Increased apoptosis of peripheral blood T cells following allogeneic hematopoietic cell transplantation by Ming-Tseh Lin, Li-Hui Tseng, Haydar Frangoul,

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Presentation transcript:

Increased apoptosis of peripheral blood T cells following allogeneic hematopoietic cell transplantation by Ming-Tseh Lin, Li-Hui Tseng, Haydar Frangoul, Ted Gooley, Ji Pei, Alexandre Barsoukov, Yoshiki Akatsuka, and John A. Hansen Blood Volume 95(12):3832-3839 June 15, 2000 ©2000 by American Society of Hematology

Ming-Tseh Lin et al. Blood 2000;95:3832-3839 ©2000 by American Society of Hematology

Ming-Tseh Lin et al. Blood 2000;95:3832-3839 ©2000 by American Society of Hematology

Apoptosis among CD3+ T cells Apoptosis among CD3+ T cells.PBMCs were obtained from a patient 21 days following transplantation, cultured for 24 hours, and stained with PE-labeled anti-CD3 mAb (panels A, C, and E) or with PE-labeled anti-CD3 mAb and FITC-labeled IgG1 isotype control ant... Apoptosis among CD3+ T cells.PBMCs were obtained from a patient 21 days following transplantation, cultured for 24 hours, and stained with PE-labeled anti-CD3 mAb (panels A, C, and E) or with PE-labeled anti-CD3 mAb and FITC-labeled IgG1 isotype control antibody (panels B, D, and F) followed by 7AAD. Less than 1% of the T cells were stained with FITC-labeled IgG1 isotype control antibody. T-cell apoptosis was measured by using FSC and the intensity of 7AAD fluorescence (panel E or F) of cells noted in the R1 gate (panel A or B) and R2 gate (panel C or D). Ming-Tseh Lin et al. Blood 2000;95:3832-3839 ©2000 by American Society of Hematology

Wright-Giemsa stain and TUNEL assay of PBMCs obtained from a patient 21 days following transplantation, incubated for 24 hours, and separated by sorting into CD3+/7AAD+and CD3+/7AAD− cells.(A) Cells in the CD3+/7AAD+ fraction showed reduction in cell volume... Wright-Giemsa stain and TUNEL assay of PBMCs obtained from a patient 21 days following transplantation, incubated for 24 hours, and separated by sorting into CD3+/7AAD+and CD3+/7AAD− cells.(A) Cells in the CD3+/7AAD+ fraction showed reduction in cell volume, blebbing of plasma membrane, chromatin condensation, and nuclear fragmentation (original magnification × 1000), (B) while cells in the CD3+/7AAD− fraction showed a typical morphological feature of lymphocytes. (C) Cells in the CD3+/7AAD+ fraction were brightly fluorescent, indicating incorporation of fluorescein-dUTP into fragmented DNA (original magnification × 400), (D) while cells in the CD3+/7AAD− fraction showed no significant fluorescence. Ming-Tseh Lin et al. Blood 2000;95:3832-3839 ©2000 by American Society of Hematology

Correlation between T-cell apoptosis following 24-hour culture and HLA-DR expression in T cells before culture in 36 patients studied 19-23 days after transplantation.Linear regression curve is shown (R2 = 42.8%,P < .001). Correlation between T-cell apoptosis following 24-hour culture and HLA-DR expression in T cells before culture in 36 patients studied 19-23 days after transplantation.Linear regression curve is shown (R2 = 42.8%,P < .001). Ming-Tseh Lin et al. Blood 2000;95:3832-3839 ©2000 by American Society of Hematology

Ming-Tseh Lin et al. Blood 2000;95:3832-3839 ©2000 by American Society of Hematology

Ming-Tseh Lin et al. Blood 2000;95:3832-3839 ©2000 by American Society of Hematology

Apoptosis of CD4+ and CD8+ T cells following 24-hour culture Apoptosis of CD4+ and CD8+ T cells following 24-hour culture.We studied 30 patients with grade II-IV acute GVHD 19-23 days after transplantation (open circle) and 8 patients with grade 0-I GVHD 20-22 days after transplantation (closed circle). Apoptosis of CD4+ and CD8+ T cells following 24-hour culture.We studied 30 patients with grade II-IV acute GVHD 19-23 days after transplantation (open circle) and 8 patients with grade 0-I GVHD 20-22 days after transplantation (closed circle). Ming-Tseh Lin et al. Blood 2000;95:3832-3839 ©2000 by American Society of Hematology

Ming-Tseh Lin et al. Blood 2000;95:3832-3839 ©2000 by American Society of Hematology

Ming-Tseh Lin et al. Blood 2000;95:3832-3839 ©2000 by American Society of Hematology