Tesis sobre el tema "Airway (Medicine) - Inflammation"

Measuring the level of exhaled NO (eNO) in the breath is a new method to monitor airway inflammation in asthma and may have a role in the management of asthma. The hypotheses were that eNO will reflect the degree of inflammation in chronic asthma, and will indicate how anti- inflammatory therapy should be altered to improve asthma control. Three studies were performed to test the hypotheses. A cross sectional study was performed to define the normal range of eNO and to compare this range with those who have asthma or atopy. The second study was observational, to compare the level of eNO during and after an exacerbation of asthma. The third study was an interventional study to evaluate eNO in management of paediatric asthma. In this latter study the level of eNO was measured to monitor airway inflammation in asthmatic children with the intention of adjusting antiinflammatory drugs (inhaled glucocorticosteroids) according to the level of eNO. These studies have shown that the mean level of eNO was significantly higher in asthmatic compared with normal subjects, but not significantly different when compared with atopic non-asthmatic subjects. eNO was correlated with the number of positive skin prick tests in atopic subjects whether asthmatic or nonasthmatic. The eNO level was increased during acute exacerbations of asthma and decreased after two weeks with therapy of GCS. In a pilot study eNO appeared to be superior to FEV1 in adjusting the dose of iGCS to control asthmatic children, but this needs to be confirmed with a larger sample size. Another non-invasive method to detect inflammatory markers is the technique of exhaled breath condensate (EBC). Although NO is degraded to NOx, it was found that eNO had no significant correlation with EBC NOx but had a significant correlation with pH. Hypertonic saline challenge, an artificial model of an asthmatic exacerbation was associated with an increase in EBC volume and the release of histamine, implicating mast cell activation. These novel findings suggest that non-invasive markers can be used both for clinical and mechanistic proposes.

Asthma is a chronic inflammatory condition of the airways characterised by reversible airflow obstruction, airway hyper-responsiveness and inflammatory infiltrates in the airway walls containing eosinophils, T lymphocytes and mast cells. T helper (Th) lymphocyte subsets, defined by the cytokines they secrete, are thought to play a key role in the in the initiation and perpetuation of chronic airway inflammation. Th2 cells, producing interleukin (IL)-4, IL-5, IL-9 and IL-13, are thought to be of particular importance. In contrast, Thl cells producing interferon (IFN)-y may counteract the development of Th2 responses and so down-regulate the asthmatic response. The prevalence of asthma is increasing but the reasons for this are not fully understood. In addition, some patients do not respond adequately to treatment with corticosteroids, currently the most effective anti-inflammatory agents used routinely in human asthma. There is therefore continual interest in developing new therapeutic agents for asthma. A greater understanding of the regulation of inflammatory responses in asthma will assist in the identification of potential targets for therapeutic intervention. The aims of this thesis were (i) to assess the role of the cytokine IL-18 in allergic airway inflammation by determining IL-18 levels in induced sputum in asthmatic subjects in comparison to normal subjects, and by studies in a murine model of allergic asthma using IL-18 gene deficient mice and (ii) to assess the potential antiinflammatory actions of simvastatin and thymosin beta 4 sulfoxide in the murine asthma model. IL-18 is a pro-inflammatory cytokine which can promote IFN-y secretion and, in association with IL-12, enhance the development of Thl responses. However, in some circumstances it may also stimulate Th2 responses. IL-18 therefore has the potential to suppress or exacerbate allergic airway inflammation. The role of IL-18 in both clinical and experimental asthma remains unclear. Statins are inhibitors of the rate-limiting enzyme, 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase, in cholesterol biosynthesis. As such they have been widely used as cholesterol lowering agents in clinical practice. They have previously been shown to have anti-inflammatory properties independent of their cholesterol-lowering ability in clinical studies of atherosclerotic disease and in animal models of Thlmediated inflammation. Thymosin beta 4 sulfoxide (T~4S0) is a 5 kDa peptide. Intracellularly its principal activity is to regulate actin polymerization. Corticosteroid treatment of monocytes in vitro induces the release of T~4S0 extracellularly, where it can inhibit neutrophil chemotaxis. Exogenous administration of T~4S0 has been shown to reduce neutrophilic inflammation in animal models. In this study it is shown that IL-18 is detectable in induced sputum fluid and IL-18 mRNA is expressed in induced sputum cells from asthmatic and nOlmal subjects. IL- 18 protein levels in induced sputum, and IL-18 mRNA expression in induced sputum cells were not significantly different between these groups. IL-18 production was localised to sputum macrophages. However, cigarette smoking significantly reduced IL-18 levels in induced sputum fluid in both asthmatic and normal subjects. In asthmatics, but not normal subjects, the reduction in IL-18 levels in sputum fluid was associated with reduced IL-18 mRNA expression in induced sputum cells. A murine model of allergic asthma, using BALB/C mice sensitised and challenged with ovalbumin (OVA), was used to examine the role of IL-18 in allergic responses in vivo. IL-18 gene knockout (ko) had significantly reduced bronchoalveolar lavage (BAL) total cell count and eosinophilia compared to wild-type (WT) mice. IL-18 ko mice had reduced IL-4 expression in thoracic lymph nodes, as assessed by quantitative peR, and significantly reduced OVA-specific IL-4 secretion from thoracic lymph node cultures assessed by ELISA. Serum OVA-specific IgG 1, IgG2a and IgE and total IgE levels were not significantly different between IL-18 ko and WT mice. The murine model of allergic asthma was also used to examine the anti-inflammatory activities of simvastatin and T~4S0 in a Th2-mediated, eosinophilic condition. Simvastatin treatment, either orally or intraperitoneally, and T~4S0 intraperitoneally reduced the total inflammatory cell infiltrate and eosinophilia in BAL fluid in response to inhaled OV A challenge. At higher doses of simvastatin intraperitoneally, a histological reduction in inflammatory infiltrates in the lungs was observed. Treatment with simvastatin intraperitoneally, but not orally, and T~4S0 were also associated with a reduction in IL-4 and IL-5 levels in BAL fluid. OVA-induced IL-4 and IL-5 secretion was reduced in thoracic lymph node cultures from both simvastatin-treated and T~4S0-treated mice. Neither simvastatin nor T~4S0 treatment altered serum total IgE or OVA-specific IgG 1 and IgG2a levels. The results described show that IL-18 can be detected in the induced sputum fluid of asthmatic and normal subjects and that cigarette smoking significantly reduces its levels. Studies in a murine model of allergic asthma suggest that IL-18 has a proinflammatory role in allergic airway inflammation, at least in part through its ability to induce IL-4 secretion. Both simvastatin and thymosin beta 4 sulfoxide had convincing anti-inflammatory properties in the murine model of asthma used, and these agents, or related compounds, may have therapeutic potential in human asthma.

