Identification and classification of distinct surface markers of T regulatory cells.

Background: Regulatory T (Treg) cells have emerged as key players in the maintenance of immune homeostasis. Although significant progress has been made in recent years to define the Treg surface markers involved with or identifying their suppressive function, there remains much to be elucidated, and many questions persist. This study determined the expression of surface markers on human peripheral Treg cells and conventional T (Tconv) cells in a steady state and after activation to gain insight into their mechanism of action and more precisely characterize this regulatory population in humans. Methods: To screen Treg and Tconv cells, peripheral blood mononuclear cells (PBMCs) were isolated from volunteers, stained with a commercially available lyophilized antibody array comprising 371 surface antigens, and analyzed by flow cytometry. To compare Treg cells with activated Tconv cells, PBMCs were stimulated with PMA and further stained similar to freshly isolated cells. Results: Treg and Tconv cells were positive for 135 and 168 of the 371 antigens, respectively. Based on the frequency distribution, all of the most highly expressed markers identified were shared by both Treg and Tconv cells and participate in T cell activation, act as costimulatory and signaling molecules, or exhibit adhesion and migratory functions. Additionally, we identified several differences in marker expression between Treg and Tconv cells, with most found in the expression of co-stimulatory (ICOS, GITR, 4-1BB) and co-inhibitory (TIGIT, CTLA-4) molecules, as well as chemokine receptors (CXCR4, CXCR5, CCR4, CCR5, CCR7, CCR8, and CXCR7). Furthermore, post-activation expression of surface molecules identified molecules capable of discriminating Treg cells from activated Tconv cells (GITR, 4-1BB, TIGIT, CD120b, and CD39); however, almost all of these markers were also expressed in a small fraction of activated Tconv cells. Conclusions: These results offer insight into the biology of Tregs and contribute to their accurate identification and characterization in variety of immunological diseases as well as physiological processes.

Assessment of airway remodeling by quantitative computed tomography at various degrees of asthma severity defined according to the Global Initiative for Asthma report: a single-center study.

INTRODUCTION: In asthma, airway remodeling is defined as structural changes of the airways. The association between remodeling and asthma severity is still unclear, and there are limited data on the intensity of airway remodeling in various stages of the disease as defined in the Global Initiative for Asthma (GINA) asthma severity classification. Computed tomography (CT) and postprocessing applications are effective tools to assess the intensity of airway remodeling. OBJECTIVES: The aim of this study was to assess the severity of morphometric abnormalities of the respiratory tract in patients with various degrees of asthma severity according to the GINA guidelines. PATIENTS AND METHODS: The study included 70 patients with asthma and 29 healthy controls matched for age, sex, and body mass index. Patients were examined with a 128 multislice CT scanner at full inspiration. The measurements were made from the third to the ninth generations of bronchi. Bronchial parameters were compared between patients with severe and nonsevere asthma and healthy controls. RESULTS: We found no differences in the thickness of the bronchial wall, percentage of the wall area, inner and outer bronchial diameters, and the size of the bronchial lumen between severe and nonsevere asthma groups. Significant differences were noted in the thickness of the bronchial wall and the percentage of the wall area between the severe asthma group and the control group (P <0.017) as well as between the nonsevere asthma group and controls (P <0.017). CONCLUSIONS: Our findings indicated similar values of CT morphological measures of airway remodeling in all asthma severity groups as defined by the GINA guidelines.

Similarities in Quantitative Computed Tomography Imaging of the Lung in Severe Asthma with Persistent Airflow Limitation and Chronic Obstructive Pulmonary Disease.

BACKGROUND: Severe asthma with persistent airflow limitation (SA-PAL) and chronic obstructive pulmonary disease (COPD) are characterised by irreversible airflow limitation and the remodelling of the airways. The phenotypes of the diseases overlap and may cause diagnostic and therapeutic concerns. METHODS: There were 10 patients with SA-PAL, 11 patients with COPD, and 10 healthy volunteers (HV) enrolled in this study. The patients were examined with a 128-multislice scanner at full inspiration. Measurements were taken from the third to ninth bronchial generations. RESULTS: The thickness of the bronchial wall was greater in the SA-PAL than in the COPD group for most bronchial generations (p < 0.05). The mean lung density was the lowest in the SA-PAL group (-846 HU), followed by the COPD group (-836 HU), with no statistical difference between these two groups. The low-attenuation volume percentage (LAV% < -950 HU) was significantly higher in the SA-PAL group (15.8%) and COPD group (10.4%) compared with the HV group (7%) (p = 0.03). CONCLUSION: Severe asthma with persistent airflow limitation and COPD become similar with time within the functional and morphological dimensions. Emphysema qualities are present in COPD and in SA-PAL patients.

