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The pronounced Th17 profile in systemic sclerosis (SSc) together with intracellular expression of TGFbeta and IFNgamma distinguishes SSc phenotypes.

Radstake TR, van Bon L, Broen J, Hussiani A, Hesselstrand R, Wuttge DM, Deng Y, Simms R, Lubberts E, Lafyatis R - PLoS ONE (2009)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Rheumatology, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands. tradstake73@gmail.com

ABSTRACT

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.

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SSc patients have more and more activated CD4+ T cells compared to healthy controls.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).
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pone-0005903-g001: SSc patients have more and more activated CD4+ T cells compared to healthy controls.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).

Mentions: Previous reports have described an increased CD4/CD8 ratio in SSc patients compared with healthy controls, however, recent markers permit a more refined analysis of T cell phenotype [2], [30]. Since effector T cells, suggested to be involved in SSc, arise from the CD4+ T cells, we first investigated T cell phenotype and activation. Consistent with previous studies, healthy controls (n = 13) displayed a considerably lower CD4+/CD8+ ratio than that observed in SSc patients (P<0.0001, figure 1a). We next investigated whether CD4+ cells in SSc patients expressed T cell activation markers, CD69 and GITR. Indeed, SSc patients on average expressed significantly higher levels (P<0.0001) of CD69 on CD4+ effector T cells compared with those from healthy controls (figure 1b). In contrast, CD69 expression was increased on CD8+ cells only from those patients with the edcSSc, but not on CD8+ cells from patients with lcSSc or ldcSSc (P<0.001). The expression of GITR displayed a similar pattern being higher on CD4+ cells from all subgroups of SSc patients compared to controls (P<0.001). Highest expression was seen in patients with edcSSc, with progressively less expression seen in patients with ldcSSc and lcSSc, respectively (figure 1b, left panel). These observations prompted us to further study the expression of these activation markers on memory (CD45Ro) and naive T cells (CD45Ra), since CD45Ro cells were previously found to be the main producers of IL-17. Interestingly, the expression of CD69 was significantly higher on both CD45Ro and CD45Ra positive T cells in all SSc patients compared with healthy controls, suggesting that both these cell populations are activated in SSc (figure 1b right panel).


The pronounced Th17 profile in systemic sclerosis (SSc) together with intracellular expression of TGFbeta and IFNgamma distinguishes SSc phenotypes.

Radstake TR, van Bon L, Broen J, Hussiani A, Hesselstrand R, Wuttge DM, Deng Y, Simms R, Lubberts E, Lafyatis R - PLoS ONE (2009)

SSc patients have more and more activated CD4+ T cells compared to healthy controls.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).
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Related In: Results  -  Collection

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pone-0005903-g001: SSc patients have more and more activated CD4+ T cells compared to healthy controls.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).
Mentions: Previous reports have described an increased CD4/CD8 ratio in SSc patients compared with healthy controls, however, recent markers permit a more refined analysis of T cell phenotype [2], [30]. Since effector T cells, suggested to be involved in SSc, arise from the CD4+ T cells, we first investigated T cell phenotype and activation. Consistent with previous studies, healthy controls (n = 13) displayed a considerably lower CD4+/CD8+ ratio than that observed in SSc patients (P<0.0001, figure 1a). We next investigated whether CD4+ cells in SSc patients expressed T cell activation markers, CD69 and GITR. Indeed, SSc patients on average expressed significantly higher levels (P<0.0001) of CD69 on CD4+ effector T cells compared with those from healthy controls (figure 1b). In contrast, CD69 expression was increased on CD8+ cells only from those patients with the edcSSc, but not on CD8+ cells from patients with lcSSc or ldcSSc (P<0.001). The expression of GITR displayed a similar pattern being higher on CD4+ cells from all subgroups of SSc patients compared to controls (P<0.001). Highest expression was seen in patients with edcSSc, with progressively less expression seen in patients with ldcSSc and lcSSc, respectively (figure 1b, left panel). These observations prompted us to further study the expression of these activation markers on memory (CD45Ro) and naive T cells (CD45Ra), since CD45Ro cells were previously found to be the main producers of IL-17. Interestingly, the expression of CD69 was significantly higher on both CD45Ro and CD45Ra positive T cells in all SSc patients compared with healthy controls, suggesting that both these cell populations are activated in SSc (figure 1b right panel).

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Rheumatology, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands. tradstake73@gmail.com

ABSTRACT

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.

Show MeSH
Related in: MedlinePlus