[Truncated abstract] Asthma genetic studies have identified many genes that contribute to the pathogenesis of asthma and related variables. Members of the secretoglobin family appear to play an important role in controlling airway inflammation but they have received relatively little attention in asthma genetic research. In this thesis, I have investigated the genes of two members of the secretoglobin family (16 kDa Clara cell secretory protein (CC16) and secretoglobin 3A2 (SCGB3A2)) that are expressed at high levels in the airways and are important anti-inflammatory agents. The overall aim of these studies was to investigate the genetic variability of the CC16 and SCGB3A2 genes and their influence on airway inflammatory disease. The main hypothesis was that genetic variability in the genes for CC16 and SCGB3A2 exert an influence on airway inflammatory disease. Three populations were investigated: (1) a paediatric case control population (n=99), (2) an unselected birth cohort followed longitudinally at ages 1 month (n=244), six (n=123) and 11 years (n=195) and (3) an unselected Aboriginal Australian population (n=251). The case-control population was screened for novel DNA sequence variants in the CC16 promoter and the SCGB3A2 5’UTR and exons. No novel sequence variants were identified in the CC16 promoter and two were identified in the SCGB3A2 5’UTR (G- 811A and G-205A). A single nucleotide polymorphism previously identified in the CC16 gene (A38G) and the two polymorphisms identified in the SCGB3A2 gene were genotyped in both unselected populations. Genotype/phenotype associations were identified with adjustment for potential confounders such as age, gender, height and maternal tobacco smoking, where appropriate. This was due to the contribution of these factors to the aetiology of asthma, atopy and related phenotypes. All three polymorphism frequencies were significantly different between these two ethnically diverse populations

Asthma is a common and complex inflammatory disease of the airways characterized by deregulated immune responses that involves activation of multiple cell types including Th2 cells, IgE producing B cells, mast cells, basophils and eosinophils as well as resident lung cells such as epithelial, smooth muscle cells and macrophages. Despite intensive research, there are still unmet needs in the treatment of asthma. Recently, a new cytokine of IL 1 family, named IL 33 emerged as a potentially important factor in the immunopathogenesis of allergy and asthma. It was recently shown in our laboratory that intranasal administration of IL 33 can induce certain physiological features that are characteristic of experimental asthma, such as eosinophilic inflammation, Th2 cytokine and antibody production as well as increased airway hyperresponsiveness. The effect of IL 33 on the activation and differentiation of allergen specific Th2 cells has been well studied. However, the contribution of IL 33 to the activation of lung resident and inflammatory innate cells remains undefined. In this project I focused on alveolar macrophages and eosinophils as both cell types were reported to express IL 33R, ST2L and are thought to play a crucial role in asthma pathogenesis. I raised the hypothesis that IL 33 released locally in the lungs may trigger symptoms resembling asthma through the activation of airway alveolar macrophages. Furthermore, I hypothesize that IL 33 may exacerbate and maintain inflammation in the lungs by the direct activation of eosinophils. In our previous study we showed that IL 33 could switch the quiescent phenotype of alveolar macrophages toward the alternatively activated phenotype (M2, AAM). In the first part of my thesis I looked at the consequences of this phenomenon for airway inflammation. Using clodronate liposomes in vivo I was able to eliminate macrophage population from the lungs and demonstrated that resident alveolar macrophages are crucial for the development of IL 33 induced eosinophilic inflammation in the airways. I then examined the contribution of IL 13, a known M2 differentiation factor, to airway inflammation. Using anti IL 13 neutralizing antibodies I showed that IL 13 is required for the IL 33 triggered differentiation of alveolar macrophages toward M2 phenotype as well as for eosinophilic inflammation. Next, I looked at how IL 33/ST2 pathway modulates the differentiation and activation of eosinophil. I demonstrated that bone marrow hematopoietic progenitors CD117+ express ST2L and that IL 33 is able to differentiate these cells toward eosinophils. By employing deficient mice or neutralizing antibodies I found that this process is ST2 and IL 5 dependent and independent of IL 13. I then extended my research interests to include mature mouse and human eosinophils. I showed that both human and mouse resting eosinophils express low levels of ST2L which can be markedly increased by IL 33. Moreover, I demonstrated that eosinophils that are recruited to the lungs during experimental allergic airway inflammation express high levels of ST2L. Furthermore, I carried out a study on effector function of eosinophils. I found that IL 33 induces IL 13, IL 6 and increases TARC, TGF production by mouse eosinophils. In addition, IL 33 exacerbated IgG induced human and mouse eosinophil degranulation, likely by enhancing FcRII expression. Having shown earlier that IL 13 is requited for the polarization of alveolar macrophages toward AAM by IL 33 in vitro and in light of the fact that IL 33 stimulated eosinophils can be a significant source of IL 13; I went on to investigate the interaction between macrophages and eosinophils. Using co cultures of ST2 / macrophages with WT eosinophils in Transwell system, I demonstrated that IL 33 but not IL 5 activated eosinophils can support macrophage polarization toward the pro inflammatory AAM phenotype, partially through the production of IL 13. Finally, given the role of IL 33/ST2L axis in eosinophil activation in vitro, I investigated the contribution of IL 33 activated eosinophils to airway inflammation in vivo. Using adoptive transfer protocol I showed that the contribution of IL 33 activated eosinophils to airway inflammation is mediated primarily by the release of cytokines from these cells which, in turn, recruits other inflammatory cells and supports the differentiation of alveolar macrophages towards AAM. These data show that IL 33/ST2 pathway regulates multiple features of alveolar macrophage and eosinophil biology that can have a significant impact on asthma pathophysiology in the airways. Studies carried out in our laboratory and elsewhere suggest that IL 33 is equally capable of activating other cell types that have been implicated in asthma pathology such as Th2, B1 cells, DCs, mast cells and basophils. Therefore, targeting IL 33/ST2 pathway may potentially offer a promising therapeutic approach to asthma and allergy.

