Endothelial Dysfunction and Pentraxin-3 in Clinically Stable Adult Asthma Patients.

BACKGROUND AND OBJECTIVE: Asthma is associated with low-grade systemic inflammation, prothrombotic state, and premature atherosclerosis. Objective: To evaluate the relationships between asthma, inflammatory biomarkers, and parameters of endothelial dysfunction. MATERIAL AND METHODS: We analyzed flow-mediated dilatation (FMD) of the brachial artery and intima-media thickness (IMT) of the common carotid artery using ultrasound in 92 clinically stable adult asthmatics and 62 well-matched controls. We also measured blood levels of selected inflammatory and asthma-specific biomarkers, including interleukin (IL) 4, IL-5, IL-6, IL-10, IL-12 (p70), IL-17A, IL-23, and interferon γ, as well as a disintegrin and metalloproteinase domain-containing protein 33 (ADAM-33). In addition, we assessed endothelial damage using 2 laboratory biomarkers: circulating von Willebrand factor (vWF) and pentraxin-3. We analyzed relationships between the study variables and asthma severity, lung function abnormalities, airway remodeling indices on computed tomography, and transthoracic echocardiography parameters. RESULTS: Asthmatics had higher IL-6, IL-10, and ADAM-33 levels. They were also characterized by 23% lower FMD% and 15% thicker IMT, as compared with controls (P<.001, both). In asthma, vWF was related to age (ß=0.28 [95%CI, 0.15-0.41]) and remained inversely associated with FEV1 (ß=-0.2 [95%CI, -0.05 to -0.35]). Surprisingly, a negative correlation was revealed between vWF and pentraxin-3 (ß=-0.17 [95%CI, -0.3 to -0.04]). Pentraxin-3 remained positively associated with airway remodeling indices. CONCLUSIONS: Asthma is characterized by endothelial dysfunction associated with airway obstruction. The biological role of pentraxin-3 is unknown, although our data suggest a protective role against endothelial damage and atherosclerosis.

Endothelial Dysfunction and Pentraxin-3 in Clinically Stable Adult Asthma Patients

Background: Asthma is associated with low-grade systemic inflammation, prothrombotic state, and premature atherosclerosis. Objective: To evaluate the relationships between asthma, inflammatory biomarkers, and parameters of endothelial dysfunction. Material and Methods: We analyzed flow-mediated dilatation (FMD) of the brachial artery and intima-media thickness (IMT) of the common carotid artery using ultrasound in 92 clinically stable adult asthmatics and 62 well-matched controls. We also measured blood levels of selected inflammatory and asthma-specific biomarkers, including interleukin (IL) 4, IL-5, IL-6, IL-10, IL-12 (p70), IL-17A, IL-23, and interferon γ, as well as a disintegrin and metalloproteinase domain–containing protein 33 (ADAM-33). In addition, we assessed endothelial damage using 2 laboratory biomarkers: circulating von Willebrand factor (vWF) and pentraxin-3. We analyzed relationships between the study variables and asthma severity, lung function abnormalities, airway remodeling indices on computed tomography, and transthoracic echocardiography parameters. Results: Asthmatics had higher IL-6, IL-10, and ADAM-33 levels. They were also characterized by 23% lower FMD% and 15% thicker IMT, as compared with controls (P<.001, both). In asthma, vWF was related to age (ß=0.28 [95%CI, 0.15-0.41]) and remained inversely associated with FEV1 (ß=–0.2 [95%CI, –0.05 to –0.35]). Surprisingly, a negative correlation was revealed between vWF and pentraxin-3 (ß=–0.17 [95%CI, –0.3 to –0.04]). Pentraxin-3 remained positively associated with airway remodeling indices. Conclusions: Asthma is characterized by endothelial dysfunction associated with airway obstruction. The biological role of pentraxin-3 is unknown, although our data suggest a protective role against endothelial damage and atherosclerosis (AU)

