Additional file 1 of The serum lipid profiles in immune thrombocytopenia: Mendelian randomization analysis and a retrospective study

Supplementary Material

The response of Tc17 and Th17 cells to DEX in ITP patients.

<p>The percentages of Tc17 and Th17 cells had a decreasing tendency with the increase in DEX concentration in culture media.</p

Correlations between Tc17 and Th17 cells in ITP patients and controls.

<p>(a, b) No significant correlation was demonstrated in newly-diagnosed (r = −0.3150, P = 0.1762) or CR ITP patients (r = 0.0773, <i>P</i> = 0.7017); (c) A positive correlation was shown between Tc17 and Th17 cells in controls (r = 0.7239, <i>P</i><0.0001).</p

CD8 cells and CD4∶CD8 ratio in ITP patients and controls.

<p>(a) Significant difference of the CD4∶CD8 ratios between ITP patients (median, 1.42; range, 0.26–4.18) and control group (median, 1.90; range, 0.93–3.95; *<i>P</i> = 0.029). (b) Elevated percentage of CD8+ T cells in ITP patients (median, 29.34%; range, 9.78%–68.97%) compared with controls (median, 22.35%; range, 12.89%–32.3%; *<i>P</i> = 0.025).</p

The ratio of <i>RORC, IL-6, TNF-α, IL-23</i> mRNA in healthy controls and MDS patients.

<p>(A) The ratio of <i>RORC</i> mRNA in E-MDS patients compared with that of healthy controls or L-MDS was 4.7 (*<i>P</i> = 0.0007) or 3.3 (*<i>P</i> = 0.002), respectively. (B) The ratio of <i>IL-6</i> mRNA in L-MDS patients compared with that of healthy controls or E-MDS was 5.3 (*<i>P</i> = 0.0001) or 2.4 (*<i>P</i> = 0.037), respectively. (C) The ratio of <i>TNF-α</i> mRNA in L-MDS patients compared with that of healthy controls or E-MDS was 10.6 (*<i>P</i> = 0.002) or 3.5 (*<i>P</i> = 0.049), respectively. (D) <i>IL-23p19</i> mRNA expression level among E-MDS, L-MDS and healthy controls was comparable (P>0.05). Bars represent SD.</p

Circulating percentages of Th17 cells, Th1 cells, and Th22 cells in MDS.

<p>(A) Circulating percentages of Th17 (CD4⁺ IL-17⁺) cells from healthy controls, E-MDS and L-MDS. Significantly increased percentage of Th17 cells was found in E-MDS patients (median, 1.90%; range, 0.58–6.01%) compared to L-MDS (median, 1.16%; range, 0.15–1.86%) (*<i>P</i> = 0.002) or healthy controls (median, 1.01%; range, 0.55–1.69%) (*<i>P</i> = 0.002). (B) Circulating percentages of Th1 (CD4⁺IFNγ⁺) cells from healthy controls, E-MDS and L-MDS. There was no significant difference between E-MDS (median, 9.65%; range, 6.16–21.08%) patients and L-MDS (median, 8.41%; range, 2.59–15.23%) or healthy controls (median, 9.06%; range, 6.01–14.02%). (C) Circulating percentages of Th22 (CD4⁺ IL-22⁺ IL-17⁻IFNγ⁻ ) cells from healthy controls, E-MDS and L-MDS. Significantly elevated percentage of Th22 cells was found in L-MDS patients (1.77±0.84%) compared to E-MDS (1.27±0.50%) (*<i>P</i> = 0.03) and healthy controls (0.71±0.17%) (*<i>P</i><0.0001). Obviously increased percentage of Th22 cells was shown between E-MDS and healthy controls (*<i>P</i> = 0.002). (D) Circulating percentages of Th17 cells from healthy controls, bone marrow (BM) blasts <5% and blasts ≥5% patients. The circulating percentages of Th17 cells remained significantly higher in blasts <5% compared with healthy donors. (*<i>P</i> = 0.003).</p

Demographic and clinical characteristics of MDS patients.

<p>Abbreviations: BM, bone marrow; n, number; RCUD, refractory cytopenia with unilineage dysplasia; RARS, refractory anemia with ring sideroblasts; RCMD, refractory cytopenia with multilineage dysplasia; RAEB, refractory anemia with excess blasts; CMML, chronic myelo-monocytic leukemia.</p

The percentages of circulating Tc17 and Th17 cells in ITP patients and controls.

<p>Heparinized peripheral whole blood from all subjects was stimulated with phorbol myristate acetate, ionomycin, and monensin for 4 h, then stained with labeled antibodies as described in the Design and Methods section. (a) Lymphocytes were gated by flow cytometry in Gate 1. (b,d,f) CD3+ T subsets were gated by flow cytometry; the plots in intern box Gate 2 represent CD3+ T cells. (c, e, g) The percentages of circulating Tc17 and Th17 cells from ITP patients and controls; the percentage of positive cells is shown in each panel.</p

Circulating percentages of Th17, Th1 and Th22 cells in representative healthy controls, E-MDS and L-MDS patients.

<p>Heparinized peripheral whole blood from 37 MDS(E-MDS, n = 17; L-MDS, n = 20)patients and 20 healthy PB donors were stimulated with phorbol myristate acetate, ionomycin, and monensin for 4 h, and then stained with labeled antibodies for FACS analysis. (A) Lymphocytes were gated by flow cytometry. (B, C, D) Representative FACS dot plots of circulating Th17 (CD4⁺IL-17⁺) cells from healthy controls, E-MDS and L-MDS patients. (E, F, G) Representative FACS dot plots of circulating Th1 (CD4⁺IFNγ⁺) cells from healthy controls, E-MDS and L-MDS patients. (H, I, J) Representative FACS dot plots of circulating Th22 (CD4⁺IL-22⁺IL-17⁻IFNγ⁻ ) cells from healthy controls, E-MDS and L-MDS patients. Numbers in plots indicate relative percentages per quadrant.</p

Tc17 and Th17 cells, and the Th17∶Tc17 ratio in ITP patients and controls.

<p>(a) Significantly elevated percentage of Tc17 was found in newly-diagnosed ITP patients (median, 0.79%; range, 0.25–3.58%) compared to controls (median, 0.30%; range, 0.04–1.80%; ***<i>P</i><0.0001) and CR patients (median, 0.39%; range, 0.10–1.54%; **<i>P</i> = 0.0017). (b) Statistically elevated percentage of Th17 cells was found in newly-diagnosed ITP patients (median, 2.24%; range, 0.72–8.18%) compared to controls (median, 1.19%; range, 0.04–4.53%; **<i>P</i> = 0.0016; [similar to our previous study]). (c) The Th17∶Tc17 ratio in newly-diagnosed patients (median, 2.5850; range, 1.4221–3.0714) was 0.76-fold lower than controls (median, 3.3852; range, 1.0000–5.8571; ***<i>P</i><0.0001) and 0.70-fold lower than CR cases (median, 3.6771; range, 2.3896–5.5000; ***<i>P</i><0.0001).</p