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. Author manuscript; available in PMC: 2019 Oct 1.
Published in final edited form as: Am J Transplant. 2018 Apr 17;18(10):2544–2558. doi: 10.1111/ajt.14718

Figure 3. Exogenous steroid hormone and IL-6 are important factors in enhancing MDSC differentiation from BMCs.

Figure 3

Figure 3

(A) Cytokines levels in plasma were measured, and linear regression analysis was performed. The scatter plots demonstrate correlation between MDSC numbers and plasma IL-6 levels (left), TNF-α (middle) and GM-CSF (right). Plasma IL-6 levels (Y) (X; r = 0.6888, p = 0.0278, correlation: Y = 3.8703 + 0.2420 X, n = 14), but not TNF-α (p = 0.3638, n = 15) and GM-CSF (p = 0.1942, n = 12), were moderately correlated with MDSC numbers. (B) Correlations between administered doses of steroid [normalized value/body weight (kg), n = 72], tacrolimus trough levels (n = 55), days after transplantation (n = 72), and absolute numbers of MDSCs were investigated using multiple linear regression analysis. The Spearman’s rank correlation coefficient (r) and p values are indicated in each scatter plots, and the red lines show fitted linear lines. (C–E) The bar graph demonstrates the numbers of M-MDSCs (white bars), PMN-MDSCs (black bars), and e-MDSCs (striped bars). BMCs were cultured for 6 or 7 days. Representative data from two independent experiments are shown. Culture medium was supplemented with G-CSF and GM-CSF, IL-6, and/or MP (C), with G-CSF, GM-CSF, IL-6 and various concentrations of MP (D), or with G-CSF, GM-CSF, IL-6, MP, and various concentrations of mifepristone (E), as indicated under the X-axis of the bar graphs. (F–H) CFSE-labeled, CD33-depleted mononuclear cells from BMCs were cultured with a cocktail of irradiated allogeneic B cells and flow-sorted autologous BMC-derived MDSCs at the indicated T : MDSC ratios under the X-axis. Proliferation of CD3+CD4+ (F) and CD3+CD8+ (G) cells and IFN-γ production in the cultured medium (H) were measured. The graphs demonstrate % inhibition compared to data from co-culture without MDSCs. Representative data from two independent experiments are shown.