Association Of CD4～+CD25～(high)T Cells And FOXP3 Gene Expression In Donor Grafts With Acute Graft-versus-host Disease Following HLA Identical Sibling Allogeneic Hematopoietic Stem Cell Transplantation

Objective To investigate the association of CD4⁺CD25^high T cells and FOXP3 gene expression in donor grafts with acute graft-versus-host disease after HLA-identical sibling allogeneic hematopoietic stem cell transplantation.Methods Twenty-four patients' donor grafts (peripheral blood or bone marrow) and their respective clinical characteristics were evaluated. Flow cytometry analysis was performed to assess the percentage of CD4⁺CD25⁺ and CD4⁺CD25^high T cells in cord blood, healthy controls' peripheral blood and donor grafts. The phenotype of magnetically sorted CD4⁺CD25⁺ T cells in donor grafts was also assayed with flow cytometry. Real-time relative quantitative reverse transcription -polymerase chain reaction was performed using ABI PRISM 7500 Sequence Detection System and SYBR Green I chemistry. Relative transcripts of FOXP3 mRNA were determined by the formula 1/ 2 ^{Ct foxp3-Ctβ2 -MG} with β2-MG as the internal control gene. The specificity of FOXP3 and β₂-MG amplifications was confirmed by analyzing the dissociation curves and electrophoresis of the target amplicon.Results The CD4+CD25+ T cells in cord blood were a...</sup>