High-dose bee venom exposure induces similar tolerogenic B-cell responses in allergic patients and healthy beekeepers.

BACKGROUND: The involvement of B cells in allergen tolerance induction remains largely unexplored. This study investigates the role of B cells in this process, by comparing B-cell responses in allergic patients before and during allergen immunotherapy (AIT) and naturally exposed healthy beekeepers before and during the beekeeping season. METHODS: Circulating B cells were characterized by flow cytometry. Phospholipase A2 (PLA)-specific B cells were identified using dual-color staining with fluorescently labeled PLA. Expression of regulatory B-cell-associated surface markers, interleukin-10, chemokine receptors, and immunoglobulin heavy-chain isotypes, was measured. Specific and total IgG1, IgG4, IgA, and IgE from plasma as well as culture supernatants of PLA-specific cells were measured by ELISA. RESULTS: Strikingly, similar responses were observed in allergic patients and beekeepers after venom exposure. Both groups showed increased frequencies of plasmablasts, PLA-specific memory B cells, and IL-10-secreting CD73- CD25+ CD71+ BR 1 cells. Phospholipase A2-specific IgG4-switched memory B cells expanded after bee venom exposure. Interestingly, PLA-specific B cells showed increased CCR5 expression after high-dose allergen exposure while CXCR4, CXCR5, CCR6, and CCR7 expression remained unaffected. CONCLUSIONS: This study provides the first detailed characterization of allergen-specific B cells before and after bee venom tolerance induction. The observed B-cell responses in both venom immunotherapy-treated patients and naturally exposed beekeepers suggest a similar functional immunoregulatory role for B cells in allergen tolerance in both groups. These findings can be investigated in other AIT models to determine their potential as biomarkers of early and successful AIT responses.

Asthma is associated with reduced fibrinolytic activity, abnormal clot architecture, and decreased clot retraction rate.

The aim of this study was to assess whether steroid-naïve asthma modulates hemostasis. We evaluated the clot retraction rate (CRR), fibrinolysis rate (FR), clot density (CD) (by confocal microscopy), plasma levels of plasminogen activator inhibitor (PAI-1), and factor XIII (FXIII), NO in exhaled breath (FENO ), spirometry (FEV1 ) and eosinophil count (EOS) in 36 patients with allergic, steroid-naïve asthma and in 34 healthy controls. We observed significantly (P < 0.001) reduced CRR, FR, and FEV1 and increased FENO , EOS, PAI-1, FXIII, and CD in patients with asthma compared with controls. In patients with asthma, FR negatively correlated with CD, FXIII, PAI-1, FENO , and EOS and positively with FEV1 . FXIII positively correlated with CD. Clot retraction rate negatively correlated with FENO and positively with FEV1 (all P < 0.001). These novel findings suggest that asthma itself is associated with decreased CRR and reduced fibrinolytic potential resulting from alterations in clot architecture and elevated levels of plasma FXIII and PAI-1.

The effect of inhaled corticosteroids on the concentration of soluble CD163 in induced sputum of allergic asthma patients.

BACKGROUND AND OBJECTIVE: CD163 is a monocyte/macrophage-specific molecule whose expression is induced by corticosteroids and IL-10. The aim of this study was to evaluate the concentration of soluble CD163 (sCD163) in the induced sputum of asthmatic patients before and after therapy with inhaled corticosteroids (ICSs). PATIENTS AND METHODS: The study was performed in 24 patients with mild allergic asthma (AAs) and 10 healthy controls (HCs). In 18 AAs, induced sputum and serum samples were obtained before ICS therapy (T0) and 7 days later (T7). In the 6 AAs not treated with ICSs the procedures were performed at To and T7. The concentration of sCD163 in sputum and serum samples was evaluated using ELISA. RESULTS: There was no significant difference in mean (SD) baseline serum sCD163 concentration between AAs (1030 [449] ng/mL) and HCs (930 [334.5] ng/mL, P = .530). However, at To the mean sputum sCD163 concentration was significantly greater in AAs (4.78 [3.34] ng/mL) than in HCs (1.8 [0.41] ng/mL, P =.009). Treatment with ICSs resulted in a significant increase in sCD163 concentration in sputum (P < .0001) but not in serum (P =.679). No change in sputum or serum sCD163 concentration was detected in AAs who were not treated with ICSs. The change in sputum sCD163 concentration inversely correlated with changes in sputum eosinophilia or exhaled nitric oxide concentration. CONCLUSIONS: ICS therapy leads to local upregulation of sCD163 expression, which in turn may participate in the anti-inflammatory effects of ICS therapy.

