Real-Time PCR Analysis of Chimerism in T Cell Subsets as an Early Predictor of Graft-Versus-Host Disease Following Allogeneic Stem Cell Transplantation

Ann Transplant. 2015 Dec 3:20:720-8. doi: 10.12659/aot.894621.

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.

Material and 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.

Publication types

  • Multicenter Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acute Disease
  • Adolescent
  • Adult
  • Chimerism*
  • Chronic Disease
  • Cohort Studies
  • Female
  • Graft Rejection
  • Graft Survival
  • Graft vs Host Disease / diagnosis
  • Graft vs Host Disease / etiology*
  • Hematologic Neoplasms / mortality
  • Hematologic Neoplasms / pathology
  • Hematologic Neoplasms / surgery*
  • Humans
  • Linear Models
  • Male
  • Middle Aged
  • Poland
  • Predictive Value of Tests
  • Prognosis
  • Real-Time Polymerase Chain Reaction / methods*
  • Retrospective Studies
  • Risk Assessment
  • Statistics, Nonparametric
  • Stem Cell Transplantation / adverse effects*
  • Stem Cell Transplantation / methods
  • T-Lymphocyte Subsets / immunology
  • Transplantation Conditioning / methods
  • Transplantation, Homologous
  • Young Adult