Abnormalities of the Bone Marrow Immune Microenvironment in Patients with Prolonged Isolated Thrombocytopenia after Allogeneic Hematopoietic Stem Cell.

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Abnormalities of the Bone Marrow Immune Microenvironment in Patients with Prolonged Isolated Thrombocytopenia after Allogeneic Hematopoietic Stem Cell Transplantation Yang Song, Min-Min Shi, Yuan-Yuan Zhang, Xiao-Dong Mo, Yu Wang, Xiao-Hui Zhang, Lan-Ping Xu, Xiao-Jun Huang, Yuan Kong Biology of Blood and Marrow Transplantation Volume 23, Issue 6, Pages 906-912 (June 2017) DOI: 10.1016/j.bbmt.2017.02.021 Copyright © 2017 The American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 Percentage of each T cell subset in the BM microenvironment. (A and B) The percentage of BM CD3+CD4+ cells is lower (A) and the percentage of CD3+CD8+ cells is higher (B) in patients with PT compared with those with GGF. (C) The ratio of CD4+ cells to CD8+ cells is lower in patients with PT. (D and E) Percentages of effector T cells, naïve T cells, effector memory T cells, and central memory T cells among CD4+ T cells and CD8+ T cells. Statistical analyses were performed using the Mann-Whitney U test. The results are presented as percentage and are expressed as mean ± SD. P values < .05 were considered statistically significant. Biology of Blood and Marrow Transplantation 2017 23, 906-912DOI: (10.1016/j.bbmt.2017.02.021) Copyright © 2017 The American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 Immunophenotypic analysis of BM Th1, Th2, Th17, Treg, Tc1, and Tc2 cells. (A) The lymphocytic population was initially defined using forward scatter (FSC) and side scatter (SSC) gates. (B) The CD4+ T cells and CD8+ T cells were then gated based on the CD8 antigen. (C) Representative flow cytometry analyses of CD8−IFN-γ+ Th1 and CD8+IFN-γ+ Tc1 cells from a patient with PT, a patient with GGF, and an HD control. (D) CD8−IL-4+ Th2 and CD8+IL-4+ Tc2 cells from a patient with PT, a patient with GGF, and an HD control. (E) CD3+CD8−IL17A+ Th17 cells from a patient with PT, a patient with GGF, and an HD control. (F) CD3+CD8−CD25+Foxp3+ Treg cells from a patient with PT, a patient with GGF, and an HD control. Biology of Blood and Marrow Transplantation 2017 23, 906-912DOI: (10.1016/j.bbmt.2017.02.021) Copyright © 2017 The American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 Increased frequency of Th1, Tc1, and Th17 cells in the BM microenvironment of PT patients. Shown are the percentages of BM Th1 (A), Th2 (B), Th17 (G), and Treg (H) cells among CD4+ T cells and of Tc1 (D) and Tc2 (E) cells among CD8+ T cells. Th1/Th2 (C), Tc1/Tc2 (F), and Th17/Treg (I) ratios are indicated. Statistical analyses were performed using the Mann-Whitney U test. Biology of Blood and Marrow Transplantation 2017 23, 906-912DOI: (10.1016/j.bbmt.2017.02.021) Copyright © 2017 The American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 4 Secreted cytokine levels in BM plasma of PT patients, GGF patients and HD controls. IFN-γ (A), IL-4 (B), IL-17 (C), IL-6 (D), IL-21 (E), and TPO (F) levels in BM plasma were measured by cytometric bead assay and ELISA. The PT patients exhibited higher levels of IFN-γ and significantly higher levels of IL-17, IL-6, and IL-21. IL-4 and TPO levels in the plasma were similar in PT and GGF patients. Statistical analyses were performed using the Mann-Whitney U test. The results are presented as a percentage and are expressed as mean ± SD. Biology of Blood and Marrow Transplantation 2017 23, 906-912DOI: (10.1016/j.bbmt.2017.02.021) Copyright © 2017 The American Society for Blood and Marrow Transplantation Terms and Conditions