The effect of inhaled corticosteroids on the concentration of soluble CD163 in induced sputum of allergic asthma patients

Background and Objective: CD163 is a monocyte/macrophage-specific molecule whose expression is induced by corticosteroids and IL-10. The aim of this study was to evaluate the concentration of soluble CD163 (sCD163) in the induced sputum of asthmatic patients before and after therapy with inhaled corticosteroids (ICSs). Patients and Methods: The study was performed in 24 patients with mild allergic asthma (AAs) and 10 healthy controls (HCs). In 18 AAs, induced sputum and serum samples were obtained before ICS therapy (T0) and 7 days later (T7). In the 6 AAs not treated with ICSs the procedures were performed at T0 and T7. The concentration of sCD163 in sputum and serum samples was evaluated using ELISA. Results: There was no significant difference in mean (SD) baseline serum sCD163 concentration between AAs (1030 [449] ng/mL) and HCs (930 [334.5] ng/mL, P=.530). However, at T0 the mean sputum sCD163 concentration was significantly greater in AAs (4.78 [3.34] ng/mL) than in HCs (1.8 [0.41] ng/mL, P=.009). Treatment with ICSs resulted in a significant increase in sCD163 concentration in sputum (P<.0001) but not in serum (P=.679). No change in sputum or serum sCD163 concentration was detected in AAs who were not treated with ICSs. The change in sputum sCD163 concentration inversely correlated with changes in sputum eosinophilia or exhaled nitric oxide concentration. Conclusions: ICS therapy leads to local upregulation of sCD163 expression, which in turn may participate in the anti-inflammatory effects of ICS therapy (AU)

Antecedentes y Objetivo: El CD163 es una molécula específica de monocitos/macrófagos cuya expresión es inducida por los corticosteroides y la interleucina 10 (IL-10). El objetivo de este estudio fue evaluar la concentración de sCD163 en el esputo inducido de pacientes asmáticos, antes y después del tratamiento con corticosteroides inhalados (ICS). Pacientes y Métodos: El estudio se realizó en 24 pacientes con asma alérgica leve (AA) y 10 controles sanos (HC). En 18 AA, se obtuvieron muestras de esputo y suero antes (T0) y a los 7 días (T7) tras la introducción del tratamiento con ICS. Asimismo, en 6 AA no tratados con ICS se llevaron a cabo los mismos procedimientos en T0 y T7. Se evaluó la concentración de sCD163 en esputo y suero de las muestras mediante ELISA. Resultados: No hubo diferencia en la concentración sérica basal media de sCD163 entre AA (1.030± 449 ng/ml) y los HC (930 ± 334,5 ng/ml, p = 0,530). Sin embargo, en T0 la concentración media de sCD163 esputo fue significativamente mayor en los AA (4,78 ± 3,34 ng/ml) que en HC (1,8 ± 0,41 ng/ml, p = 0,009). El tratamiento con ICS dio lugar a un aumento significativo de la concentración en el esputo de sCD163 (p < 0,0001), pero no en el suero (p = 0,679). No se detectaron diferencias en las concentraciones de sCD163 ni en el suero ni en el esputo del grupo de AA que no fueron tratados con ICS. La concentración sCD163 en esputo se correlacionó inversamente con la eosinofilia en esputo y la concentración NO exhalado. Conclusiones: El tratamiento con ICS induce la expresión local de sCD163, que a su vez puede mediar en el mecanismo anti-inflamatorio de este tratamiento (AU)

Glucocorticoids upregulate decreased IL-7 receptor expression in asthmatic patients and simian immunodeficiency virus-infected non-human primates.

