title: Real-Time PCR Analysis of Chimerism in T Cell Subsets as an Early Predictor of Graft-Versus-Host Disease Following Allogeneic Stem Cell Transplantation.
abstract: BACKGROUND
  Graft-versus-host-disease (GvHD) is the major cause of morbidity and mortality after stem cell transplantation. The development of early prediction methods is therefore of importance. Our aim was to analyze the usefulness of early donor chimerism monitoring after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in T cells and in CD4+ and CD8+ (lineage chimerism) for GvHD prediction.
  
  
  METHODS
  Chimerism was analyzed in 76 consecutive adult patients using RQ-PCR TaqMan technology on DNA extracted from Pan T, CD4+, and CD8+ cell subsets on Day 5, 10, 15 and 30 after allo-HSCT.
  
  
  RESULTS
  The threshold of chimerism predictive for GvHD was the same for all tested cell subsets. In acute myeloid leukemia (AML) patients treated with myeloablative conditioning (MAC), the threshold predictive for acute graft versus host disease was 95% and 99% for Day 10 and Day 15, respectively. In patients treated with reduced intensity conditioning (RIC), the threshold predictive for chronic graft versus host disease was 98% on Day 10. The differences were statistically significant.
  
  
  CONCLUSIONS
  Chimerism analysis in T cell subsets by RQ-PCR on Day 10 and Day 15 after transplantation is useful for prediction of aGvHD (AML patients after MAC) and cGvHD (patients after RIC). However, there was no difference in the results between chimerism in the T cell subsets. Our RQ-PCR protocol was highly sensitive and proved effective for analysis of lineage chimerism.
authors.affiliation: Department of of Immunohematology and Transfusion Medicine, Institute of Hematology and Transfusion Medicine, Warsaw, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Guz
authors.affiliation: Department of Haemopoietic Stem Cell Transplantation, Institute of Hematology and Transfusion Medicine, Warsaw, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Nasiłowska
authors.affiliation: Department of Haemopoietic Stem Cell Transplantation, Institute of Hematology and Transfusion Medicine, Warsaw, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Tomaszewska
authors.affiliation: Department of of Immunohematology and Transfusion Medicine, Institute of Hematology and Transfusion Medicine, Warsaw, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Orzińska
authors.affiliation: Department of of Immunohematology and Transfusion Medicine, Institute of Hematology and Transfusion Medicine, Warsaw, Poland.
authors.forename: Justyna
authors.initials: J
authors.lastname: Smolarczyk-Wodzyńska
authors.affiliation: 0.
authors.forename: Magdalen
authors.initials: M
authors.lastname: Krzemienowska
authors.affiliation: Department of Haemopoietic Stem Cell Transplantation, Institute of Hematology and Transfusion Medicine, Warsaw, Poland.
authors.forename: Kazimierz
authors.initials: K
authors.lastname: Hałaburda
authors.affiliation: Department of Microbiology, Warsaw Medical University, Warsaw, Poland.
authors.forename: Maciej
authors.initials: M
authors.lastname: Przybylski
authors.affiliation: Department of Haematology, Oncology and Internal Diseases, Warsaw Medical University, Warsaw, Poland.
authors.forename: Wiesław Wiktor
authors.initials: WW
authors.lastname: Jędrzejczak
authors.affiliation: Department of Haemopoietic Stem Cell Transplantation, Institute of Hematology and Transfusion Medicine, Warsaw, Poland.
authors.forename: Bożena
authors.initials: B
authors.lastname: Mariańska
authors.affiliation: Department of of Immunohematology and Transfusion Medicine, Institute of Hematology and Transfusion Medicine, Warsaw, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Brojer
mesh_terms: D000208:Acute Disease; D000293:Adolescent; D000328:Adult; D046528:Chimerism; D002908:Chronic Disease; D015331:Cohort Studies; D005260:Female; D006084:Graft Rejection; D006085:Graft Survival; D006086:Graft vs Host Disease / Q000175:diagnosis / Q000209:etiology*; D019337:Hematologic Neoplasms / Q000401:mortality / Q000473:pathology / Q000601:surgery*; D006801:Humans; D016014:Linear Models; D008297:Male; D008875:Middle Aged; D011044:Poland; D011237:Predictive Value of Tests; D011379:Prognosis; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012189:Retrospective Studies; D018570:Risk Assessment; D018709:Statistics, Nonparametric; D033581:Stem Cell Transplantation / Q000009:adverse effects* / Q000379:methods; D016176:T-Lymphocyte Subsets / Q000276:immunology; D019172:Transplantation Conditioning / Q000379:methods; D014184:Transplantation, Homologous; D055815:Young Adult