6 research outputs found
Suppressive function of circulating T regulatory cells.
<p><b>A</b>, Gating strategy for Treg sorting and typical results after sorting. <b>B</b>, Proliferative responses of Treg, effector responder T cells and Treg:Tresp co-cultures (ratio1/1) upon CD3+CD28 stimulation. Histograms are expressed as mean value +/− SD of cpm for healthy controls (HC ; n = 18), remission patients (RP ; n = 15), and active patients (PA ; n = 19). <b>C</b>, The percentage of suppression of responding cells was determined as 1−(proliferation of co-culture/proliferation of responder population alone)×100. Data are presented as boxplots with whiskers from minimum to maximum. <b>D</b>, A suppressive assay mixing Treg and effector T cells from and AP patient and a healthy volunteer was performed. Black bars represent suppression with HC Treg, and white bars with patient Treg. The results of one experiment out of three with similar results are shown.</p
Frequency, number and phenotype of Treg in patients vs. controls.
<p>Histograms represent: <b>A</b>, Proportion of CD4<sup>+</sup>CD25<sup>high</sup>CD127<sup>low/−</sup> T cells within CD4<sup>+</sup>CD3<sup>+</sup> T cells (as gated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0018734#pone-0018734-g002" target="_blank">figure 2A</a>) for HC (mean, 6.8%, <i>n</i> = 18), remission, (mean, 6.9%, <i>n</i> = 15), and acute (mean, 6.7%, <i>n</i> = 19) AAV patients. No significant difference was observed for percentage of Treg cells between the 3 groups. <b>B</b>, Absolute numbers of CD4<sup>+</sup>CD25<sup>high</sup>CD127<sup>low/−</sup> T cells in whole blood for HC (mean, 60.3 cells/µL), remission (mean, 47.5 cells/µL), and acute (mean, 40.1cells/µL) AAV patients. <b>C</b>, MFI of FOX-P3 in gated Treg and <b>D</b>, Mean percentage of CD39<sup>+</sup> cells within Treg. Data are presented as boxplots with whiskers from minimum to maximum.</p
Characterization and count of dendritic cells using flow cytometry.
<p><b>A</b>, Gating strategy for identification of DC subsets, and expression of CD86, CD62L and CCR7 (blue line) vs. isotype control (red line) on DC subset. <b>B</b>, Quantification of DC, myeloid DC and plasmacytoid DC using flow cytometry is shown for healthy controls (HC ; n = 18), remission phase (RP ; n = 15) and acute phase (AP ; n = 19) AAV patients. Data are presented as boxplots with whiskers from minimum to maximum. <b>C</b>, Left: the histogram shows an exemple of CD62 staining on pDC from a control subject (empty histogram) and an AAV patient in acute phase (grey histogram) as compared to isotype control (dotted line); Right: CD62L expression on pDC and mDC is shown as MFI of CDL62 – MFI of isotype control. Data are presented as boxplots with whiskers from minimum to maximum.</p
Characterization of Treg using flow cytometry in controls and patients.
<p><b>A</b>, Expression of CD25, CD127 and FOX-P3 in CD4<sup>+</sup> T cells. Representative dot plots for healthy controls (HC), AAV patients in remission phase (AAV-RP) and AAV patients in acute phase (AAV-AP) showing CD25 vs. CD127 and CD25 vs. FOXP3 expression in gated CD4<sup>+</sup>CD3<sup>+</sup> T cells are shown. The gate use to identify Treg in fresh blood is shown in each CD25 vs. CD127 dot plot. <b>B</b>. Representative histograms showing expression of FOX-P3 (bold line) vs. isotype control (dotted line) and CD39 (inset) in CD4<sup>+</sup>CD25<sup>high</sup>CD127 <sup>low</sup> cells as gated in A.</p
Concentration of IL-17 in co-culture supernatants.
<p>IL-17 was quantified in supernatant using a multiplex fluorescent bead immunoassay: the levels of IL-17 produced by Treg, responder T cells and co-culture were not statistically different between the 3 groups.</p