Signaling through interleukin-7 receptor (IL-7R) is essential for regulation of T-cell homeostasis and survival. Previously, we and others have found diminished IL-7R levels in simian immunodeficiency virus (SIV) - infected non-human primates and human immunodeficiency virus (HIV) - infected patients. To date, it remains unknown whether changes in IL-7R expression could also be linked to non-infectious inflammatory diseases such as asthma or anti-inflammatory drug use. Here, we investigated through flow cytometry the levels of IL-7R expression on CD4+ and CD4- T-cells in asthmatic patients in relation to disease severity, immune status and glucocorticoid (GC) use. In addition, we sought to evaluate the effects of in vivo and in vitro GC treatment on IL-7R expression in both asthmatic patients and SIV-infected non-human primates. We demonstrated that expression of IL-7R on peripheral blood CD4+ T-cells was significantly decreased in clinically stable GC-naive mild and moderate asthmatic patients. Accordingly, the development of asthmatic reaction following bronchial allergen challenge performed in sensitized subjects was associated with a significant drop in levels of IL-7R on circulating CD4+ T-cells. In contrast, CD4+ T-cells from both, mild and moderate, but not severe asthmatics, treated with inhaled GC displayed levels of IL-7R similar to that seen in healthy controls. We did not find significant differences with serum or sputum interleukin-7 levels among healthy controls and GC-naïve and GC-treated asthmatic patients. Furthermore, both in vitro GC treatment and short-term oral GC administration to asthmatic patients resulted in a significant enhancement of IL-7R. Similarly, we demonstrated that GC-stimulated T-cells from SIV-infected non-human primates up-regulated IL-7R expression. Accordingly, experimental short-term systemic in vivo administration of GC to SIV-infected macaques led to enhancement of IL-7R expression on circulating T-cells. Our data indicate that GC bear potential to recover diminished IL-7R expression, as well in asthma as in lentiviral infection.

Lower proportions of CD4+CD25(high) and CD4+FoxP3, but not CD4+CD25+CD127(low) FoxP3+ T cell levels in children with autoimmune thyroid diseases.

The essence of autoimmune thyroid disease (AITD) is loss of tolerance of own tissues caused by malfunction of T lymphocytes, which affects the production of antibodies reacting with particular cell structures and tissues. Foxp3(+) regulatory T cells (Tregs) take part in the regulation of immune response and play a leading role in developing immune tolerance through active suppression. The aim of the study was to estimate the expression of CD4+CD25(high), CD4+CD25+CD127(low)FoxP3(+) and CD4+ FoxP3 T cells in patients with Graves' disease (GD) (n = 24, median age 15.5 years), in patients with Hashimoto's thyroiditis (HT) (n = 30, median age 15 years) in comparison with sex- and age-matched healthy control subjects (n = 30, median age 15 years). Polychromatic flow cytometry using a FACSCalibur (BD Biosciences) cytometer was applied to delineate T regulatory cell populations. In untreated patients with Graves' disease and HT we observed a significant decrease in CD4+FoxP3 (p < 0.001, p < 0.01) and CD4+CD25(high) (p < 0.016, p < 0.048) T lymphocytes as compared to the healthy control subjects. After 6-12 months of L-thyroxine therapy in HT cases these phenotypes of Tregs were normalized, yet no such changes were observed during GD therapy. The analysis of CD4+CD25+CD127(low)FoxP3+ T cells in the peripheral blood revealed comparable percentages of these cells in patients with thyroid autoimmune diseases to the controls. We conclude that the reduction number of Tregs with CD4+CD25(high) and CD4+FoxP3 phenotype suggests their role in initiation and development of autoimmune process in thyroid disorders.