Quantitative Assessment of the Airway Response to Bronchial Tests Based on a Spirometric Curve Shift.

OBJECTIVE: Although spirometry is the most common pulmonary function test, there is no method to quantitatively infer about airway resistance or other properties from the flow-volume curves. Recently, an identifiable inverse model for forced expiration was proposed, as well as the idea to deduce changes in airway resistances and compliances from spirometric curve evolution. The aim of this work was to combine the above advances in a method for assessing the airway response to bronchial tests from a spirometric curve shift. METHODS: The approach is based on the differential measurement of the degree, site of maximal effect and width of changes, further recalculated into relative changes in the distribution of airway resistances (δRg) and compliances (δCg) along the bronchial tree. To this end, appropriate models were identified using the pre- and post-test spirometry data. The accuracy was validated using sets of data simulated by the anatomy and physiology based models. Finally, the method was used to analyze the bronchodilation tests of three asthmatic subjects. RESULTS: The expected errors in assessing the degree, site and width of changes in the zone of conducting airways were 6.3%, 2.4 generations and 22%, respectively, and for δRg and δCg were 5-10% and 13-16%, respectively. The analyses of clinical data indicated a significant reduction in resistances and an increase in compliances of airway generations 8-12, consistent with clinical knowledge. CONCLUSION: An unprecedented method to plausibly transforming the spirometry data into the site and degree of changes in airway properties has been proposed. SIGNIFICANCE: The method can be used to deduce about the effects of bronchial tests, as well as to monitor changes in the airways between visits or to investigate how inhaled pharmaceuticals affect the bronchi.

Airway wall thickness and airflow limitations in asthma assessed in quantitative computed tomography.

BACKGROUND: Asthma is a frequent chronic disease of the airways. In spite of the fact that symptoms of asthma are well known, the pathogenesis has not yet been fully understood. Quantitative computed tomography (qCT) of the lung allows for the measurment of a set of parameters. The aim of this study was to evaluate the usefulness of quantitative computed tomography in the assessment of airway wall thickness in asthma. METHODS: The prospective study was performed on a group of 83 patients with well-defined, long-term asthma between 2016 and 2018. The control group was composed of 30 healthy volunteers. All examined subjects were non-smokers. All computed tomography (CT) studies were performed using a 128 multi-slice CT scanner with no contrast, following a chest scanning protocol in the supine position, at full inspiration and breath-holds. RESULTS: Quantitative bronchial tree measurements were obtained from the third up to the ninth generation of the posterior basal bronchi (B10) of the right lung in a blinded fashion. The value of the wall thickness in patients with asthma was significantly higher in all measured generations of the bronchial tree (third to ninth generation). The lumen area and the inner diameter significantly correlated with the lung function tests and were substantially smaller in the examined group from the seventh to the ninth generation of the bronchi (p < 0.05). CONCLUSIONS: We conclude that airway remodelling occurs in most patients with long-term asthma and is associated mainly with the medium and small airways. Imaging techniques, especially qCT can be useful in the diagnosis and management of asthma. The reviews of this paper are available via the supplemental material section.

Asthma-COPD Overlap-A Discordance Between Patient Populations Defined by Different Diagnostic Criteria.

BACKGROUND: The concordance between asthma-chronic obstructive pulmonary disease overlap (ACO) defined according to Global Inititative for Asthma (GINA)/Global Initiative for Chronic Obstructive Lung Disease (GOLD) and other diagnostic criteria is unknown. OBJECTIVE: To assess the concordance between different ACO definitions and to estimate the definition-based ACO prevalence and characteristics. METHODS: A prospective, real-life study based on a 32-item data set was performed in a mixed population of patients with asthma and chronic obstructive pulmonary disease (COPD). Five different definitions of ACO, including the GINA/GOLD criteria, were analyzed. RESULTS: A total of 1609 patients were included in the final analysis. Application of Venn diagram for ACO populations resulted in 31 ACO subpopulations, which were further reduced to 6 separate populations by introducing a rank order for the analyzed definitions to classify patients from intersecting groups. Overall, the level of agreement between different ACO definitions was poor. Cohen kappa coefficient for the agreement between ACO GINA/GOLD definition and other ACO definitions varied from 0.06 to 0.21. Only 2 patients (0.12%) met all the ACO definitions. Definition-based ACO prevalence ranged between 3.8% (Spanish criteria) and 18.4% (clinician's diagnosis). A total of 573 (33.4%) patients met the criteria from at least 1 ACO definition. Patients who could not be classified as suffering from "pure" asthma, "pure" COPD, or ACO accounted for as much as 27.5% of the whole investigated group. The most severe symptoms were observed in patients with ACO defined as COPD and asthma diagnosed at age less than 40 years. CONCLUSIONS: The current ACO definitions identify distinct populations that share only a small number of common features and present with different disease phenotypes. ACO prevalence is highly variable, depending on the definition applied.

