytokine gene expression within liver allografts from tolerant rats with normal and isografted liver tissue. We also compare the secretion of interferon-gamma (IFN-gamma) in the supernatant from mixed lymphocyte cultures by using peripheral blood lymphocytes stimulated against donor antigen.
  
  
  METHODS
  Orthotopic liver transplantations were performed using the cuff technique without hepatic artery revascularization. Nonisotopic in situ hybridization (ISH) was used to detect and localize messenger RNA to specific cells within tissue. Antisense DNA probes were generated to interleukin-2 (IL-2), IL-4, IL-10, and IFN-gamma. One-way mixed lymphocyte cultures were set up against irradiated donor splenocytes, and the supernatant was collected to measure IFN-gamma by enzyme-linked immunosorbent assay.
  
  
  RESULTS
  Expression of IFN-gamma and IL-10 was up-regulated in tolerant animals versus normal or isografted liver (p = 0.0002 and 0.0001, IFN-gamma and IL-10, respectively). In situ hybridization localized the expression of messenger RNA predominantly to the cytoplasm of the hepatocytes. Levels of IFN-gamma were higher in the supernatant from proliferating peripheral lymphocytes against donor antigen from tolerant animals versus naive control animals.
  
  
  CONCLUSIONS
  Expression of IFN-gamma and IL-10 is up-regulated in hepatocytes from allograft tissue after orthotopic liver transplantation. We believe that the up-regulation of IL-10 cross-regulates the effector function of IFN-gamma and supports cytokine-mediated immune dysregulation, which may be a mechanism of tolerance after orthotopic liver transplantation in rats.
authors.affiliation: Department of Surgery, Stanford University, Calif., USA.
authors.forename: E J
authors.initials: EJ
authors.lastname: Alfrey
authors.forename: D
authors.initials: D
authors.lastname: Most
authors.forename: X
authors.initials: X
authors.lastname: Wang
authors.forename: L K
authors.initials: LK
authors.lastname: Lee
authors.forename: B
authors.initials: B
authors.lastname: Holm
authors.forename: N R
authors.initials: NR
authors.lastname: Krieger
authors.forename: R K
authors.initials: RK
authors.lastname: Sibley
authors.forename: P
authors.initials: P
authors.lastname: Huie
authors.forename: D C
authors.initials: DC
authors.lastname: Dafoe
mesh_terms: D000818:Animals; D001483:Base Sequence; D001773:Blood Cells / Q000378:metabolism; D002455:Cell Division; D016207:Cytokines / Q000235:genetics / Q000502:physiology; D007107:Immune System / Q000503:physiopathology; D007108:Immune Tolerance; D017403:In Situ Hybridization; D007371:Interferon-gamma / Q000235:genetics* / Q000378:metabolism; D016753:Interleukin-10 / Q000235:genetics*; D016031:Liver Transplantation; D008214:Lymphocytes / Q000378:metabolism; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D011917:Rats, Inbred Lew; D011920:Rats, Inbred WF; D017207:Rats, Sprague-Dawley; D014018:Tissue Distribution
chemical_list: D016207:Cytokines; D015335:Molecular Probes; D012333:RNA, Messenger; D016753:Interleukin-10; D007371:Interferon-gamma