Allergen challenge differentially affects the number of circulating monocyte subsets.

Peripheral blood monocyte (PBM) subsets play different roles in inflammatory response and tissue remodelling. The aim of this study was to investigate how allergen challenge affects the number of circulating PBMs in Dermatophagoides pteronyssinus (Dp) allergic patients (Dp-APs). Among 34 Dp-APs challenged, in 22 patients significant bronchoconstriction was demonstrated [responders (Rs)], while in 12, only upper respiratory symptoms were seen [non-responders (NRs)]. Twelve healthy, non-atopic subjects were used as controls (HCs). Expression of CD14, CD16 and CCR4 was evaluated by flow cytometry on the whole-blood samples before (T(0) ), 6 h (T(6) ) and 24 h (T(24) ) after the challenge. Plasma concentrations of CCL2, CX3CL1 and CCL17 were evaluated using ELISA. At T(0) , the mean percentage of CD14++ CD16+ PBMs in Rs (35.4%; 95%CI 26.9-43.9%) was significantly greater than in HCs (14.6%; 95%CI 7.3-21.8%; P = 0.006) and in NRs (17.5%; 95%CI 9.6-25.4%; P = 0.001). The baseline number of CD14++ CD16+ PBMs correlated with airway hyper responsiveness (AHR) (r = -0.507; 95%CI -0.834 to -0.432, P < 0.001). At T(24) , the number of CD14++ CD16+ PBMs significantly decreased in Rs but not in NRs and the numbers inversely correlated with plasma CCL17 concentration. Changes in the number of circulating CD14++ CD16+ cells after Dp challenge correlated with AHR (r = 0.706, 95%CI 0.43-0.861; P < 0.001). In all subjects, the greatest expression of CCR4 was found on CD14++ CD16+ PBMs. Expansion of CD14++ CD16+ monocytes in the peripheral blood with subsequent mobilization of those cells after allergen challenge may facilitate the development of AHR in Dp-APs.

Airway inflammation and eotaxin in exhaled breath condensate of patients with severe persistent allergic asthma during omalizumab therapy.

PURPOSE: The central role of IgE in allergic inflammation in asthma has provided a rationale for the development of omalizumab, the humanized monoclonal anti-IgE antibody. The aim of the study was to determine the effect of omalizumab treatment on changes in airway inflammatory process and eotaxin in exhaled breath condensate in patients with persistent severe allergic asthma. MATERIAL AND METHODS: The study was performed on a group of 19 patients with severe persistent allergic asthma treated with conventional therapy (according to GINA 2006) and with or without omalizumab (9 vs 10 patients). Eotaxin in exhaled breath condensate, exhaled nitric oxide, blood eosinophil count and serum ECP were measured before and after 16 weeks of therapy. RESULTS: In the group treated with omalizumab, a statistically significant decrease in the concentrations of eotaxin in EBC, FENO, serum ECP, and blood eosinophil count after 16 weeks of treatment was observed. Statistically significant correlations were revealed between the decrease in eotaxin and the decrease in FENO, serum ECP and blood eosinophil count after omalizumab therapy. CONCLUSIONS: Downregulation of eotaxin expression in the airways through limitation of eosinophilic inflammation could be essential in the beneficial effect of anti-IgE therapy with omalizumab in asthma patients.

Eotaxin in exhaled breath condensate of allergic asthma patients with exercise-induced bronchoconstriction.