Antecedentes: El asma se asocia con inflamación sistémica de bajo grado, con un estado protrombótico y la existencia de aterosclerosisprematura.Objetivo: Evaluar las relaciones entre asma, biomarcadores inflamatorios y parámetros de disfunción endotelial.Material y métodos: Se ha analizado la dilatación mediada por flujo (DMF) de la arteria braquial y el grosor íntima-media (GIM) dela arteria carótida común mediante ecografía, en 92 adultos asmáticos clínicamente estables y 62 controles. También se midieron losniveles sanguíneos de determinados biomarcadores inflamatorios específicos de asma, incluyendo interleucina (IL) -4, IL-5, IL-6, IL-10,IL-12 (p70), IL-17A, IL-23, interferón γ, así como desintegrina y la metaloproteinasa que contiene el dominio proteína 33 (ADAM-33),junto con marcadores de laboratorio de daño endotelial: pentraxina-3 circulante y actividad plasmática del factor von Willebrand (vWF).Analizamos las relaciones de las variables estudiadas con la gravedad del asma, las anomalías de la función pulmonar, los índices detomografía computarizada (TC) pulmonar de remodelación de las vías respiratorias y los parámetros de ecocardiografía transtorácica.Resultados: Los asmáticos tuvieron mayores niveles de IL-6, IL-10 y ADAM-33. También se caracterizaron por tener un 23% menos deDMF y un 15% más grueso el GIM, en comparación con los controles (p <0,001, ambos). En el asma, vWF se relacionó con la edad (ß =0,28 [IC 95%: 0,15 a 0,41]) y se mantuvo en una relación inversa con FEV1 (ß = -0,2 [IC 95%: -0,05 a -0,35]). Sorprendentemente, seobservó una correlación negativa entre vWF y pentraxina-3 (ß = -0,17 [IC 95%: -0,3 a -0,04]). La pentraxin-3 se asoció positivamentecon los índices CT de remodelación de la vía aérea...(AU)

Facilitated expansion of Th17 cells in lupus nephritis patients.

The objective of this study was to investigate the mechanisms of T helper type 17 (Th17) expansion in lupus nephritis (LN) patients, and to determine whether or not it is associated with impaired function of regulatory T cells (Treg ). Major effector subsets of peripheral blood CD4+ T cells were assessed by flow cytometry in 33 LN patients with different activity of the disease and 19 healthy controls. The percentage of circulating Th17 cells was increased in LN (median = 1·2% of CD4+ compared to 0·6% in the control group, P < 0·01), while Treg cells remained unchanged (12·3 versus 12·1% in controls), resulting in a significantly lower Treg /Th17 ratio. Th17 expansion in the patient group was not related to LN activity, renal histology or blood and urine inflammatory biomarkers, but has been associated with a higher cumulative dose of cyclophosphamide. Treg cells in LN displayed mainly effector memory phenotype and expressed higher levels of transforming growth factor (TGF)-ß; however, their suppressant activity in lymphocyte proliferation assay was diminished compared to controls (~fourfold, P < 0·05). Co-culture of Treg and conventional CD4+ T cells resulted in marked suppression of the Th1 subset in both of the groups studied, but also in a potent expansion of Th17 cells, which in LN was twofold higher, as in controls (P < 0·05). In conclusion, our results demonstrate that Th17 expansion in LN is not increased during disease exacerbation, but is related to chronic immunosuppressive therapy. This immune signature is probably linked to the abnormal function of Treg cells, which were less suppressive in LN patients and even facilitated differentiation of Th17 cells.

Urinary cytokines and mRNA expression as biomarkers of disease activity in lupus nephritis.

