Increased circulating levels and salivary gland expression of interleukin-18 in patients with Sjögren's syndrome: relationship with autoantibody production and lymphoid organization of the periductal inflammatory infiltrate

IL-18, an immunoregulatory and proinflammatory cytokine, has been shown to play an important pathogenic role in Th1-driven autoimmune disorders. In this study, we evaluated the circulating levels and salivary-gland expression of IL-18 in patients with Sjögren's syndrome (SS), a mainly Th1-mediated disease. IL-18 serum levels were measured by ELISA in 37 patients with primary SS, 42 with rheumatoid arthritis, and 21 normal controls. We demonstrated high IL-18 serum levels in SS, similar to those in rheumatoid arthritis patients and significantly higher than in controls (P < 0.01). In addition, IL-18 serum concentrations were significantly higher in anti-SSA/Ro(+ )and anti-SSB/La(+ )than in anti-SSA/Ro(- )and anti-SSB/La(- )SS patients (respectively, P = 0.01, P < 0.01). Serum IL-18 correlated strongly with anti-SSA/Ro (P = 0.004) and anti-SSB/La (P = 0.01) titers. Salivary gland IL-18 expression was investigated by single/double immunohistochemistry in 13 patients with primary SS and in 10 with chronic sialoadenitis, used as controls. The expression of IL-18 was also examined in periductal inflammatory foci in relation to the acquisition of features of secondary lymphoid organs such as T–B compartmentalization, formation of follicular dendritic cell networks, and presence of germinal-center-like structures. IL-18 expression in SS salivary glands was detected in 28 of 32 periductal foci of mononuclear cells (87.5%), while no IL-18 production by infiltrating cells was detected in patients with chronic sialoadenitis. Within the inflammatory foci, IL-18 immunoreactivity co-localized almost exclusively with CD68(+ )macrophages. In addition, IL-18 was found in 15 of 19 foci (78.9%) with no evidence of T–B cell compartmentalization (nonsegregated) but in 100% of the segregated aggregates, both in T- and B-cell-rich areas. Strikingly, IL-18 was strongly expressed by CD68(+ )tingible body macrophages in germinal-centre-like structures both in SS salivary glands and in normal lymph nodes. IL-18 expression was observed in the ducts of all SS biopsies but in only 4 of 10 patients with nonspecific chronic sialoadenitis (P < 0.01). This study provides the first evidence of increased circulating levels and salivary gland expression of IL-18 in SS, suggesting an important contribution of this cytokine to the modulation of immune inflammatory pathways in this condition

In-hospital percentage BNP reduction is highly predictive for adverse events in patients admitted for acute heart failure: the Italian RED Study

Introduction: Our aim was to evaluate the role of B-type natriuretic peptide (BNP) percentage variations at 24 hours and at discharge compared to its value at admission in order to demonstrate its predictive value for outcomes in patients with acute decompensated heart failure (ADHF). Methods: This was a multicenter Italian (8 centers) observational study (Italian Research Emergency Department: RED). 287 patients with ADHF were studied through physical exams, lab tests, chest X Ray, electrocardiograms (ECGs) and BNP measurements, performed at admission, at 24 hours, and at discharge. Follow up was performed 180 days after hospital discharge. Logistic regression analysis was used to estimate odds ratios (OR) for the various subgroups created. For all comparisons, a P value 46% at discharge had an area under curve (AUC) of 0.70 (P 300 pg/mL. A BNP reduction of 25.9% after 24 hours had an AUC at ROC curve of 0.64 for predicting adverse events (P 46% was 4.775 (95% confidence interval (CI) 1.76 - 12.83, P 300 pg/mL and whose percentage decrease at discharge was 46% was 9.614 (CI 4.51 - 20.47, P 46% at hospital discharge compared to the admission levels coupled with a BNP absolute value < 300 pg/mL seems to be a very powerful negative prognostic value for future cardiovascular outcomes in patients hospitalized with ADHF

Thirty day post discharge BNP levels predict outcomes better than discharge levels - Italian RED study