BACKGROUND: Eosinophils are the key inflammatory cells in asthma, and more and more evidence suggests their crucial role in exercise-induced bronchoconstriction (EIB). Eotaxin, as the most important chemotactic factor for eosinophils, plays an important role in the pathogenesis of asthma. OBJECTIVES: The aim of the study was to evaluate the changes in eotaxin levels in exhaled breath condensate (EBC) following intensive exercise in allergic asthmatics. METHODS: The study was performed in a group of 27 asthmatics (17 with EIB, 13 without EIB) and 9 healthy volunteers. Changes induced by intensive exercise in the concentrations of eotaxin in EBC during the 24 h after an exercise test were assessed. The possible correlations of these measurements with the results of other tests commonly associated with eosinophilic airway inflammation were also determined. RESULTS: In asthmatic patients with EIB, a statistically significant increase in eotaxin concentrations in EBC collected during the first 24 h after an exercise test - with maximal increase after 6 h - was revealed. A statistically significant correlation between the maximum increase in eotaxin concentrations in EBC after exercise, and an increase in either serum eosinophil cationic protein or F(ENO) 24 h after exercise in the group of asthmatics with EIB, was observed. CONCLUSIONS: Our results confirm connections between EIB and airway eosinophilic inflammation. The increase of eotaxin in asthmatic airways, by promoting the migration and activation of eosinophils, may play an important role in upregulation and sustaining of the airway inflammation observed in EIB in asthmatic patients.

Bronchial hyperresponsiveness and airway inflammation in patients with seasonal allergic rhinitis.

BACKGROUND: Allergic rhinitis is a chronic inflammatory disease induced by an immunoglobulin (Ig) E-mediated reaction after exposure to an allergen. Many patients with allergic rhinitis and no clinical evidence of asthma show a heightened response to histamine. OBJECTIVES: The aims of the study were to measure changes in markers of airway inflammation in patients with seasonal allergic rhinitis and estimate changes in bronchial reactivity before and during the pollen season. METHODS: The study sample comprised 22 patients sensitized to grass pollen and 10 healthy volunteers. Based on the results of the bronchial provocation test (BPT) during the pollen season, we divided patients into those with and without bronchial hyperresponsiveness (BHR). We determined changes in nitrite and pH in exhaled breath concentrate (EBC), fraction of exhaled nitric oxide (FE(NO)), blood eosinophil count, and BPT results before and during the pollen season. RESULTS: In allergic rhinitis patients with BHR, we observed an increase in EBC nitrite (5.44 [2.33] vs 8.57 [3.35] nmol/mL, P = .02) and FE(NO) (20.90 [13.68] vs. 43.40 [31.60] ppb, P = .02) and a decrease in EBC pH (7.07 [0.33] vs. 6.74 [0.28], P = .01) during the pollen season. In allergic rhinitis patients with BHR, the increase in BHR was negatively correlated with increased FE(NO) and EBC nitrite and positively correlated with a decrease in EBC pH during the pollen season. CONCLUSIONS: Our results revealed a relationship between increased BHR in patients with seasonal allergic rhinitis and changes in airway inflammation markers. EBC pH, EBC nitrite concentration, and FE(NO) could act as prognostic markers for identifying patients at risk of developing asthma.

RANTES in exhaled breath condensate of allergic asthma patients with exercise-induced bronchoconstriction.

BACKGROUND: The response of asthmatics to exercise differs from that of healthy subjects, and the mechanisms responsible for exercise-induced bronchoconstriction (EIB) remain to be elucidated. OBJECTIVES: The aim of this study was to evaluate changes in RANTES levels in exhaled breath condensate (EBC) following intensive exercise in allergic asthmatics. METHODS: The study was conducted in a group of 19 asthmatics (11 with EIB and 8 without EIB) and 7 healthy volunteers. Changes in the concentrations of RANTES in EBC induced during the 24 h after intensive exercise were determined. Moreover, these measurements were tested for possible correlations with the results of other tests commonly associated with asthma as well as with changes in airway inflammation after exercise. RESULTS: In contrast to asthmatic patients without EIB and healthy controls, in asthmatics with EIB RANTES concentrations were statistically significantly increased in EBC collected during the first 24 h after an exercise test. There was a statistically significant correlation between the maximum increase in RANTES concentrations in EBC after exercise and either baseline exhaled nitric oxide (F(ENO)) or bronchial hyperreactivity to histamine and an increase in serum eosinophil cationic protein or F(ENO) 24 h after exercise in the EIB asthmatics. CONCLUSIONS: The increase in RANTES in asthmatic airways, promoting the migration and activation of inflammatory cells including eosinophils, may play an important role in the upregulation of airway inflammation after EIB in asthmatic patients.

