8 research outputs found
The Pronounced Th17 Profile in Systemic Sclerosis (SSc) Together with Intracellular Expression of TGFβ and IFNγ Distinguishes SSc Phenotypes
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81194.pdf (publisher's version ) (Open Access)BACKGROUND: Systemic sclerosis (SSc) is an autoimmune disease where controversy on Th1/Th2 balance dominates. We investigated whether the recently discovered Th17 pattern was present in SSc. METHODOLOGY AND PRINCIPAL FINDINGS: Patients were subdivided as having limited cutaneous SSc (lcSSc, n = 12) or diffuse cutaneous SSc (dcSSc, n = 24). A further arbitrary subdivision was made between early dcSSc (n = 11) and late dcSSc (n = 13) based upon the duration of disease. As a comparator group 14 healthy controls were studied. CD3+ cells were isolated using FACS and subsequently studied for the expression of CD4, CD8, CD25, CD45Ro, CD45Ra, IL-23, GITR, CD69 and intracellular expression of IL-17, TGFbeta and IFNgamma using flow cytometry. Levels of IL-17, IL-6, IL-1alpha and IL-23 were measured using Bioplex assays. SSc patients had more and more activated CD4+ cells. In addition, CD4, CD45Ro and CD45Ra cells from all SSc patients highly expressed the IL23R, which was associated with a higher IL-17 expression as well. In contrast, IFNgamma and TGFbeta were selectively up regulated in SSc subsets. In line with these observation, circulating levels of IL-17 inducing cytokines IL-6, IL-23 and IL-1alpha were increased in all or subsets of SSc patients. CONCLUSION AND SIGNIFICANCE: The combination of IL-17, IFNgamma and TGFbeta levels in CD45Ro and CD45Ra cells from SSc patients is useful to distinguish between lSSc, ldSSc or edSSc. Blocking Th17 inducing cytokines such as IL-6 and IL-23 may provide a useful tool to intervene in the progression of SSc
The Pronounced Th17 Profile in Systemic Sclerosis (SSc) Together with Intracellular Expression of TGF beta and IFN gamma Distinguishes Different SSc Clinical Phenotypes
Cytokine expression pattern in SSc clinical phenotypes.
<p>Cytokine expression pattern in SSc clinical phenotypes.</p
SSc patients express high levels of IL-17-positive T cells and the co-presence of IL-17 with either IFNγ, IFNα or TGFβ reflects SSc phenotype.
<p>Panel A presents the percentage of IL-23R positive cells in the CD4+ positive pool of T cells (left panel) and CD45Ro and CD45Ra positive cells (right panel) from healthy controls (n = 14), and SSc patients with the lcSSc (n = 12), ldcSSc (n = 11) and edcSSc (n = 13) phenotype. Panel B shows the mean intracellular expression level of IL-17, IFNγ and TGFβ in CD4+ cells from each a representative individual from each tested group (left side). On the right side, the mean percentage of CD45Ro-positive or CD45Ra-positive cells that express IL-17, IFNγ or TGFβ and the mean intensity thereof are presented for the whole group of healthy controls (n = 14), and SSc patients with the lcSSc (n = 12), ldcSSc (n = 11) and edcSSc (n = 13) phenotype. Panel C depicts the percentage of CD25<sup>high</sup> cells that co-express IL-17 in healthy controls and different SSc phenotypes (* represents a p-value<0.01). Panel D represents the level of cytokines (IL-17 and IFNγ) spontaneously secreted by CD3+ T cells from healthy controls and SSc patients after 24 hrs of incubation. * <i>P</i>-value<0.0001, ** <i>P</i>-value<0.002, *** <i>P</i>-value<0.004.</p
SSc patients have more and more activated CD4+ T cells compared to healthy controls.
<p>Panel A depicts the percentage of CD4+ and CD8+ cells in the whole T cell pool (CD3+ cells) from healthy controls (n = 14), and SSc patients with the lcSSc (n = 12), ldcSSc (n = 11) and edcSSc (n = 13) phenotype. The CD3+ cells were isolated using MACS bead isolation after which CD4 and CD8 positivity was analyzed using flow cytometry. Panel B (left) depicts the percentage of CD4+ and CD8+ cells that were double-positive for the T cell activation markers CD69 and GITR. For this aim a representative individual from each group was selected. In the right panel, the percentage of CD4-CD69, CD8-CD69, CD4-GITR, CD8-GITR, CD45Ro-CD69 and CD45Ra-CD69 double positive cells is presented over the whole group of healthy donors (n = 14) and/or SSc patients (n = 36).</p
Increased level of Th17 inducing cytokines in the circulation of SSc patients.
<p>Panel A depicts the presence of IL-17 in the circulation of SSc patients and healthy controls. Panel B, C and D represents the levels of IL-6, IL-1α and IL-23, respectively. The circulating levels of IL-17, IL-1α and IL-23 was measured by ELISA whereas IL-6 was studied by Bioplex assays.</p