title: Persistence of stable intragraft cell chimerism in rat liver allografts after drug-induced tolerance.
abstract: BACKGROUND
  Drug-induced tolerance of rat liver allografts is well documented. We analyzed cellular events during immunosuppressive therapy on day (d) 10 and in the late phase (d 100) after transplantation to assess for characteristics in the intrahepatic leukocyte (IHL) population in the phase of tolerance.
  
  
  METHODS
  Lewis rats served as recipients of Dark Agouti rat livers. Temporary immunosuppression with either cyclosporine (CsA) monotherapy (3 mg/kg/d) or triple therapy that consisted of a subtherapeutic CsA dosage (0.25 mg/kg/d) and monoclonal antibodies directed against the interleukin-2 receptor (IL-2R, CD25) and the intercellular adhesion molecule-1 (ICAM-1, CD54) was administered from postoperative d 0 to d 13. Cell migration and cell activation within liver grafts was assessed by standard histology and flow cytometry. IHL apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL).
  
  
  RESULTS
  Both CsA monotherapy and triple therapy prolonged liver allograft survival to more than 100 d and led to the induction of donor-specific tolerance. Untreated recipients rejected their allografts within 14 d. In both groups, donor-specific IHLs initially dropped to 18% to 25% on d 10, but they rebounded to as much as 40% on d 100 as a common characteristic of both groups. Within this population, donor-specific T cells were dominant. In both groups, increased numbers of activated (IL-2R+) CD8+ T lymphocytes were present on d 100. No accumulation of apoptotic IHL was observed on d 100. Their proportion was unchanged in the triple therapy group and slightly decreased in the CsA group compared to the syngeneic controls.
  
  
  CONCLUSIONS
  The present study reveals that tolerant liver allografts are repopulated by donor-specific T lymphocytes. This phenomenon is independent of the type of applied immunosuppression. The persistence of activated CD8+ T cells in the phase of proven donor-specific tolerance on d 100 indicates that liver tolerance is associated with the state of a permanent intragraft immune activation. It seems that the coexistence of donor cells with infiltrating recipient cells within liver grafts, termed intrahepatic cell chimerism, is characteristic for tolerated liver allografts.
authors.affiliation: Experimental Transplantation Immunology of the Department of Surgery, Institute of Pathology, University of Wuerzburg, D-97060 Wuerzburg, Germany.
authors.forename: H J
authors.initials: HJ
authors.lastname: Gassel
authors.forename: C
authors.initials: C
authors.lastname: Otto
authors.forename: I
authors.initials: I
authors.lastname: Klein
authors.forename: U
authors.initials: U
authors.lastname: Steger
authors.forename: D
authors.initials: D
authors.lastname: Meyer
authors.forename: A M
authors.initials: AM
authors.lastname: Gassel
authors.forename: W
authors.initials: W
authors.lastname: Timmermann
authors.forename: K
authors.initials: K
authors.lastname: Ulrichs
authors.forename: A
authors.initials: A
authors.lastname: Thiede
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D018414:CD8-Positive T-Lymphocytes / Q000473:pathology; D002678:Chimera; D016572:Cyclosporine / Q000627:therapeutic use*; D004359:Drug Therapy, Combination; D006085:Graft Survival / Q000187:drug effects; D007108:Immune Tolerance; D007958:Leukocyte Count; D007962:Leukocytes / Q000473:pathology / Q000502:physiology; D008099:Liver / Q000473:pathology* / Q000503:physiopathology; D016031:Liver Transplantation / Q000276:immunology*; D008297:Male; D051381:Rats; D011914:Rats, Inbred BN; D011917:Rats, Inbred Lew; D011919:Rats, Inbred Strains; D013601:T-Lymphocytes / Q000473:pathology; D013997:Time Factors; D014019:Tissue Donors; D014184:Transplantation, Homologous
chemical_list: D000911:Antibodies, Monoclonal; D016572:Cyclosporine