Introduction Renal involvement is one of the most serious manifestations of systemic lupus erythematosus, but non-invasive assessment of inflammatory response in kidneys is challenging. In this study we aimed to validate markers of active lupus nephritis (LN) using urine immune profiling. Methods Urine and serum cytokines (17-plex array) and urine mRNA expression (∼40 immune and glomerular injury genes) were measured in LN patients with active disease ( n = 17) during remission ( n = 16) and in healthy subjects ( n = 18). Results Urine and serum levels of CCL2, CCL5 and CXCL10 were elevated in active LN as compared with disease remission (best discrimination for urine CXCL10 and CCL2) and correlated with LN activity. In the active disease, urinary cell transcriptome showed marked upregulation of proinflammatory cytokines (e.g. TNF, CCL2, CCL5, CXCL10), and type-1 immunity-related genes (e.g. CD3G, CD4, TBX21, IFNG). An active pattern of gene expression was also observed in four patients in remission, who had moderately increased urinary leucocyte count. Two patients from this group developed renal exacerbation during the following 3 months. Markers of type-17 immune axis (e.g. IL-17A) were not significantly increased in active LN. Conclusions Active LN patients were characterized by marked increase of proinflammatory mediators in the urine. Urine cytokines (CCL2 and CXCL10) and type-1 T-cell-related gene markers in the urine sediment had similar diagnostic performance in detection of active LN.

Asthma is associated with enhanced thrombin formation and impaired fibrinolysis.

BACKGROUND: There is evidence that altered blood coagulation and fibrinolysis are involved in the pathogenesis of asthma. Increased thromboembolic risk has been reported in asthmatics. OBJECTIVE: To investigate whether enhanced thrombin generation and impaired fibrinolysis occur in asthmatics. METHODS: Plasma thrombin generation profile together with a computational assessment of thrombin dynamics and fibrinolytic capacity expressed as clot lysis time (CLT) were determined in 164 consecutive patients with stable asthma and 72 controls matched for age, gender, weight and smoking. RESULTS: Asthma patients had 20.2% increased endogenous thrombin potential (ETP), 41.4% higher peak thrombin concentration, 61% higher maximal prothrombin conversion rate, 15.5% faster rate of thrombin formation (all, P < 0.0001) and 10% lower thrombin decay capacity (P = 0.0004) compared with controls. Asthmatics had also 14.4% longer CLT (P = 0.001) associated with 21.3% higher plasminogen activator inhibitor-1 (PAI-1) (P < 0.0001), and 13% higher plasma α2 -macroglobulin (P = 0.0002). Using ETP and CLT above 75th percentile of the control values as the cut-off levels, we found increased risks of enhanced thrombin generation and hypofibrinolysis in asthmatics, also after correction for potential confounders. ETP and CLT were associated inversely with forced expiratory volume in 1 s/vital capacity (FEV1 /VC) index, after adjustment for age and body mass index. Non-allergic asthma (n = 70, 42.6%) was characterized by 17.5% longer CLT (P = 0.02), which positively associated with PAI-1. Thrombin generation profile was not affected by allergy. CONCLUSION AND CLINICAL RELEVANCE: Asthma is associated with enhanced thrombin generation and impaired fibrinolysis, which might contribute to thromboembolic events in this disease.

Disseminated nocardiosis mimicking exacerbation of pulmonary sarcoidosis.

Nocardiosis is a rare, mixed suppurative and granulomatous, bacterial infection that can affect various organs, but most commonly lungs. Clinical manifestation is usually uncharacteristic; can mimic fungal, parasitic and mycobacterial infections or malignancy. Presentation can be also similar to that of the other granulomatous diseases, among them sarcoidosis. We present an unusual case of disseminated nocardiosis in a patient diagnosed before with sarcoidosis and treated with glucocorticoids. Clinical symptoms initially mimicked exacerbation of pulmonary sarcoidosis. The course of disease was severe.

Collagen receptors α(1)ß(1) and α(2)ß(1) integrins are involved in transmigration of peripheral blood eosinophils, but not mononuclear cells through human microvascular endothelial cells monolayer.

