Diagnostic profiles for precision medicine in systemic sclerosis:stepping forward from single biomarkers towards pathophysiological panels

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Autoantibody profiles in systemic sclerosis: a comparison of diagnostic tests

Onderzoek naar de profilering van antilichamen in de prognose en classificatie van systemische sclerose. Het doel van het onderzoek was om commercieel beschikbare diagnostische assays voor de detectie van SSc-geassocieerde autoantilichamen te vergelijken. Daarnaast werder onderzoek gedaan naar de coëxistentie van meerdere SSc-geassocieerde autoantilichamen bij patiënten

Modulation of cytokine release by adalimumab.

<p>Peripheral blood mononuclear cells (PBMCs) from of eight healthy controls and 40 patients with hidradenitis suppurativa were stimulated for the production of (A) interleukin (IL)-1β with LPS of <i>Escherichia coli</i> O55:B5; (B) IL-6 with LPS of <i>E</i>.<i>coli</i> O55:B5; (C) IL-10 with LPS of <i>E</i>.<i>coli</i> O55:B5; (D) IL-10 with heat-killed <i>Candida albicans</i> (HKCA); (E) IL-10 with heat-killed <i>Staphylococcus aureus</i> (HKSA); (F) IL-1ra with HKSA; (G) IL-17 with HKCA; (H) IL-17 with HKSA. P values refer to comparisons of cytokine production without/with adalimumab; pNS: difference not-significant.</p

A proposed scheme of pathogenesis of HS.

<p>Patients with HS have monocytosis but cytokine responses of inflammatory monocytes and T17 lymphocytes are inhibited. This inhibition is mediated for T17 cells by tumour necrosis factor-alpha (TNFα). When cells migrate to tissues from the blood vessels through the vascular endothelium, capacity for excess production of TNFα, interleukin (IL)-1β, IL-1α, IL-6 and IL-17 is restored.</p

Changes of cytokine production and of C Reactive Protein with the clinical course of HS.

<p>PBMCs were isolated and serum was sampled from patients with Hurley III disease at baseline and after 8 weeks of treatment with etanercept. Patients are divided into those experiencing worsening of symptoms (n = 6) or clinical improvement (n = 7) at the end of treatment. No change was defined when disease flare-ups persisted with heavy purulence. Improvement was defined as decrease of Sartorius score more than 30% from the baseline. PBMCs were stimulated with LPS of <i>Escherichia coli</i> O55:B5; with heat-killed <i>Candida albicans</i> (HKCA); and with heat-<i>Staphylococcus aureus</i> (HKSA) for the production of (A) tumour necrosis factor-alpha (TNFα), (B) interleukin (IL)-1β, (C) IL-17 and (D) C Reactive Protein (CRP). P values refer to comparisons of change of cytokine production from the baseline between improved and worsened cases by the Mann-Whitney U test.</p

Clinical characteristics of patients enrolled in the study.

<p>Clinical characteristics of patients enrolled in the study.</p

Stimulation of IL-10 and IL-17 in relation with disease severity

<p>Production of IL-10 and IL-17 was stimulated in peripheral blood mononuclear cells (PBMCs) of patients with hidradenitis suppurativa (HS) by LPS of <i>Escherichia coli</i> O55:B5, heat-killed <i>Candida albicans</i> (HKCA) and heat-killed <i>Staphylococcus aureus</i> (HKSA). Patients are divided into those with Hurley I or Hurley II disease stage (n = 50) and into those with Hurley III disease stage (n = 40). P values refer to comparisons between disease stages by the Mann Whitney U test; pNS: difference not-significant.</p

Stimulation of circulating monocytes.

<p>Production of (A) tumour necrosis factor-alpha (TNFα), (B) interleukin (IL)-1β, (C) IL-6, (D) IL-10, (E) IL-10, (F) IL-22, and (G) IL-1ra by peripheral blood mononuclear cells (PBMCs) of eight healthy controls and of 80 patients with hidradenitis suppurativa (HS). PBMCs were stimulated with LPS of <i>Escherichia coli</i> O55:B5; with heat-killed <i>Candida albicans</i> (HKCA), and with heat-killed <i>Staphylococcus aureus</i> (HKSA). (H) Gene transcripts of <i>TNF</i> and of <i>IL-1</i> after stimulation with LPS. P values refer to comparisons with healthy controls by the Mann-Whitney U test; pNS: difference not-significant.</p

Cytokine concentrations in pus of patients with HS.

<p>(A) Concentrations of tumour necrosis factor-alpha (TNFα), interleukin (IL)-1β, IL-1α, IL-6, IL-10, IL-17 and Il-1ra were measured in the pus of the lesions of 28 patients of Hurley III disease stage. Lines represent medians. Correlations between (Β) the number of patrolling monocytes and pus TNFα; (C) the number of patrolling monocytes and pus IL-1β; and (D) IL-6 and years since disease onset. Classification patterns of patients according to pus cytokines is shown in panel E. Spearman’s coefficients of correlation and p values of significance are provided.</p

Nailfold capillaroscopy and candidate-biomarker levels in systemic sclerosis-associated pulmonary hypertension:A cross-sectional study

Objectives: Pulmonary hypertension is one of the leading causes of death in systemic sclerosis. Early detection and treatment of pulmonary hypertension in systemic sclerosis is crucial. Nailfold capillaroscopy microscopy, vascular autoantibodies AT1R and ETAR, and several candidate-biomarkers have the potential to serve as noninvasive tools to identify systemic sclerosis patients at risk for developing pulmonary hypertension. Here, we explore the classifying potential of nailfold capillaroscopy microscopy characteristics and serum levels of selected candidate-biomarkers in a sample of systemic sclerosis patients with and without different forms of pulmonary hypertension.Methods: A total of 81 consecutive systemic sclerosis patients were included, 40 with systemic sclerosis pulmonary hypertension and 41 with no pulmonary hypertension. In each group, quantitative and qualitative nailfold capillaroscopy microscopy characteristics, vascular autoantibodies AT1R and ETAR, and serum levels of 24 soluble serum factors were determined. For evaluation of the nailfold capillaroscopy microscopy characteristics, linear regression analysis accounting for age, sex, and diffusing capacity of the lungs for carbon monoxide percentage predicted was used. Autoantibodies and soluble serum factor levels were compared using two-sample t test with equal variances.Results: No statistically significant differences were observed in quantitative or qualitative nailfold capillaroscopy microscopy characteristics, or vascular autoantibody ETAR and AT1R titer between systemic sclerosis-pulmonary hypertension and systemic sclerosis-no pulmonary hypertension. In contrast, several serum levels of soluble factors differed between groups: Endostatin, sVCAM, and VEGFD were increased, and CXCL4, sVEGFR2, and PDGF-AB/BB were decreased in systemic sclerosis-pulmonary hypertension. Random forest classification identified Endostatin and CXCL4 as the most predictive classifiers to distinguish systemic sclerosispulmonary hypertension from systemic sclerosis-no pulmonary hypertension.Conclusion: This study shows the potential for several soluble serum factors to distinguish systemic sclerosis-pulmonary hypertension from systemic sclerosis-no pulmonary hypertension. We found no classifying potential for qualitative or quantitative nailfold capillaroscopy microscopy characteristics, or vascular autoantibodies