Oral glucocorticoid treatment decreases interleukin-10 receptor expression on peripheral blood leucocyte subsets.

Glucocorticoids (GCS) are capable of stimulating the secretion of interleukin (IL)-10 by leucocytes; however, the potential of GCS to modulate leucocyte susceptibility to IL-10-mediated actions has not yet been studied. In the current paper, we performed a detailed cross-sectional analysis of IL-10 receptor (IL-10R) expression by CD4(+) and CD8(+) T cells, natural killer (NK) cells, monocytes and neutrophils. Next, we analysed the effects of short-term oral GCS treatment on surface IL-10R expression by various leucocyte subpopulations in asthmatic patients. All leucocyte subsets studied presented with substantial levels of surface IL-10R. The highest levels of IL-10R were found on monocytes, predominantly with CD14(2+)CD16(+) and CD14(+)CD16(+) phenotypes, and on CD4(+)CD25(high) T cells. In contrast, levels of IL-10R on CD8(+) T cells, NK cells and neutrophils were significantly lower and similar to each other in intensity. GCS treatment resulted in a significant decrease of IL-10R expression on all analysed peripheral blood leucocyte subsets. Our data suggest that down-regulation of IL-10R could counterbalance the otherwise suppressive action of GCS.

Changes in RANTES and beta-thromboglobulin after intensive exercise in patients with allergic asthma.

BACKGROUND: There is increasing evidence that exercise-induced bronchoconstriction (EIB) is associated with eosinophilic airway inflammation. In the pathogenesis of EIB the role of chemokines - responsible for promoting the migration and activation of inflammatory cells - as well as blood platelets, a potential source of those chemokines, remains unclear. METHODS: The study was conducted in a group of 19 asthmatics (11 with EIB, 8 without EIB) and 8 healthy volunteers. Changes in the plasma concentrations of RANTES and beta-thromboglobulin (beta-TG) induced by intensive exercise were determined. Moreover, the possible correlation of these measurements with the results of other tests used in the diagnosis of asthma as well as laboratory tests commonly associated with asthma were investigated. RESULTS: A comparison of the concentrations of beta-TG in all groups studied at rest did not reveal any significant differences. In all groups studied, 30 min after exercise elevated beta-TG concentrations were observed; the most significant increase was revealed in asthmatics with EIB. The baseline concentrations of RANTES before exercise in both groups of asthmatics were significantly higher in comparison to the group of healthy volunteers. After exercise, in the group of patients with EIB, a significant increase in RANTES concentrations was observed. These changes correlated with an increase in other markers of airway inflammation 24 h after exercise. CONCLUSIONS: We suggest that platelet activation, resulting in elevated RANTES release, could be one of the factors responsible for the increase of airway inflammation observed in consequence of EIB in asthmatics.

Soluble CD40 ligand and soluble P-selectin in allergic asthma patients during exercise-induced bronchoconstriction.