UNLABELLED: Asthma development may be driven by T helper lymphocytes with eosinophils playing the role of major effector cells. Recruitment of the inflammatory cells from blood to the airways is mediated by adhesive molecules, e.g. selectins and integrins. The most important in cell trafficking are integrins containing α(4) and ß(2) subunits. We hypothesized that also collagen receptors: α(1)ß(1) and α(2)ß(1), may be involved in cell migration to the inflammatory site in asthma. The aim of the study was to determine whether the inhibition of α(1)ß(1) or α(2)ß(1) integrins, affects transmigration of eosinophils and peripheral blood mononuclear cells (PBMC) through human microvascular endothelial cells monolayer (HMVEC) seeded on collagen IV coated wells in moderate persistent atopic asthmatics. METHODS: PBMC from 9 asthmatics were separated by gradient centrifugation followed by negative magnetic separation of eosinophils. Snake venom derived anti-adhesive proteins: viperistatin and VP12 (potent and selective inhibitors of α(1)ß(1) and α(2)ß(1) integrins, respectively) as well as VLO4 (a non-selective inhibitor of α(4)ß(1), α(5)ß(1) and α(v)ß(3) - used as a positive control), were used for inhibition studies. All anti-adhesive proteins studied, inhibited eosinophils, but only VLO4 affected PBMC transmigration through HMVEC. In bronchial asthma both collagen receptors α(1)ß(1) and α(2)ß(1) are likely to be involved in eosinophil transmigration to the inflammatory site. The role of α(2)ß(1) on lymphocytes is probably different. As the α(2)ß(1) integrin has been described as a stimulator of collagen accumulation, it might be, at least in part, responsible for asthma airway remodelling.

Central nervous system involvement as a major manifestation of rheumatoid arthritis.

A 58-year-old woman with an 8-year history of seropositive rheumatoid arthritis was admitted with right hemiparesis, history of seizures, fever, weight loss and headaches. Her blood tests revealed the presence of rheumatoid factor, elevated C-reactive protein and anti-cyclic citrullinated peptide antibodies (>200 RU/ml). Examination of cerebrospinal fluid demonstrated pleocytosis (118 cells/mm(3), predominantly lymphocytes) with elevated protein level (58 mg/dl); cultures were negative. Magnetic resonance imaging findings were suggestive for meningoencephalitis. Short course of high-dose corticosteroids and cyclophosphamide led to clinical improvement. Rheumatoid vasculitis was probably responsible for neurological symptoms.

Increased expression of collagen receptors: alpha1beta1 and alpha2beta1 integrins on blood eosinophils in bronchial asthma.

BACKGROUND: Eosinophils are one of the major effector cells in bronchial asthma. Their infiltration of airways correlates with the asthma severity. Recruitment and activation of eosinophils are partially mediated by integrins alpha4beta1 and alpha4beta7. Collagens type I and IV constitute important components of extracellular matrix and vascular basement membrane, respectively. Therefore, collagen-binding integrins (alpha1beta1 and alpha2beta1) may also play a role in eosinophil lung infiltration. OBJECTIVE: To evaluate the possible presence of alpha1beta1 and alpha2beta1 integrins on peripheral blood eosinophils from asthmatic subjects. METHODS: Collagen receptors were studied on eosinophils separated by immunomagnetic CD16-negative method from healthy donors (n=13) and patients with moderate persistent atopic bronchial asthma (n=15). Surface receptor identification was performed by flow cytometry and cell adhesion assay. RESULTS: Eosinophils isolated from the patients showed increased expression of both alpha1beta1 and alpha2beta1 integrins as compared with healthy controls. Moreover, adhesive function of eosinophils to collagen type IV was inhibited by snake venom disintegrins: viperistatin and obtustatin. These disintegrins contain KTS active motif and are specific inhibitors of alpha1beta1 integrin. CONCLUSION: We demonstrated for the first time that collagen receptors: alpha1beta1 and alpha2beta1 integrins are overexpressed on the surface of peripheral blood eosinophils of asthmatic subjects. Further studies may reveal potential application of KTS-disintegrins or their structural analogs for therapy of bronchial asthma.

Integrins in pulmonary inflammatory diseases.