Autoreactive IgE in Chronic Spontaneous/Idiopathic Urticaria and Basophil/Mastocyte Priming Phenomenon, as a Feature of Autoimmune Nature of the Syndrome.

Recent years of research have shed a new light on the role of IgE in immune reactions. It seems to be more than just a contribution to immediate type of allergic response. It appears that monomeric IgE may enhance mast cell activity without cross-linking of FcεRI by IgE specific allergen or autoreactive IgG anti-IgE antibodies. Monomeric IgE molecules are heterogeneous concerning their ability to induce survival and activation of mast cells only by binding the IgE to FcεRI, but not affecting degranulation of cells. It also turned out that IgE may react to autoantigens occurring in the blood not only in chronic spontaneous urticaria (CSU) but also in other autoimmune diseases. The aforementioned phenomena may promote the activity of mast cells/basophils in CSU that easily degranulate when influenced by various inner (autoreactive IgG against IgE and FcεRI, autoreactive IgE for self-antigens) and outer factors (cold, heat, pressure) or allergens. These findings forced the new approach to the role of autoimmunity, self-antigens and IgE autoantibodies in the pathology of CSU. CSU put in the scheme of autoreactive IgG and autoreactive IgE seems to be either a kind of an autoimmune disease or a clinical manifestation of some other defined autoimmune diseases or both.

Impact of Birth Weight and Smoking on Lung Function in Patients with Asthma, COPD, and Healthy Volunteers.

BACKGROUND: Birth weight (BW) is an important factor for determining the development of the respiratory system. The majority of research analyzed the impact of BW on lung function in youth. BW influence and smoking on lung function in adults with asthma and COPD is an interesting issue. OBJECTIVES: The aim of the study was to investigate relationships between BW, smoking, and lung function in adult healthy individuals and diagnosed with asthma or COPD. MATERIAL AND METHODS: Four hundred seventy-nine subjects were divided into 5 groups: 123 healthy non-smokers, 180 healthy smokers, 72 non-smoking asthmatics, 57 smoking asthmatics, and 47 COPD patients. Relationships between 4 BW quartiles and lung function was analyzed with respect to smoking. RESULTS: Impact analyzes of BW, smoking, and asthma on FVC% revealed that asthma is the only significant differentiating factor in this spirometric parameter (p < 0.01). FEV1% was significantly influenced by asthma and BW, and FEV1/ FVC% was exclusively influenced by asthma. Spirometric parameters increased proportionally to particular BW quartiles in healthy non-smokers group; however optimal BW quartile predicting increase of parameters was 2751-3250 g. In asthma, BW quartile predicting the increase of spirometric parameters was 3251-3750 g, but BW quartile predicting decrease of FEV1/FVC% was 2751-3250 g. The comparison of results between COPD group and results from other 4 groups showed that values of all parameters in patients with COPD did not change proportionally to all quartiles of BW. In terms of FEV1/FVC%, the proportional increase of parameter in BW quartile 2751-3250 g was observed. CONCLUSIONS: BW, as independent factor influences on spirometric parameters of healthy individuals, patients with asthma, COPD in a differentiated manner depending on quartile of BW rather than on simple linear increase of BW, regardless of smoking.

Airway Evaluation with Multidetector Computed Tomography Post-Processing Methods in Asthmatic Patients.