Although the aetiology of chronic cough in guidelines is clearly stated as asthma and related syndromes, gastro-oesophageal reflux disease, and upper airways disease, the inflammatory mechanisms underlying these conditions differ. Recent studies on asthma have increasingly focused on its molecular phenotypes instead of clinical characteristics. Predominantly in this thesis I hypothesize that by dividing cough patients into the clinical characteristics of eosinophilic and neutrophilic groups will enhance our ability to recognise the type of airway inflammation, and consequently will lead us to more targeted treatment approaches. To investigate this hypothesis I conducted a randomized, single centre, open label, controlled, clinical trial to examine the outcome of anti-inflammatory therapy with either montelukast or prednisolone in 50 patients with chronic cough. Furthermore, I studied the epidemiology of 137 chronic cough patients attending the Hull cough clinic. Results from the clinical study demonstrated that patients with FeNO≤20ppb had twice the number of coughs compared with patients with FeNO≥30ppb. This was reflected on quality of life as assessed by the LCQ and HARQ. Confirming this finding I found in the epidemiological study, that patients attending the hull cough clinic with FeNO≤25ppb scored significantly higher in HARQ compared with FeNO≥25ppb. In the clinical trial study I have shown that FeNO was a good marker for eosinophilic inflammation. There was a high degree of correlation with FeNO, blood and sputum eosinophilia thus confirming phenotypic identity. Whether the FeNO can be used to identify the different characteristics between eosinophilic and non-eosinophilic coughs needs further investigation. Cough patients in both low and high FeNO groups have shown a similar response to montelukast despite anticipating little or no effect in those without eosinophilic inflammation. These results suggest that response to montelukast may not be predicted by presence of eosinophilic biomarkers alone but may be act by effecting localised leukotriene mediated inflammation.

Air pollutants, such as ozone (O3) and diesel exhaust particles, elicit oxidative stress in the lung. Antioxidants within the respiratory tract lining fluid (RTLF) protect the underlying tissue from oxidative injury. Supplementation with vitamins has been shown to modulate the acute ozone-induced effects, but the mechanisms behind this have not been fully clarified. The aim of this thesis was to investigate the airway responses to diesel exhaust and ozone exposure in healthy humans, with the emphasis on inflammatory and antioxidant responses. Furthermore, to study whether oral supplementation with vitamin C could increase ascorbate concentration in the RTLF and whether vitamin supplementation could modulate the negative effects induced by ozone exposure. Diesel exhaust (100 µg/m3 PM10 for 2h), evaluated 18 hours post exposure (PE), induced a neutrophilic airway inflammation and an increase in bronchoalveolar (BAL) urate and reduced glutathione. During O3 exposure (0.2 ppm for 2h), significant losses of nasal RTLF urate and ascorbate concentrations were observed. Six hours PE, a neutrophilic inflammation was evident in the bronchial wash (BW), together with enhanced concentrations of urate and total glutathione. In the bronchoalveolar lavage (BAL), where vitamin C, urate and glutathione concentrations were augmented, no inflammatory response was seen. In alveolar lavage leukocytes, there was a significant loss of glutathione and cysteine, whereas an increase in ascorbate was found in bronchial tissue samples. Following supplementation with increasing doses of vitamin C (60-1,000 mg/day, for 14 days), evaluated 24 hours after the last dose, ascorbate concentrations were unchanged in the nasal RTLF, despite elevated concentrations in plasma and urine. In contrast, following a single dose of 1g of vitamin C, vitamin C concentrations increased significantly in both plasma and nasal lavage two hours post supplementation, before returning to baseline levels at 24 hours. Notably, dehydroascorbate (DHA) accounted for the largest part of RTLF vitamin C and a number of control experiments were performed to ensure the authenticity of this finding. Healthy O3 responders were exposed to O3 (0.2 ppm for 2 h) and air, following seven days of supplementation with vitamin C and E or placebo. No protective effect on lung function or airway inflammation was observed following supplementation. BW and BAL-DHA were enhanced after O3, with further increases following supplementation. In conclusion, oxidative air pollutants induce airway inflammation, as well as a broad spectrum of antioxidant adaptations, which could ultimately limit the airway inflammatory responses. Oral vitamin supplementation was shown to augment RTLF-vitamin C concentrations, but it did not provide protection from the ozone-induced airway responses following a single insult of ozone. The finding of high concentrations of DHA in the RTLF could indicate that DHA represents an important transport form of vitamin C onto the surface of the lung.

Smokers with asthma represent an important sub-group of asthmatics displaying both reduced response to inhaled and oral corticosteroids as well as demonstrating accelerated decline in lung function and increased use of health care services. Clinical and laboratory studies have suggested that macrolide antibiotics may exhibit anti-inflammatory properties in a variety of airways disease including asthma. The anti-inflammatory properties of macrolides have been recognised for almost 50 years. Indirect evidence from both pre-clinical and clinical studies suggests that the mechanism of action may be of particular benefit in smokers with asthma. A proof of concept study was designed to test the hypothesis that the macrolide antibiotic azithromycin improves measures of asthma control, airway inflammation and bacterial colonisation in smokers with asthma. Azithromycin was chosen for its convenience of once daily dosing and its oral tolerability in addition to its more limited interactions. Seventy-seven adults with allergic asthma were recruited to a 12-week parallel group randomised controlled trial comparing the effects on asthma control, airway inflammation and bacterial colonisation of oral azithromycin 250 mg daily with matched placebo. The primary outcome measure was peak expiratory flow at the final study visit. Secondary outcome measures included spirometry, asthma control questionnaire [ACQ] score, asthma quality of life questionnaire [AQLQ], Leicester cough questionnaire [LCQ] score, provocation concentration to methacholine PC20, and inflammatory markers: exhaled nitric oxide, sputum differential cell counts, sputum supernatant and serum inflammatory markers such as interleukin-1β [IL-1β], IL-2, -4, -5, -6, -10, TNF-α, IFN-γ, GM-CSF, Leukotriene B4, and high sensitivity C-reactive protein. Microbiological culture and PCR of sputum was also performed to assess for any changes associated with treatment. At 12 weeks, the change in PEF at the final study visit, as compared with baseline, did not differ significantly between the azithromycin and placebo treatment groups [mean difference azithromycin-placebo -10.3L/min, 95% CI -47.1 to 26.4, p=0.58]. No statistically significant difference was observed between the azithromycin and placebo groups in each of the measures of spirometry, ACQ, AQLQ, LCQ, PC20, or evening PEF. The LCQ-psychological domain did reach statistical significance, [mean difference azithromycin-placebo -0.46, 95%CI -0.9 to 0.02 p=0.04], however this indicates a deterioration in the treatment group. No change was seen in exhaled nitric oxide. The total cell counts recovered from sputum were similar following treatment with azithromycin compared to placebo. In addition, differential cell counts remained unchanged and lymphocyte proliferation assays did not demonstrate any statistically significant changes following 12 weeks of treatment with azithromycin when compared to placebo. There was no substantial difference in any of the measured sputum supernatant or plasma cytokines. Peripheral blood monocyte stimulation was performed, with supernatant being measured against a panel of cytokines. There was again no substantial difference in any of the measured panel of cytokines collected from the monocyte stimulation assays when the azithromycin group was compared to placebo. There was no correlation between changes in ACQ, AQLQ, LCQ, PC20, sputum macrophage count, sputum neutrophil count, sputum eosinophil count, and PEF. Adverse event rates were similar in patients taking azithromycin compared with placebo. A total of 4 patients were lost to follow up [1 in the azithromycin group, 3 in the placebo group]. One patient died of a cardiovascular cause. This occurred following completion of the study but within the pre-specified regulatory reporting period. In conclusion there were no clinically important improvements in a range of clinical indices of asthma control, airway inflammation or bacterial colonisation following 12 weeks treatment with azithromycin when compared with placebo in smokers with asthma. The lack of any evidence of clinical benefit of azithromycin in smokers with asthma is a new finding and extends the current knowledge base and evidence for the use of macrolides in asthma. There exists no firm evidence to suggest the widespread use of macrolides in asthma and the current study suggests that no benefit will be observed in the sub-group of asthmatics whom are current smokers.