BACKGROUND: The interactions between CD40 and its ligand, CD40L, control humoral and cell-mediated immune responses. CD40 ligation may promote asthma-associated inflammatory responses in the airways. Many reports confirm the inflammatory basis of exercise-induced bronchoconstriction (EIB) in asthmatics. METHODS: The study was conducted in a group of 19 asthmatic patients (11 with EIB, 8 without EIB) and 8 healthy volunteers. We analyzed the changes in plasma concentrations of soluble CD40 ligand (sCD40L) and soluble P-selectin (sP-selectin) induced by intensive exercise. We also studied possible correlations with the results of measurements commonly associated with asthmatic inflammation. RESULTS: The study revealed statistically significant higher baseline concentrations of sCD40L--but not sP-selectin--in the group of asthmatics with EIB than in those without. In the asthmatic patients with EIB, sCD40L and sP-selectin concentrations increased significantly 30 minutes after exercise and returned to baseline 24 hours after exercise. Baseline concentrations of sCD40L correlated with baseline sP-selectin or fractional exhaled nitric oxide concentration (FE(NO)), an increase in sP-selectin 30 minutes after exercise, and changes in FE(NO) or bronchial hyperresponsiveness 24 hours after exercise. A statistically significant correlation between an increase in sCD40L concentrations 30 minutes after exercise and an increase in FE(NO) 24 hours after exercise or baseline eosinophil cationic protein was observed. CONCLUSION: After exercise in the group of allergic asthmatics with EIB, upregulation of CD40L by increased expression of inflammatory molecules and improved sensitivity of CD40-responsive cell types to the effects of proinflammatory cytokines may play an important role in the increased airway inflammation observed after postexercise bronchoconstriction.

Analysis of -675 4 g/5 G SERPINE1 and C-159T CD14 polymorphisms in house dust mite-allergic asthma patients.

BACKGROUND: Experimental studies indicate that endogenous plasminogen activator inhibitor-1 (PAI-1, encoded by the gene SERPINE1) modulates the immune response to lipopolysaccharide (LPS). On the other hand, LPS induces PAI-1 secretion. Activation of individual cells by LPS is facilitated by CD14. The single nucleotide polymorphisms -675 4G/5G in SERPINE1 and C-159T in CD14 are major determinants of PAI-1 and CD14 expression, respectively. OBJECTIVE: To evaluate the frequency of the -675 4G/5G SERPINE1 and C-159T CD14 polymorphisms in house dust mite (HDM) allergic asthma patients. METHODS: The polymorphisms were evaluated in unrelated inhabitants of northeastern Poland, including 372 HDM-allergic asthmatic patients and 160 healthy nonatopic control subjects using polymerase chain reaction. RESULTS: Both the C allele of CD14 and the 4G allele of SERPINE1 were more frequently encountered in HDM-allergic asthmatic patients than in healthy control individuals. When the 5G/5G-TT/CT genotype was considered as a nonrisk genotype, all other genotypes were associated with asthma. The odds ratios ranged from 3.96 (95% confidence interval, 1.56-10.1) for the 5G/5G-CC genotype to 10.7 (95% confidence interval, 5.1-24.9) for the 4G/4G-CC genotype. Bronchial reactivity to histamine and total serum immunoglobulin (Ig) E levels were predominantly associated with the 4G/5G SERPINE1 variants, while bronchial reactivity to Dermatophagoides pteronyssinus and serum concentrations of specific IgE against D pteronyssinus were predominantly associated with the C/T CD14 variants. Patients with 4G/4G-CC genotype had the lowest forced expiratory volume in 1 second and the highest bronchial reactivity. CONCLUSION: The SERPINE1 and CD14 polymorphisms studied here are associated with different aspects of bronchial reactivity and IgE response. Our results indicate that PAl-1 and CD14 may interact to affect susceptibility to allergic asthma.

Plasmin system regulation in an ovalbumin-induced rat model of asthma.