Integrins are a family of heterodimeric transmembrane glycoproteins that mediate cell-cell and cell-matrix interactions. They participate in inflammatory reactions mainly by regulation of leukocyte migration, activation and survival. Elevated expression of the cell adhesion molecules, such as VCAM, ICAM and MAdCAM on the lumenal surface of vascular endothelial cells is a critical early event in organ inflammatory processes - including the lung. Adhesive interactions with their counter-receptors on leukocytes, selectins and integrins, result in migration of the leukocytes to the inflammed tissues. Integrins also participate in physiological and pathological reorganization of the lung structure during e.g. pneumonia healing, airway remodeling, angiogenesis, emphysema and pulmonary fibrosis. Agents that could inhibit the function of one or more of these integrins could provide a novel therapeutic strategy targeted to inhibit inflammatory and immune phenomena in the lung.

Hereditary hemorrhagic telangiectasia, factor V Leiden and antiphospholipid syndrome: a case report.

We report on a 57-year-old woman with three episodes of ischemic strokes and hereditary hemorrhagic telangiectasia (HHT). Tests for inherited and acquired thrombophilia showed elevated anticardiolipin immunoglobulin (Ig)M antibodies (on three separate occasions), anti-prothrombin IgG antibodies, and the heterozygous form of factor V Leiden. This is the first case of HHT, a primary antiphospholipid syndrome, combined with factor V Leiden. No detectable arteriovenous malformation was found and ischemic episodes, documented by computer tomography, were related to the presence of antiphospholipid antibodies and possibly the carriership of factor V Leiden mutation. Since aspirin provoked severe nasal hemorrhages, treatment with ticlopidine was initiated after the third stroke. Over an 18-month follow-up, ischemic episodes were absent and we regarded oral anticoagulation as unjustifiable.

Clinical and genetic features underlying the response of patients with bronchial asthma to treatment with a leukotriene receptor antagonist.

BACKGROUND: Treatment with antileukotriene drugs results in clinical improvement in many, though not all, patients with asthma. It can be hypothesized that the subpopulation of asthmatic patients, characterized by aspirin intolerance and cysteinyl-leukotriene overproduction, might profit most from antileukotriene treatment. MATERIALS AND METHODS: We compared the clinical response to montelukast in two well-matched groups of patients with mild asthma: 26 aspirin-intolerant asthmatics (AIAs) and 33 aspirin-tolerant asthmatics (ATAs). We also searched for possible predictors of the clinical response among the parameters reflecting the expression and production of cysteinyl-leukotrienes (cys-LTs). This was an 8-week, single-blind, placebo-controlled trial. RESULTS: Following a 3-week montelukast 10 mg day-1 treatment compared with placebo, there was a statistically significant reduction in the mean daytime and nocturnal asthma symptoms and beta 2-agonist use, as well as a significant improvement in the morning and evening peak expiratory flows and quality of life. Both groups showed a similar significant improvement in the parameters studied. Clinical response did not correlate with the baseline urinary LTE4 excretion level. Improvement of asthma was observed mostly in patients with a low baseline and non-IL-5 inducible expression of LTC4 synthase (LTC4S) mRNA in eosinophils. There was a trend toward a better response in carriers of LTC4S allele C, but no relationship to the CC10 genetic polymorphism. CONCLUSIONS: No difference in the clinical response to the montelukast treatment was observed between the AIAs and the ATAs.

[Clinical variability in two cases of systemic mastocytosis]. / Róznopostaciowosc kliniczna w dwóch przypadkach mastocytozy ukladowej.

Two cases of systemic mastocytosis with different clinical course were described. The first of them manifested with returning attacks of tachycardia with raise of blood pressure, followed by its drop and loss of consciousness. Abdominal pain and persistent diarrhea characterized the second one. The common signs in both cases were skin changes of urticaria pigmentosa, presence of Darier's symptom, very high blood levels of tryptase and prostaglandin D2 and rise in urinary LTE4 concentration. Differential diagnostic is discussed and histopathology bone marrow biopsies are presented.