Obliterative bronchiolitis (OB) is the leading cause of chronic irreversible lung graft failure. It is assumed by many to be caused by chronic rejection. We hypothesized that non-specific airway inflammation induced by factors such as foreign airway deposited particles, antigens and microbes is essential for the development of OB after lung transplantation (LTx). In this study, we examined the histological and immunohistological changes in allografted airways following transtracheal installation of foreign irritant particles.
Inadequate immunosuppression of lung allografts leads to severe vascular changes consistent with chronic vascular rejection but fails to induce airway inflammation and OB. The addition of a nonspecific foreign irritant was necessary to induce diffuse airway inflammation and OB in our animal model. This suggests that a co-factor inducing airway inflammation, such as deposited foreign particles or infections, is likely necessary for the development of OB after lung transplantation. Cytokine expression reveals a link between the observed influx of airway inflammatory cells and IL-8. While bFGF is associated more with the proliferative phases of airway and vascular injury. These markers however are not predictive of either rejection or OB, due to the expression of both these markers in all three study groups. (Abstract shortened by UMI.)

Tissue eosinophilia is a prominent feature of allergic inflammatory diseases that may be in part mediated through the regulation of eosinophil survival at inflammatory site. T helper 2 type cytokines, such as interleukin (IL)-5, Granulocyte Macrophage Colony Stimulating Factor (GM-CSF), are the important mediators of allergic inflammation leading to prolonged survival of eosinophils. Due to the proposed central role of eosinophils in asthma and other allergic airway diseases, there is considerable interest, to determine the mechanisms that regulate eosinophil functions and accumulation. IL-12 and IL-18 attract considerable attention for their immunomodulatory roles on T helper cell subsets, and their potential to affect on eosinophil function.
The general aim of this study was to determine the effect of IL-12 and IL-18 on eosinophil functions. Results from this thesis demonstrate that eosinophils express functional receptors for IL-12 and IL-18. Acting alone or in synergy, IL-12 and IL-18 induced eosinophil apoptosis, in vitro. The apoptotic effect of IL-12 was reversed by IL-5, suggesting that IL-5 and IL-12 have counter-regulatory effects on eosinophil survival. Our regulation studies demonstrated that Phorbol-Myristate-Acetate (PMA) induced optimal expression of IL-18 and IL-12 receptors by eosinophils. IL-18 receptor expression by eosinophils was markedly increased following stimulation with interferon (IFN)-gamma, Tumor Necrosis Factor (TNF), or IL-12. Up-regulation of IL-18 receptor upon IL-12 stimulation was particularly important, which may explain IL-12 and IL-18 synergy on eosinophil apoptosis. We also investigated IL-12 and IL-18 expression by eosinophils. Eosinophils did not express IL-18. However, there was constitutive IL-12 expression by eosinophils. Release of IL-12 was also confirmed in eosinophil supernatants, which suggested an autocrine mechanism of IL-12 action on eosinophils.
Extending our understanding of the role of IL-12 and IL-18 in allergic inflammation, we have defined a novel pathway by which IL-12 and/or IL-18 may actually regulate eosinophils functions, and exert an inhibitory effect on eosinophil survival. Our findings provide critical new insights into mechanisms regulating eosinophil survival. To gain an even more detailed understanding of regulatory signals mediating eosinophil survival, we need to define the mechanisms involved in IL-12 and IL-18 signalling. We have just begun to identify the molecules involved, and these include IL-12 and IL-18 receptors. Activation via IL-12 and/or IL-18 receptor-mediated mechanisms may provide a novel strategy for reducing the numbers or inhibiting the function of these cells in allergic diseases or other diseases characterized by increased numbers of, or mediators from, eosinophils.

Background: Ambient air pollution is associated with adverse health effects, but the sources and components, which cause these effects is still incompletely understood. The aim of this thesis was to investigate the pulmonary effects of a variety of common air pollutants, including diesel exhaust, biomass smoke, and road tunnel and subway station environments. Healthy non-smoking volunteers were exposed in random order to the specific air pollutants and air/control, during intermittent exercise, followed by bronchoscopy. Methods and results: In study I, exposures were performed with diesel exhaust (DE) generated at transient engine load and air for 1 hour with bronchoscopy at 6 hours post-exposure. Immunohistochemical analyses of bronchial mucosal biopsies showed that DE exposure significantly increased the endothelial adhesion molecule expression of p-selectin and VCAM-1, together with increased bronchoalveolar lavage (BAL) eosinophils. In study II, the subjects were exposed for 1 hour to DE generated during idling with bronchoscopy at 6 hours. The bronchial mucosal biopsies showed significant increases in neutrophils, mast cells and lymphocytes together with bronchial wash neutrophils. Additionally, DE exposure significantly increased the nuclear translocation of the aryl hydrocarbon receptor (AhR) and phosphorylated c-jun in the bronchial epithelium. In contrast, the phase II enzyme NAD(P)H-quinone oxidoreductase 1 (NQO1) decreased after DE. In study III, the 2-hour exposures took place in a road tunnel with bronchoscopy 14 hours later. The road tunnel exposure significantly increased the total numbers of lymphocytes and alveolar macrophages in BAL, whereas NK cell and CD56+/T cell numbers significantly decreased. Additionally, the nuclear expression of phosphorylated c-jun in the bronchial epithelium was significantly increased after road tunnel exposure. In study IV, the subjects were exposed to metal-rich particulate aerosol for 2 hours at a subway station with bronchial biopsy and BAL sampling at 14 hours. The subway exposure significantly increased the concentration of glutathione disulphide (GSSG) in BAL, with no airway inflammatory responses. In contrast, the number of neutrophils in the bronchial mucosa and the nuclear expression of phosphorylated c-jun in the bronchial epithelium tended to decrease after the subway exposure. In study V, the exposure to biomass smoke lasted 3 hours. Bronchoscopy was conducted 24 hours post exposure. The investigated biomass combustion emissions resulted in a significant increase in total glutathione and reduced glutathione in BAL, without any evident acute airway inflammatory responses.     Conclusion: The present thesis presents data from exposures of healthy subjects to a variety of common air pollutants, as compared with an air reference. Oxidative as well as bronchial mucosal and bronchoalveolar responses differed between these air pollutants, with the most pronounced airway effects seen after exposure to diesel exhaust. This may be due to differences in pulmonary deposition, physicochemical characteristics, toxicological pathways and potency. Additional studies will assist in addressing dose-response and time kinetic aspects of the airway responses.