BACKGROUND: So far studies showing the role of the plasmin system in airway remodelling have been conducted using in vitro models. The aim of the present study was to determine plasmin system regulation in an in vivo rat model of asthma. METHODS: Asthma in Wistar rats was induced by ovalbumin (OVA) sensitization followed by an OVA challenge (OVA/OVA, n = 6). Control groups were saline-sensitized challenged with OVA (VEH/OVA, n = 6) and OVA-sensitized challenged with saline (OVA/VEH, n = 6). Plasmin system components were determined in the plasma by ELISA. Plasminogen activator inhibitor-1 (PAI-1) was localized by an immunohistochemical reaction. RESULTS: Sensitization and challenge with OVA caused thickening of the airway wall, hypertrophy of smooth muscle cells, infiltration of inflammatory cells, subepithelial fibrosis, epithelial and endothelial lesions. Serum total IgE was significantly higher in OVA-sensitized rats as compared to VEH-sensitized control groups. Tissue plasminogen activator activity was significantly decreased in asthmatic animals (4.48 +/- 0.4 vs. 6.7 +/- 0.3 ng/ml for OVA/OVA and OVA/VEH; p < 0.05), and PAI-1 activity was statistically significantly higher in asthma rats (0.8 +/- 0.05 vs. 0.5 +/- 0.03 ng/ml for OVA/OVA vs. OVA/VEH; p < 0.05). alpha2-Antiplasmin was higher in rats receiving OVA sensitization than in those that were sham sensitized (p < 0.05). Immunohistochemical staining for PAI-1in the lungs of asthmatic animals showed very strong PAI-1 expression in lung inflammatory cells. CONCLUSIONS: We have demonstrated for the first time the existence of PAI-1 in lung inflammatory cells of rats with asthma. This finding was consistent with the superiority of plasmin system inhibition over activation in plasma.

RANTES in exhaled breath condensate of stable and unstable asthma patients.

RANTES has been implicated in the allergic inflammation of asthma by promoting the migration and activation of the inflammatory cells, including eosinophils. The study was undertaken to evaluate RANTES levels in the exhaled breath condensate (EBC) of asthmatics with different degrees of asthma severity. EBC was collected from 33 patients with allergic asthma (11 with steroid-naïve mild asthma, 10 with ICS-treated, stable mild-to-moderate asthma, 12 with ICS-treated unstable, severe asthma) and seven healthy volunteers. In the three groups of asthmatics, RANTES concentrations in EBC were significantly higher compared with healthy volunteers. RANTES levels were significantly higher in patients with unstable asthma than in the two groups with stable disease. We observed statistically significant correlations between the concentrations of RANTES in EBC and F(ENO) in the three studied groups of asthmatics; notably, the correlation between the parameters described above was strong positive in the group of unstable and steroid-naïve stable asthmatics. We also discovered a significantly positive correlation between RANTES in EBC and the serum ECP or blood eosinophil count in the groups of asthmatics with severe, unstable asthma and between RANTES and serum ECP in the group of steroid-naïve stable asthmatics. Measurements of RANTES in EBC may provide another useful diagnostic tool for detecting and monitoring inflammation in patients with asthma.

Endothelin-1 in exhaled breath condensate of stable and unstable asthma patients.

Endothelins are proinflammatory, profibrotic, broncho- and vasoconstrictive peptides, which play an important role in the development of airway inflammation and remodeling in asthma. The study was undertaken to evaluate the endothelin-1 (ET-1) levels in exhaled breath condensate (EBC) of asthmatics with different degree in asthma severity. EBC was collected from 31 patients with allergic asthma (11 with steroid-naïve mild asthma, 10 with ICS-treated, stable mild-to-moderate asthma, 10 with ICS-treated unstable, severe asthma) and 7 healthy volunteers. In the three groups of asthmatics, ET-1 concentrations in EBC were significantly higher than in healthy volunteers. ET-1 levels were significantly higher in patients with unstable asthma than in the two groups with stable disease. There was a significant correlation between ET-1 levels and FENO in the three groups of asthmatics and between ET-1 and blood eosinophil counts in the group of patients with unstable asthma. Measurements of ET-1 in EBC may provide another useful diagnostic tool for detecting and monitoring inflammation in patients with asthma.