title: Intracellular interferon-γ staining analysis of donor-specific T-cell responses in liver transplant recipients.
abstract: OBJECTIVE
  Biomarkers that accurately reflect, detect, and/or predict detrimental immune responses to grafts are important in organ transplantation. We established a new detection method for alloreactive T cells on the basis of intracellular staining for interferon (IFN)-γ, using CD40-activated B cells as stimulators, and assessed temporal changes in alloreactive T-cell frequencies in patients who received liver transplantation.
  
  
  METHODS
  Peripheral blood mononuclear cells and CD40-activated B cells were used as responder and stimulator cells, respectively. The responder cells were cultured with the stimulator cells for 7 days, restimulated for 5 hours, and flow cytometrically tested by intracellular staining for IFN-γ.
  
  
  RESULTS
  The relative postoperative-preoperative ratio of donor-specific CD8+ T cells in the nonrejection group was significantly lower than that in the rejection group and found to be <1 in most individuals of the group throughout the postoperative periods, indicating an induction of donor-specific suppression of the CD8+ T-cell responses. In contrast, such differences were not found in the donor-specific CD4+ T cells. These results suggest that the relative postoperative-preoperative ratio of the donor-specific CD8+ T cells is a good indicator of graft rejection.
  
  
  CONCLUSIONS
  We established a new flow cytometric method for the detection of alloreactive T cells by intracellular staining for IFN-γ, using CD40-activated B cells as stimulator cells. Using this system, we found that the relative postoperative-preoperative ratio of the donor-specific CD8+ T cells is a possible evaluative indicator of the risk for graft rejection.
authors.affiliation: Department of Hepatobiliary Pancreatic and Transplant Surgery, Mie University Graduate School of Medicine, Tsu, Japan. yokanami@yahoo.co.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Okanami
authors.forename: K
authors.initials: K
authors.lastname: Tsujimura
authors.forename: S
authors.initials: S
authors.lastname: Mizuno
authors.forename: M
authors.initials: M
authors.lastname: Tabata
authors.forename: S
authors.initials: S
authors.lastname: Isaji
authors.forename: Y
authors.initials: Y
authors.lastname: Akatsuka
authors.forename: K
authors.initials: K
authors.lastname: Kuzushima
authors.forename: T
authors.initials: T
authors.lastname: Takahashi
authors.forename: S
authors.initials: S
authors.lastname: Uemoto
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001402:B-Lymphocytes / Q000276:immunology; D015415:Biomarkers / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D019013:CD40 Antigens / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D005260:Female; D005434:Flow Cytometry; D006084:Graft Rejection / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism*; D007564:Japan; D016031:Liver Transplantation / Q000276:immunology*; D018655:Lymphocyte Count; D008297:Male; D008875:Middle Aged; D011237:Predictive Value of Tests; D018570:Risk Assessment; D012307:Risk Factors; D013997:Time Factors; D016896:Treatment Outcome
chemical_list: D015415:Biomarkers; D019013:CD40 Antigens; D007371:Interferon-gamma