[Tuncated abstract] There has been longstanding concern over whether endobronchial biopsies adequately represent inflammation throughout the bronchial tree in diseases such as asthma, despite the endobronchial biopsy technique having been used frequently to assess airway inflammation in research settings. There has also been ongoing debate about whether endobronchial biopsies should be assessed by new, unbiased, three-dimensional (3D) stereological techniques instead of traditional, two-dimensional (2D) non-stereological techniques. Therefore, the aims of this study were: (i) to investigate whether endobronchial biopsies represent the density of mast cells in the large and small airways, in alveolar walls and in the lung as a whole (ii) to use both stereological and non-stereological methods to address this question, and where possible, to compare the results of these two approaches. '...' Mast cell density in biopsies was not related to mast cell density immediately adjacent to the biopsy site or to mast cell density in the total airway wall in the large airways, the inner airway wall in the small airways, the walls of the alveoli or the lung as a whole. In general, measurements of mean mast cell density on biopsies to a depth of 100µm below the basement membrane were poorly related to mean mast cell density in other compartments of the lung. Mean 3D and 2D mast cell densities were strongly correlated (r 0.9, p < 0.005) and where both methods were used, results were similar. The mean height and area profile of a mast cell were approximately 12µm and 68µm2 respectively. In disk-shaped IUR lung samples, percent shrinkage in height due to paraffin processing was systematically greater than percent radial shrinkage by an average of approximately 4 times. Cavalieri lung volumes were systematically smaller than displacement volumes by an average of 14%. Any given endobronchial biopsy is unlikely to represent mast cell density around the airway wall generally in the vicinity of the biopsy site. However, the average of at least 4 biopsies from different sites in the proximal airways can be used to both represent mean mast cell density in the inner airway wall of the large airways, and act as the basis for inter-subject comparisons of mean mast cell density in the total airway wall of the small airways. On biopsies, mast cell counts should be measured over the entire inner airway wall not just to a depth of 100µm or less below the basement membrane. 3D mast cell densities obtained by stereological methods are closely related to 2D mast cell densities obtained by non-stereological methods and are likely to result in similar conclusions. Lung volumes are smaller when measured by the Cavalieri method than when measured by fluid displacement. Shrinkage of isotropic uniform random samples of human lung tissue due to paraffin processing is anisotropic. The mean volume of a mast cell in the human lung is likely to be much smaller than that reported previously for monkey lungs.

Thesis (Ph.D.)--Indiana University, Dept. of Kinesiology, 2009.
Title from PDF t.p. (viewed on Jul 19, 2010). Source: Dissertation Abstracts International, Volume: 70-12, Section: B, page: 7486. Adviser: Timothy D. Mickleborough.

Asthma is a chronic respiratory disease characterized by inflammation of the airway and the presence of recurrent attacks (exacerbations) of breathlessness, wheezing, cough, chest tightness, or some combination of these symptoms. In the US, about 53% of people with asthma had an asthma attack in 2008, and 57% of these, were children. One in ten children (10%) had asthma in 2009, and boys were more likely than girls to have asthma. Internationally, the prevalence of asthma varies widely in different countries, but the disparity is narrowing due to rising prevalence in low and middle income countries. Unfortunately, we do not have statistics for asthma in the Republic of Panama, neither epidemiological data nor costs, which is the reason why this research is needed. The Panamanian Social Security Fund (CSS) provides protection to workers, their immediate families and the pensioned. By the end of 2010, the total insured population was 2,862,202 (83% of the total population of Panama). Of the total insured population 58% were dependent. Of this, 1,205,607 (42%) were children. On the basis of this information, we decided to develop the research study using information from the CSS, specifically in the Hospital de Especialidades Pediatricas (HEPOTH). It is the only tertiary level of healthcare children's hospital of the CSS. A quantitative-descriptive design was used to develop this study. Data was collected from medical records of patients diagnosed with asthma in the HEPOTH from January to June 2012. We reviewed the medical records of each care area by month, and numbered each clinical record of children diagnosed with asthma in crisis and randomly selected 10% of the medical records from a minimum of 2000 records. Information on treatment costs was also obtained. Once all the information was collected, it was typed in the digital data log created for this study and the responses were code converted and the information was entered into a database. The data were exported to IBM SPSS Statistics 21. The average cost of asthma attacks in Panama is estimated at $205.52. We were able to confirm that there are variations in this average by gender, age, geographic area of residence, season, severity, whether treated in the emergency department or hospitalization, and the type of treatment received. It was also possible to obtain secondary information about the epidemiology of asthma that allowed us to confirm that our statistics matched international statistics.

