Altered Phenotype of Circulating Dendritic Cells and Regulatory T Cells from Patients with Acute Myocarditis

Figures

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  Figure 1. Percentages of plasmacytoid dendritic cells and myeloid dendritic cells from myocarditis patients and controls. PBMC from myocarditis and control subjects were immunostained and analyzed by five-color flow cytometry. (a) Gating tree from negative lineage marker (LINneg), HLADR + CD11c+, and BDCA1+ (myeloid dendritic cells-mDCs), and from HLADR + CD11c− and BDCA4+ (plasmacytoid dendritic cells-pDCs). (b) Percentage of myeloid DCs (LINnegHLA − DR + CD11c+) and BDCA1+ from myocarditis and healthy volunteers. (c) Percentage of plasmacytoid DCs (LINnegHLA − DR + CD11c−) and BDCA4+ from myocarditis patients and controls. ∗p < 0.05, ∗∗p < 0.0.

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  Figure 2. Percentages of costimulatory molecules in plasmacytoid and myeloid dendritic cells. PBMC from myocarditis and controls were immunostained, and cells were analyzed using flow cytometry. (a) Gating tree from lineage negative (LINneg), HLADR + CD11c + DCs (mDCs) additionally labeled for CD40+, CD80+ or CD86+, and (LINneg), HLADR+ CD11c− (pDCs) additionally labeled for CD40+, CD80+, or CD86+. (b) Percentages (left) and MFI (right) of costimulatory molecules in myeloid DCs. (c) Percentages (left) and MFI (right) of costimulatory molecules in plasmacytoid dendritic cells. ∗p < 0.05, ∗∗p < 0.01.

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  Figure 3. Expression of ILT2 and ILT4 in DCs from myocarditis and control subjects. Multiparametric flow cytometry gating tree for lineage negative (LINneg), HLA − DR + CD11c+, BDCA1+ ILT2+ or ILT4+ cells (mDCs) and (LINneg), HLA − DR + CD11c−, BDCA4+, and ILT2+ or ILT4+ (pDCs). Gating tree for the analysis of the expression of ILT2 and ILT4 in myeloid dendritic cells and plasmacytoid dendritic cells in PBMC from a control subject (a) and (b) a myocarditis patient. Myeloid dendritic cells showed similar percentages of ILT2 and ILT4 positive DCs compared with healthy volunteers (Figures 4(b) and 4(d)). Interestingly, myeloid DCs from myocarditis patients displayed higher surface expression (MFI) of ILT4 compared to controls (p = 0.03, Figure 4(d)).

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  Figure 4. Expression of ILT2 and ILT4 in DCs from myocarditis and control subjects. Percentage of ILT2 (a) and ILT4 (c) in plasmacytoid DCs: LINnegHLADR+CD11c−BDCA4+ (right) and ILT2 and ILT4 expression on pDCs (MFI) (right). Percentage of ILT2 (b) and ILT4 (d) in myeloid DCs: LINneg, HLADR+CD11c + BDCA1+ (left) and expression of ILT2 and ILT4 mean fluorescence intensity on mDCs (MFI) (right). Data are shown as median and interquartile range. ∗p < 0.05, ∗∗p < 0.01.

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  Figure 5. Regulatory T cells from myocarditis patients and controls. PBMC from myocarditis and control subjects were immunostained, and then cells were analyzed by five-color flow cytometry. (a) Gating tree for CD4 + CD25 + FOXP3+ cells. (b) Percentage of FOXP3+ cells in Treg cells from myocarditis patients and controls. (c) Median fluorescence intensity (MFI) of FOXP3+ cells in CD4 + CD25+ lymphocytes from myocarditis patients and controls. ∗p < 0.05, ∗∗p < 0.01.

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  Figure 6. Cytokine levels in the plasma of patients with myocarditis and control subjects. The concentrations of the indicated cytokines were determined using flow cytometry in plasma from myocarditis patients and healthy subjects. ∗p < 0.05, ∗∗p < 0.01.