Asthma is a chronic inflammatory obstructive airways disease. The disease occurs regardless of age and manifests with cough, attacks of breathlessness, and tightness in the chest. The pathophysiology of asthma is complex and still not fully understood. It is essential to find answers concerning the role of each part of the bronchial tree in asthma, especially the role of small bronchioles. With the development of newer generations of multidetector computed tomography (MDCT) and advanced post-processing methods it is possible to obtain more detailed images and gain insight into further aspects of asthma. MDCT post-processing methods can be divided into two-dimensional (2D) and three-dimensional (3D). In 2D projections, visualized hypodense regions correspond to the airway flow limitations. With the more advanced methods, such as multi planar reconstructions (MPR), images in different planes (axial, coronal, or sagittal) can be created. In the MPR technique only the voxels which are adjacent to each other in the predetermined plane can be extracted from the data set. Using the minimal/maximal intensity projections and shaded surface display, the volume of interest (VOI) can be extracted. High resolution CT scans can be used to create a more advanced imaging tool - the virtual bronchoscopy (VB). Using the VB makes it possible to visualize regions of obturation in the bronchi of up to the 5-8th generation. The MDCT with advanced post-processing methods is likely to assume an important role in the differential diagnosis of asthma, particularly when the diagnosis is dubious or hard to settle due to accompanying other lung diseases.

Hospital management of patients with exacerbation of severe chronic obstructive pulmonary disease.

The article assesses the originally developed criteria of clinical stability and treatment protocol in the hospital management and discharge procedures of patients with exacerbations of severe chronic obstructive pulmonary disease (COPD). The study included 34 patients (26 males, 8 females), aged 58-80 years, hospitalized due to exacerbation of severe (23 patients) and very severe (11 patients) COPD. On admission, the mean FEV1 was 0.78 ± 0.22 L (31.7% ± 8.2% of predicted), FVC 2.52 ± 0.87 L (77.9% ± 9.8% of predicted) and FEV1/FVC 33.17% ± 10.84%. Before hospitalization, 10 out of the 34 patients were diagnosed with chronic respiratory failure. All patients were treated according the same treatment protocol which included the developed criteria of clinical stability. Meeting all these criteria in a 24-h observation period was the basis to slash the dose of systemic glucocorticosteroids by half. The maintenance of the stability criteria through the subsequent 24 h allowed discharging a patient from the hospital. Every patient was supplied with a detailed plan of out-of-hospital treatment. The results show that the mean duration of hospitalization was 6.4 ± 4.8 days. Only one patient required readmission within 4 weeks after discharge. Two patients died; one during the hospitalization time and the other after discharge. In the latter case, death was not directly related to the COPD exacerbation. In conclusion, the protocol of treatment and the criteria of stability used for patients with COPD exacerbation enabled to optimize the hospitalization time. A shortening of hospitalization was not associated with increased risk of readmission within 4 weeks after discharge.

Polymorphism of the TGFB1 gene is not associated with bronchial allergic asthma in a Polish population.

Allergic asthma is a complex genetic disorder that involves interactions between genetic and environmental factors. Some studies have indicated that transforming growth factor beta(1) (TGF-beta(1)), a pleiotropic cytokine regulating inflammatory reactions and airway remodeling, may participate in the pathogenesis of asthma. Several polymorphisms have been described in the TGFB1 gene; some were tested in allergic asthma, with conflicting results. The aim of this study was to investigate the possible associations of four TGFB1 gene polymorphisms (-800G>A, -509C>T, 869T>C, and 915G>C) with allergic asthma in a Polish population. These four single nucleotide polymorphisms were genotyped in 247 asthmatic patients (including 207 atopic individuals) and 287 unrelated healthy volunteers by means of the polymerase chain reaction-restriction fragment length polymorphism method. No significant differences between patients and controls in allele, genotype, and haplotype frequencies were reported. Logistic regression analysis of genotype distribution and allele positivity adjusted for age and sex did not reveal any significant differences between all patients or patients selected for atopy and controls. Thus, no evidence was reported for a contribution of the TGFB1 gene to allergic asthma in a Polish population. The results are discussed in the context of similar studies in other populations.

Association of PTPN22 single nucleotide polymorphism with rheumatoid arthritis but not with allergic asthma.