Astma yra lėtinė kvėpavimo takų uždegimo liga. Mokslininkai neabejoja, jog 2-o tipo T limfocitai pagalbininkai bei eozinofilinis kvėpavimo takų uždegimas yra astmos patogenezės pagrindas. Tačiau šis mechanizmas ne visuomet gali paaiškinti astmos metu esančio kvėpavimo takų uždegimo bei klinikinių simptomų įvairumą, eigos ypatumus ir net skirtingą atsaką į gydymą. Eksperimentiniai tyrimai su gyvūnais parodė 17-o tipo T limfocitų pagalbininkų (Th17) svarbą alerginės astmos vystymuisi. Todėl šio mokslinio tyrimo tikslas buvo įvertinti Th17 limfocitų vaidmenį sergant alergine astma. Tyrimo metu sergantiesiems alergine astma nustatytas didesnis Th17 limfocitų kiekis periferiniame kraujyje bei didesnė interleukino (IL)-17 koncentracija serume ir indukuotuose skrepliuose, lyginant su sveikais asmenimis. Be to, bronchų provokacija su Dermatophagoides pteronyssinus alergenu sukėlė Th17 limfocitų ir IL-17 kiekio padidėjimą praėjus 7 ir 24 val.po jos, ypatingai ryškų sergantiems alergine astma su ankstyva ir vėlyva bronchų obstrukcija. Atlikto tyrimo rezultatai parodė, kad Dermatophagoides pteronyssinus alergenas sukelia vietinį ir sisteminį Th17 limfocitų imuninį atsaką kuris yra susiję su vėlyvos fazės kvėpavimo takų uždegimu.
Allergic asthma is a chronic inflammatory disease of the airways associated with the response of predominant type 2 helper T lymphocytes to an inhaled allergen. However, differences in inflammation and clinical symptoms of this disease not always can be explained by this mechanism. Recent animal model studies have shown the importance of type 17 helper T lymphocytes (Th17) in the development of allergic asthma. The role of these cells in causing allergen-induced airway inflammation as well as systemic inflammatory response in human is still not well defined. Therefore, we investigated the peripheral blood Th17 lymphocyte response to inhaled Dermatophagoides pteronyssinus (D. pteronyssinu) allergen in patients with allergic asthma. The present study has shown that patients with allergic asthma have a higher percentage of peripheral blood Th17 lymphocytes and elevated serum as well as induced sputum interleukin-17 levels compared with healthy subjects. Moreover, all studied allergic asthma patients, especially with early- and late-phase asthmatic reaction, showed an enhanced airway and systemic Th17 lymphocyte response 7 h and 24 h after bronchial challenge. We documented an enhanced local and systemic Th17 lymphocyte response to inhaled D. pteronyssinus in association with late-phase allergen-induced airway inflammation in patients with allergic asthma.

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FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais
A asma alérgica é uma doença inflamatória crônica das vias aéreas, caracterizada por uma resposta de hipersensibilidade imediata, obstrução brônquica, inflamação pulmonar e níveis elevados de IgE. A doença é mediada principalmente por uma resposta imunológica alérgeno-específica tipo Th2. Nas últimas décadas, a prevalência da asma alérgica tem aumentado significativamente, sobretudo nos países desenvolvidos. A Hipótese da Higiene atribui este aumento a uma menor exposição a determinados microrganismos durante a infância, quando o amadurecimento adequado do sistema imunológico requer estímulos que induzam respostas imunológicas de perfil Th1, fundamentais para o equilíbrio de respostas Th2 exacerbadas. Diversos trabalhos epidemiológicos parecem comprovar esta hipótese, evidenciando a existência de uma relação inversa entre o contato com microrganismos indutores de uma resposta Th1 e o desenvolvimento de asma alérgica. Paralelamente, estudos em modelos murinos constataram que o tratamento com Mycobacterium bovis BCG (BCG) reduz respostas Th2 alérgenoespecíficas. No entanto, os mecanismos pelos quais a micobactéria inibe o desenvolvimento da resposta alérgica são ainda pouco conhecidos. Este estudo avaliou o efeito da administração do BCG sobre a resposta imunológica ocorrida na alergia pulmonar em camundongos BALB/c previamente sensibilizados e desafiados com OVA. Vinte e quatro horas após o último desafio, o sangue e o lavado broncoalveolar foram coletados para análises de imunoglobulinas e contagem de células, respectivamente. Adicionalmente, os pulmões foram submetidos à análise histológica, avaliação da atividade de EPO e dosagens de citocinas e quimiocinas, assim como avaliação da expressão de CTLA-4, Foxp3 e IL-10 por citometria de fluxo. Os resultados obtidos indicam que o tratamento com BCG melhorou o processo alérgico através da redução dos principais parâmetros relacionados à resposta Th2, como o infiltrado eosinofílico pulmonar, a atividade de EPO, IL-4, IL-13, CCL11, além de IgE e IgG1 específicas anti-OVA. Por outro lado, a administração da micobactéria aumentou os níveis de IFN-γ, IL-10 e TGF-β, além das expressões de Foxp3 e CTLA-4 pelos linfócitos T CD4+. Paralelamente, houve um aumento na produção de IL-10 pelos linfócitos T CD8+. Esses dados sugerem que, além da indução de uma resposta imune Th1, a ação imunomoduladora do BCG está relacionada também à indução de mecanismos reguladores.
Atopic asthma is a chronic respiratory disease characterized by airway hyperresponsiveness, reversible airway obstruction, lung inflammation, and high levels of allergen-specific IgE, driven by allergen-specific Th2 cells. The increasing prevalence of allergic diseases, particularly in industrialized countries, has led to the hygiene hypothesis, which states that the newborn infant’s immune system is skewed toward Th2 responses and needs timely and appropriate environmental stimulus to create a balanced immune response. Supporting this hypothesis, epidemiological and experimental evidence has shown an inverse correlation between Th1-induced microbial infections and atopic asthma. Similarly, some animal studies have demonstrated that exposure to Mycobacterium tuberculosis or to environmental mycobacteria is able to protect against the development of allergic responses. However the exact mechanism underlying this inhibition still remains poorly understood. This study aimed to evaluate the ability of BCG to suppress an established allergic response in a mouse model of OVA-induced airway inflammation. To achieve this, OVA sensitized and challenged BALB/c mice were twice treated with BCG via nasal and 21 days after the first treatment, mice were rechallenged with OVA. Twenty-four hours after the last challenge, blood samples were collected to detect anti-OVA immunoglobulin isotypes, and bronchoalveolar lavage (BAL) was harvested for cell count. Additionally, lungs were collected for histological analysis, detection of EPO activity and measurement of cytokines and chemokines. The expression of CTLA-4, Foxp3 and IL-10 was also determined in lung tissue by flow cytometry. The data indicated that BCG treatment was able to inhibit an established allergic Th2-response by decreasing the allergen-induced eosinophilic inflammation, EPO activity, levels of IL-4, IL-13, CCL11 and serum levels of IgE and IgG1. Mycobacteria treatment increased lung levels of IFN-γ, IL-10 and TGF-β, and expressions of Foxp3 and CTLA-4 in CD4+T cells. Additionally, an increased production of IL-10 by CD8+ T cells was observed, even though no detectable changes in CD4+IL-10+ was noticed. Altogether, these results suggest that the mechanism underlying the down-regulatory effects of BCG on OVA-induced airway inflammation appear to be associated with the induction of both Th1 and T regulatory immune responses.