Enhanced expression of the leukotriene C(4) synthase due to overactive transcription of an allelic variant associated with aspirin-intolerant asthma.

Aspirin-intolerant asthma (AIA), a distinct clinical syndrome affecting about 10% of adult asthmatics, appears to be unusually dependent on cysteine leukotriene (cys-LT) overproduction by pulmonary eosinophils. The gene coding for leukotriene (LT) C(4) synthase (LTC(4)S), the enzyme controlling cys-LT biosynthesis, exists as two common alleles distinguished by an A to C transversion at a site 444 nucleotides upstream of the translation start. We tested the hypothesis that this single nucleotide polymorphism (SNP) affects binding of transcription factors and influences the transcription rate, predisposing to AIA. Gel shift assay studies revealed that the (-444)C allele, conferring an activator protein-2 binding sequence, is an additional target for a transcription factor of histone H4 consensus. Introduction of the H4TF-2 decoy oligonucleotide into LTC(4)S-positive, differentiated HL-60 cells decreased accumulation of LTC(4) to 68%. Transfection of COS-7 with promoter construct increased expression of beta-galactosidase reporter for the (-444)C variant. The (-444)C allelic frequency was significantly higher in AIA patients (n = 76) as compared with matched aspirin-tolerant asthmatics (n = 110) and healthy controls (n = 75). Patients with AIA had also upregulated LTC(4)S messenger RNA expression in peripheral blood eosinophils. An inhaled provocation test with lysine-aspirin led to an increase in urinary output of LTE(4), which reached statistical significance only in carriers of the (-444)C allele. Our results suggest that a transcription factor, present in dividing and bone marrow resident progenitors of eosinophils, triggers LTC(4)S transcription in carriers of a common (-444)C allele due to binding with the histone H4 promoter element of the gene. Genetic predisposition to cys-LT pathway upregulation, a hallmark of AIA, can be related to overactive expression of the LTC(4)S (-444)C allele.

Thrombin generation measured ex vivo following microvascular injury is increased in SLE patients with antiphospholipid-protein antibodies.

Antiphospholipid-protein antibodies (APA) include lupus-type anticoagulant (LA) and antibodies recognizing complexes of anionic phospholipids (e.g. cardiolipin) and proteins (e.g. prothrombin and beta2-glycoprotein I). The presence of APA is associated with an increased risk of both arterial and venous thrombosis. However, the pathogenic mechanism leading to thrombosis in patients with APA remains unclear. We studied 32 patients with systemic lupus erythematosus (SLE) who were divided into two groups depending on the presence (n = 19) or absence (n = 13) of APA. Healthy volunteers (n = 12) matched by age and sex served as controls. In all subjects LA and IgG class anticardiolipin antibodies (ACA) were determined. Thrombin generation was monitored ex vivo measuring fibrinopeptide A (FPA) and prothrombin fragment F1 + 2 (F1 + 2) in blood emerging from a skin microvasculature injury, collected at 30 second intervals. In subjects with antiphospholipid antibodies mean FPA and F1 + 2 concentrations were significantly higher at most blood sampling times than in controls. In some SLE patients with APA the process of thrombin generation was clearly disturbed and very high concentrations of fibrinopeptide A were detected already in the first samples collected. Two minutes after skin incision SLE patients without APA produced slightly more FPA, but not F1 + 2, as compared to healthy subjects. Mathematical model applied to analyze the thrombin generation kinetics revealed that APA patients generated significantly greater amounts of thrombin than healthy controls (p = 0.02 for either marker). In contrast, in the same patients generation of thrombin in recalcified plasma in vitro was delayed pointing to the role of endothelium in the phenomenon studied. In summary, these data show for the first time that in SLE patients with antiphospholipid-protein antibodies thrombin generation after small blood vessel injury is markedly increased. Enhanced thrombin generation might explain thrombotic tendency observed in these patients.