Introduction: Heart failure (HF) patients often relapse into acute decompensation and consequent rehospitalization resulting in social and economic burdens. BNP has been previously shown to be prognostic at hospital discharge, but it has not been well characterized at time points after discharge from the emergency department (ED). Hypothesis: To determine the prognostic utility of BNP levels and changes in the emergency department (ED) and follow-up time-points. Methods: 276 HF patients were enrolled from 8 centers in Italy. The primary endpoints were cardiac associated rehospitalizations and deaths. Results: The Mean/Median BNP concentration at ED presentation was (mean±SE) 707.94±23.44pg/ml. Changes in BNP levels from presentation to 24h: –549.54±26.91pg/ml (p<0.001); to 72h: –426.57±26.91pg/ml (p<0.001); and at discharge: –323.59±29.51pg/ml (p<0.001). The mean/median BNP levels at follow-up time-points were 407.38±24.54 at 30 days, 363.07±29.51 at 60 days and 281.83±26.91pg/ml at 180 days. The BNP level at the 30 day follow-up was found to be highly predictive for events up to 180 days with an Area Under Curve (AUC) of 0.716 (p< 0.008) with a cut-point of 355pg/ml BNP and a hazard ratio of 8.56 (p = .014). The other considered BNP measurements did not significantly contribute to the prediction of the outcome. The discharge BNP level failed to significantly predict short term (within 30 days) outcomes (AUC: 0.647; p=0.054), and shows similar prognostic ability for long term (180 days) (AUC: 0.605; p<0.039).Conclusions: Tracking BNP levels post-discharge from the hospital may be equally important as the discharge BNP. Our results suggest that an increase in BNP at 30 days from the discharge BNP can be highly prognostic and may be a useful tool to discriminate between patients at higher risk for future events. This time period may be a crucial window to monitor BNP stability in the outpatient setting

IL-18 stimulates B-type natriuretic peptide synthesis by cardiomyocytes in vitro and its plasma levels correlate with B-type natriuretic peptide in non-overloaded acute heart failure patients

Background: An altered IL-18 pathway in heart failure (HF) has recently been described and this cytokine was shown tobe of clinical and prognostic utility. Cardiomyocytes are a target of this cytokine which exerts inflammatory, hypertrophic,and profibrotic activities. B-type natriuretic peptide is a cardiac hormone produced in response to cardiac filling to regulatecardiovascular homeostasis. The aim of the study was to verify the ability of IL-18 to induce B-type natriuretic peptidesynthesis in vitro and to analyse the relationship between these two molecules in plasma in vivo from acute HF patients.Methods and Results: We demonstrated the ability of IL-18 to directly stimulate a murine cardiomyocyte cell line toexpress the B-type natriuretic peptide gene, synthesize the relative protein through a PI3K-AKT-dependent transduction,and induce a cell secretory phenotype with B-type natriuretic peptide release. A correlation between IL-18 and B-typenatriuretic peptide plasma levels was found in non-overloaded acute HF patients, and in subgroups of acute HF patientswith diabetes and coronary artery disease. Acute HF patients with renal failure had significantly higher IL-18 plasma levelsthan patients without. IL-18 plasma levels were correlated with C-reactive protein plasma levels.Conclusions: This study provides the first evidence of the ability of IL-18 to induce B-type natriuretic peptide synthesisin vitro and outlines the relationship between the two molecules in acute HF patients with an ongoing inflammatory status

Degree of lymphoid organization of the periductal lymphocytic infiltrates in salivary gland of patients with Sjögren's syndrome (SS)

<p><b>Copyright information:</b></p><p>Taken from "Increased circulating levels and salivary gland expression of interleukin-18 in patients with Sjögren's syndrome: relationship with autoantibody production and lymphoid organization of the periductal inflammatory infiltrate"</p><p>Arthritis Research & Therapy 2004;6(5):R447-R456.</p><p>Published online 3 Aug 2004</p><p>PMCID:PMC546280.</p><p>Copyright © 2004 Bombardieri et al.; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.</p> Paraffin-embedded sections were double-stained for CD3 (brown) and CD20 (purple) (a–c) and single-stained with CD21 (d). Inflammatory foci were classified as nonsegregated when T and B lymphocytes were not compartmentalized in distinct areas (a), as segregated in the presence of evident compartmentalization of T and B cells (b), and as segregated with germinal-centre-like structures (arrow) when a clear histological appearance (c) and networks of CD21follicular dendritic cells (d) were observed. Original magnification × 200

Immunohistochemical (IHC) detection of IL-18 in salivary glands of patients with Sjögren's syndrome (SS) and in nonspecific chronic sialoadenitis