Background Nitric oxide (NO) was well known to be a component of air pollution, often in the form of nitrogen dioxide (NO2). However its importance in biological systems altered dramatically with the discovery in 1987 that it was the 'endothelial-derived relaxing factor'. Since then there has been an explosion of research on NO demonstrating that this gaseous molecule was a widespread physiological mediator and was simultaneously recognised as a vital component of immune function contributing to macrophage-mediated cytotoxicity. NO was therefore a key molecule in modulating inflammation, including airway inflammation. The aim of this thesis was: 1. To adapt a NO chemiluminscence analyser from measuring airway pollution to measuring exhaled air in human subjects. 2. To measure NO levels in exhaled air in adult subjects. 3. To evaluate whether altering measurement parameters altered the levels of NO obtained. 4. To adapt this technique from adults to measure exhaled NO in children. 5. To compare levels of NO from healthy children to groups of asthmatic children on either bronchodilator therapy only, or on regular inhaled corticosteroids. 6. To compare the levels of NO in a pilot group of asthmatic children before and after commencement of inhaled corticosteroids. Methods A Dasibi Environmental Corporation Model 2107 chemiluminescence analyser was adapted specifically requiring a reduction in response time, which was achieved by modification of the circuitry and re-routing of the analogue signal directly to a chart recorder, achieving a reduction of the response time by 80%. Addition of a number of analysers allowed the measurement of exhaled NO, carbon dioxide (CO2), mouth pressure and flow for each exhalation from total lung capacity. Twenty adult subjects (in total) were then studied looking at direct (NO, CO2, mouth pressure) versus t-piece (with the addition of flow) measurements making five exhalations from total lung capacity, at 3-minute intervals (direct/t-piece/direct or t-piece/direct/t-piece in series). The area of NO under the curve versus the peak of the NO trace was compared and the exhalation pattern of NO versus CO2 was compared. Measurement conditions were altered to evaluate the effect of individual parameters on the exhaled NO result. This included separately assessing different expiratory flows, different expiratory mouth pressures, the effect of a high versus a low background NO level and the effect of drinking water (of varying temperatures) prior to exhalation. Healthy control children were then enrolled to the study from a local school (Park Walk Primary School) and compared with asthmatic children enrolled from outpatient clinics at the Royal Brompton Hospital. The asthmatic children were further divided into those on bronchodilator treatment only and those on regular inhaled corticosteroid therapy. NO was also measured before and two weeks after commencing inhaled corticosteroid therapy in previously steroid-naive asthmatics. Results It was possible to modify a chemiluminescence analyser to enable measurement of exhaled NO. In 12 healthy subjects (mean age 32 years, 6 males) peak direct NO levels were 84.8 parts per billion (ppb) (standard error of the mean (SEM) 14.0ppb), significantly higher than 41.2ppb (SEM 10.8ppb) measured via the t-piece system. The exhaled NO rose to an early peak and plateau while the CO2 levels continued to rise to peak late in the exhalation. The mean times to peak NO levels were 32.2 seconds (s) (direct) and 23.1s (t-piece), which was significantly different from the mean times to peak CO2 levels at 50.5s (direct) and 51.4s (t-piece, p<0.001). At peak NO level, the simultaneous CO2 level of 4.9% (SEM 0.47%, direct) and 5.2% (SEM 0.18, t-piece) were significantly lower than the peak CO2 achieved of 5.8% (SEM 0.21%, direct, p<0.001) and, 6.2% (SEM 0.28, t-piece, p<0.001). There was no difference between repeat direct or t-piece measurements. With regard to varying measurement conditions, the mean peak concentrations of NO decreased by 35ppb (95% confidence intervals 25.7-43.4) from a mean of 79ppb (SEM 15.4ppb) at an expiratory flow rate of 250mls/min to 54.1ppb (SEM 10.7ppb) at 1100mls/min. The mean peak concentration of NO did not change significantly when mouth pressure was increased in eight of ten subjects, although in two it did decrease in the highest pressure. The mean NO concentration with machine and subjects sampling from a low NO reservoir was 123ppb (SEM 19.4), which was an increase from results obtained before at 81.9ppb, SEM 10.2ppb, p=0.001 95%, CI -19.9 to -62.7) and after at 94.2ppb(SEM 18.3ppb, p=0.017, 95% CI 6.0-5.18) sampling with high ambient NO levels. The mean peak NO concentration decreased from 94.4ppb (SEM 20.8) to 70.8ppb (SEM 16.5, p=0.002 95% CI 12.9 -33.1) with water consumption. In 39 healthy pre-pubertal children with a mean age of 9.9 years (range 9-11 years, 23 girls) the mean direct exhaled No level was 49.6ppb (SD 37.8ppb, range 11.5-197.2ppb) compared with mean exhaled No via t-piece sampling of 29.2ppb (SD 27.1ppb, range 5.1-141.2ppb). There was no significant difference between boys and girls for either the direct or the t-piece recordings. In comparison with normal children, 15 asthmatic children on bronchodilator therapy only had much higher levels of exhaled NO at 126.1ppb via the direct system (SD 77.1ppb, p<0.001) and 109.5ppb via the t-piece system (SD 106.8ppb, p<0.001). In 16 asthmatics on regular inhaled corticosteroids the mean peak exhaled levels were significantly lower at 48.7ppb via the direct method (SD 43.3ppb, p<0.001) and via the t-piece system at 45.2ppb (SD 45.9ppb, p<0.01). There was no difference between the normal children and the asthmatic children who were on regular inhaled corticosteroids (p=0.9 direct, p=0.2 t-piece).There was no significant difference in CO2, mouth pressure, duration of expiration and expiratory flows between the three groups or between the two methods (direct and t-piece). In six asthmatic children the mean peak exhaled NO levels fell from a medium peak level of 124.5ppb to 48.6ppb when measured before and two weeks after commencement of inhaled corticosteroids on treatment. Discussion This research showed it was possible to modify an NO chemiluminescence analyser to enable measurement of exhaled NO in adult and paediatric subjects. Furthermore, it was possible to measure both healthy and asthmatic children. There were significant differences between the exhalation pattern of NO and CO2 suggesting that NO was produced in the airways, not at alveolar level, unlike CO2. The measurement of exhaled NO required a standardised approach as exhaled NO levels decreased with increasing expiratory flow, when measuring at a time of high ambient NO concentration, and with consumption of either hot or cold water immediately preceding exhalation (such as might be given if a subject was coughing). The findings with expiratory mouth pressure were less certain, with a difference seen in only two of ten subjects. The levels of exhaled NO measured in children aged 9-11 years were lower than that measured in the adult subjects. There was no difference between boys and girls, or with other parameters such as having a personal history of atopy, a family history of atopy, or the presence of a smoker or furry pets within the house-hold. These findings may have altered with increased numbers in this group and could possibly be a type two statistical error. The results of exhaled NO in asthmatic children on bronchodilator therapy only were significantly elevated compared to both normal children and asthmatic children treated with regular inhaled corticosteroids. The exhaled NO level also fell significantly by two weeks following the commencement of inhaled corticosteroid treatment in steroid-naive asthmatic children. These results suggested that the methods of measuring exhaled NO required standardization and that it could potentially be a non-invasive measure of airway inflammation to follow - particularly in children with asthma who were commencing inhaled steroid treatment.

