Studying Blood Samples in Young Patients With Cytopenia After a Donor Stem Cell Transplant

• Number of patients with complete chimerism as measured by standard short tandem nucleotide polymorphism PCR in whole blood and the different cell populations [ Designated as safety issue: No ]
  
• Number of patients with complete chimerism as measured by single nucleotide polymorphisms PCR in the different cell populations [ Designated as safety issue: No ]
  

• Number of patients with mixed chimerism and full hematological recovery at day 100 [ Designated as safety issue: No ]
  
• Number of patients with mixed chimerism and acute or chronic graft-versus-host disease [ Designated as safety issue: No ]
  

OBJECTIVES:

Primary

• To study hematopoietic chimerism in whole blood and different cell populations (i.e., CD14, CD15, CD 56, CD3, and CD19) as well as in dendritic cells and regulatory T cells after allogeneic hematopoietic stem cell transplantation with reduced intensity conditioning in patients with refractory cytopenia.
• To compare the results of chimerism obtained with standard short tandem nucleotide polymorphism PCR (sensitivity 1%) with those obtained with single nucleotide polymorphisms PCR (sensitivity 0.1- 0.01%).

Secondary

• To evaluate the relationship between mixed chimerism and hematological engraftment, overall survival, and event-free survival.
• To study the impact of mixed chimerism in plasmacytoid dendritic and regulatory T cells on the incidence of acute and chronic graft-versus-host-disease.

OUTLINE: This is a multicenter study.

Peripheral blood is collected from patients and donors prior to hematopoietic stem cell transplantation (HSCT). Patients also undergo blood sample collection on days 30, 60, 100, and 180 after transplantation. Peripheral blood cells are enriched and separated into lineage-specific subpopulations (i.e., CD3, CD14, CD15, CD19, and CD56) which are then divided equally for either DNA isolation via PCR or for flow cytometry. DNA concentrations in pre-HSCT donor and patient samples and in post-HSCT subpopulation samples are determined using quantitative real-time PCR. Samples are also analyzed for quantification of chimerism and detection of genetic markers via short tandem repeats- and sequence nucleotide polymorphism-based chimerism analyses.