<p><b>Copyright information:</b></p><p>Taken from "Increased circulating levels and salivary gland expression of interleukin-18 in patients with Sjögren's syndrome: relationship with autoantibody production and lymphoid organization of the periductal inflammatory infiltrate"</p><p>Arthritis Research & Therapy 2004;6(5):R447-R456.</p><p>Published online 3 Aug 2004</p><p>PMCID:PMC546280.</p><p>Copyright © 2004 Bombardieri et al.; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.</p> (a,b) Paraffin-embedded section of glands in SS, showing high amounts of IL-18-expressing cells distributed in a scattered fashion within the periductal mononuclear infiltrate. (c) IL-18-positive cells were also observed surrounding acini (arrows) in proximity with the inflammatory aggregate. (d–f) Paraffin-embedded sections of glands from patients with nonspecific chronic sialoadenitis, demonstrating the absence of IL-18 expression in mononuclear cells in nonfocal periductal infiltrates. (g) Paraffin-embedded sections of glands from patients with SS, double-stained for CD68 (brown) and IL-18 (purple), showed exclusive co-localization of IL-18 expression in most of the CD68macrophages (arrows) within the periductal inflammatory infiltrates. (h) Macrophages expressing a large amount of IL-18 (arrows) were also observed surrounding acini in contiguity with a focal lymphocytic aggregate. (i, same sample as g) Conversely, CD68macrophages adjacent to a nonfocal infiltrate remained single-stained. Original magnification (a,b,d) × 100, (c,e–i) × 200

Relationship between IL-18 expression and B-/T-cell compartmentalization and germinal-center-like (GC-like) structures in the salivary glands of patient with Sjögren's syndrome (SS) and, for comparison, in normal lymph nodes

<p><b>Copyright information:</b></p><p>Taken from "Increased circulating levels and salivary gland expression of interleukin-18 in patients with Sjögren's syndrome: relationship with autoantibody production and lymphoid organization of the periductal inflammatory infiltrate"</p><p>Arthritis Research & Therapy 2004;6(5):R447-R456.</p><p>Published online 3 Aug 2004</p><p>PMCID:PMC546280.</p><p>Copyright © 2004 Bombardieri et al.; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.</p> Representative section of a large segregated aggregate double-stained for CD20 (brown) and IL-18 (purple) (a), and (b) sequential section with an irrelevant antibody replacing the anti-CD20, demonstrating the presence of IL-18-producing cells both in the T-cell (a, arrows) and B-cell (b, arrows) areas. (c) Single staining for IL-18, demonstrating a large number of IL-18-producing cells within ectopic GC-like structures in salivary gland from SS. (d) Double immunohistochemical staining for CD68 (brown) and IL-18 (purple), demonstrating the exclusive co-localization of IL-18 with CD68 macrophages. (e–h) An identical pattern of distribution in terms of IL-18 expression and co-localization with CD68 macrophages was observed in GCs of secondary lymphoid organs. Histomorphological analysis of the IL-18 positive cells within the GC showed evidence of engulfed apoptotic bodies in the cytoplasm (e) that identifies these cells as tingible body macrophages (TBMs). (f) Double immunohistochemical staining for CD68/IL-18 confirmed the exclusive co-localization of IL-18 with TBMs within the GC. Sequential sections in which the anti-CD68 (e), anti-IL-18 (g), or both the primary antibodies (h) were replaced with an isotype-matched irrelevant antibody confirmed the specificity of the double staining (h, negative control). Original magnification (a–d) × 200, (e-h) × 40

IL-18 expression in salivary gland ducts of patients with Sjögren's syndrome (SS) and nonspecific chronic sialoadenitis

<p><b>Copyright information:</b></p><p>Taken from "Increased circulating levels and salivary gland expression of interleukin-18 in patients with Sjögren's syndrome: relationship with autoantibody production and lymphoid organization of the periductal inflammatory infiltrate"</p><p>Arthritis Research & Therapy 2004;6(5):R447-R456.</p><p>Published online 3 Aug 2004</p><p>PMCID:PMC546280.</p><p>Copyright © 2004 Bombardieri et al.; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.</p> IL-18-positive ducts were detected in all the SS samples but in only a minority of those from chronic sialoadenitis. A considerable range of variability of IL-18 expression was observed in ducts among different samples. Within the same glandular lobule, positive and negative (arrowheads) adjacent ducts were observed (a). Ductal IL-18 expression was found in ducts surrounded (b) and not surrounded (c) by focal infiltrate, as well as in ducts characterized by periductal fibrosis (d). In contrast to ductal epithelial cells, no staining for IL-18 was found in acinar cells (a,d, stars). In a minority of patients with chronic sialoadenitis, we observed similar ductal staining patterns. Representative examples of positive (e) and negative (f) ductal IL-18 staining in different patients with chronic sialoadenitis are shown. Original magnification × 200