The Streptococcus pneumoniae-derived toxin, pneumolysin, has been reported to augment neutrophil-mediated inflammatory responses in murine models of experimental infection of the airways, and to favour invasive pneumococcal disease. The laboratory research presented in this thesis has been designed to investigate the possible proinflammatory interactions of pneumolysin with human neutrophils in vitro, as well as the underlying mechanisms of these. Addition of pneumolysin (0.0167 - 41.75 ng/ml) to neutrophils caused dose-related enhancement of the following proinflammatory activities of these cells: superoxide generation, elastase release, expression of the β2-integrin CR3, phospholipase A2 activity and production of leukotriene B4 and prostaglandin E2, oxidative inactivation of α-1-proteinase inhibitor, and synthesis and release of interleukin-8. Pneumolysin-mediated enhancement of these neutrophil activities was observed in the absence of detectable cytotoxicity and was most striking when the toxin was added together with the bacterial chemoattractant N-formyI-L-methionyl-L-leucyl-L-pnenylalanine (FMLP, 1 µM). Treatment of neutrophils with pneumolysin also resulted in uncontrolled influx of Ca2+ into the cells in the setting of membrane depolarisation and efflux of K+, which appeared to be a consequence of the pore forming actions of the toxin. Importantly, the proinflammatory interactions of pneumolysin with neutrophils were completely attenuated by exclusion of Ca2+ from the cell-suspending medium. These observations identify novel proinflammatory properties of pneumolysin which result from pore formation in the plasma membrane, influx of Ca2+ and augmentation of Ca2+ -activitable neutrophil functions.
Thesis (DPhil)--University of Pretoria, 2005.
Immunology
unrestricted

Indiana University-Purdue University Indianapolis (IUPUI)
More than 300 million people world-wide suffer from breathlessness, wheezing, chest tightness, and coughing characteristic of chronic bronchial asthma, the global incidence of which is on the rise. Allergen-sensitization and challenge elicits pulmonary expression of chemoattractants that promote a chronic eosinophil-rich infiltrate. Eosinophils are increasingly recognized as important myeloid effectors in chronic inflammation characteristic of asthma, although few eosinophil molecular signaling pathways have successfully been targeted in asthma therapy. p21 activated kinases (PAKs), members of the Ste-20 family of serine/threonine kinases, act as molecular switches in cytoskeletal-dependent processes involved in cellular motility. We hypothesized that PAK1 modulated eosinophil infiltration in an allergic airway disease (AAD) murine model. In this model, Pak1 deficient mice developed reduced inflammatory AAD responses in vivo with notable decreases in eosinophil infiltration in the lungs and broncho-alveolar lavage fluids (BALF). To test the importance of PAK1 in hematopoietic cells in AAD we used complementary bone marrow transplant experiments that demonstrated decreased eosinophil inflammation in hosts transplanted with Pak1 deficient bone marrow. In in vitro studies, we show that eotaxin-signaling through PAK1 facilitated eotaxin-mediated eosinophil migration. Ablating PAK1 expression by genetic deletion in hematopoietic progenitors or siRNA treatment in derived human eosinophils impaired eotaxin-mediated eosinophil migration, while ectopic PAK1 expression promoted this migration. Together these data suggest a key role for PAK1 in the development of atopic eosinophil inflammation and eotaxin-mediated eosinophil migration.

The allergic disease of asthma is characterized by an infiltration of inflammatory cells to the lung, a process co-ordinated by T-helper (TH) cells. The TH2 cytokine Interleukin (IL)-4 promotes infiltration of eosinophils to sites of inflammation. Eosinophil-selective chemoattractant cytokines (eg. eotaxins) are synthesized by lung epithelial cells. Eotaxin-3 is expressed at high levels in the asthmatic lung, predominantly after IL-4 stimulation. Eotaxin-3 is therefore a marker of inappropriate airway inflammation. Polyphenolic (PP) compounds found in high concentrations in berries may have beneficial effects in inflammatory conditions. Plant and Food Research produced high-PP extracts of blackcurrant (BC) cultivars that were tested for inflammation modulating effects. Since high doses of PPs have been shown to cause cell death, we tested two BC cultivars at a range of concentrations in a cell viability (WST-1) assay. While no toxic effects were attributable to the BC extracts (1-50µg/ml), a dose-related trend in cell death was observed and therefore 10µg/ml was chosen for further experiments Ten BC cultivars were compared for efficacy by measuring eotaxin-3 production in IL-4 stimulated human lung epithelial (A549) cells in vitro. Cells were incubated with BC extracts (10µg/ml) and IL-4 (10ng/ml) for 24 hours. The supernatants were then quantified for eotaxin-3 levels by an enzyme-linked immunosorbent assay (ELISA). All ten BC extracts reduced eotaxin-3 levels after stimulation with IL-4, and six BC extracts were effective by statistically significant levels (P<0.05), (BC cultivars -01, -02, -03, -05, -09 & -10). Of those, BC extracts of four cultivars demonstrated a reduction of more than 65% from the IL-4 stimulated control. In addition, a positive trend in inflammation modulation vs. one anthocyanin (ACN) in the BC extracts was shown. This study has demonstrated the beneficial inflammation modulatory effects of polyphenolic BC extracts, which could be related to cyanidin 3-O-rutinoside content. These results may have therapeutic potential for asthma.