PTPN22 gene encodes a lymphoid tyrosine phosphatase (LYP), an important negative regulator of T-cell responses. The 1858C>T (Arg620Trp) single nucleotide polymorphism (rs2476601) was found associated with autoimmune diseases, including rheumatoid arthritis (RA). Allergic diseases are similar to autoimmune diseases, by an exaggerated immune response to an antigen (allergen in this case) normally not invoking such response in healthy individuals. We investigated whether polymorphism 1858C>T in PTPN22 gene is associated with susceptibility to allergic asthma and RA in a Polish population. PTPN22 was genotyped in 173 patients with RA, in 198 patients with allergic asthma, and in 543 controls using PCR-RFLP. The patients with RA differed from healthy controls in frequencies of PTPN22 1858C>T alleles (P=0.0004; odds ratio (OR), 1.8; 95% CI, 1.33-2.55) and genotypes (P=0.0009). Strong associations of 1858T allele with RA limited to joints (0.21 vs 0.12, P=0.0002; OR, 2.1; 95% CI, 1.44-3.00), with erosive disease (0.20 vs 0.12, P=0.0003; OR, 1.92; 95% CI, 1.34-2.71), with a lack of rheumatoid factor (RF; 0.23 vs 0.12, P=0.0008; OR, 2.29; 95% CI, 1.44-3.63), and weak association with the presence of RF (0.17 vs 0.12, P=0.02; OR, 1.6; 95% CI, 1.10-2.40) in comparison with healthy controls were observed. Very strong association of 1858T allele (P<0.0001; OR, 2.72; 95% CI, 1.9-3.9) and T phenotype (P<0001; OR, 3.2; 95% CI, 2.1-4.9) with antibodies to cyclic citrullinated peptide (CCP) was found. When patients with allergic asthma were typed for PTPN22 1858C>T polymorphism, no difference with control was found. Subdivision of patients into those with mild, moderate, or severe asthma did not reveal any associations. In conclusion, we confirmed associations between several clinical manifestations of RA and PTPN22 1858T allele. However, no association with 1858C>T polymorphism was found for susceptibility to allergic asthma or for severity of the disease.

The use of magnesium in bronchial asthma: a new approach to an old problem.

Magnesium deficiency is a common electrolyte disorder in patients with acute severe asthma, but intracellular magnesium content better reflects its homeostasis than does its serum concentration. Magnesium takes part in many metabolic processes in the organism, including energy metabolism, protein and nucleic acid synthesis, cell cycle, the binding of substances to the plasma membrane, and maintenance of cytoskeletal and mitochondrial integrity. It also modulates ion transport and influences intracellular calcium concentration. Maintenance of the cells' transmembrane gradient depends on the presence of magnesium, and hypomagnesemia may result in an increase in neuromuscular cell excitability. Magnesium is a cation modulating the smooth muscle contractility of different tissues: hypomagnesemia causes their contraction and hypermagnesemia their relaxation. Suggestions of a positive influence of magnesium in the treatment of asthma exacerbation have been known for a long time, but research results differ. A single dose of intravenous magnesium sulfate given to patients with acute asthma exacerbation has been shown to be safe, but its efficiency is still under discussion. According to the Global Initiative for Asthma GINA-2005, magnesium sulfate administration is not recommended for routine treatment, but it is permitted in patients with severe asthma exacerbation not responding to treatment (evidence category A). Recommendations of the British Thoracic Society allow one dose of magnesium sulfate to patients with acute severe asthma exacerbation and inadequate initial response to broncho-dilating inhalation treatment (evidence category A). Future investigations should help to establish the indications for magnesium use in the treatment of acute asthma exacerbations as well as the magnesium dose and the scheme of its administration.

Distribution of CTLA-4 polymorphisms in allergic asthma.

BACKGROUND: The CTLA-4 molecule is an important negative regulator of T cell activation. It is encoded on chromosome 2q33 and found to be associated with several allergic phenotypes including asthma. However, the association of CTLA-4 gene polymorphisms with allergic asthma is still controversial and therefore was the subject of this study. METHODS: By PCR-RFLP, the distribution of three single nucleotide polymorphisms (SNPs), -1147 C/T, -318 C/T, and +49 A/G, was examined in 219 Polish Caucasoid patients diagnosed with allergic asthma and in 102 ethnically matched healthy control individuals. (AT)(n) microsatellite polymorphism was also tested in the same individuals. RESULTS: No statistically significant differences in SNPs or microsatellite allele, genotype or haplotype frequencies between patients and controls were found. CONCLUSION: CTLA-4 polymorphisms do not seem to be a risk factor for allergic asthma in Poles.

Are single nucleotide polymorphisms of the immunoglobulin A Fc receptor gene associated with allergic asthma?

BACKGROUND: Eosinophils are important components of allergic inflammation. The immunoglobulin A (IgA) Fc receptor (FcalphaRI), encoded by the FCAR gene, is a possible candidate for eosinophil activation at mucosal surfaces, where IgA is abundant. Both elevated cell surface expression of FcalphaRI and increased avidity for IgA were described on eosinophils from allergic subjects. The aim of our study was to examine the possible association of FCAR gene polymorphisms with allergic asthma. METHODS: We screened three regions of the FCAR gene: (1) the promoter region, (2) exon 3, encoding the first extracellular domain (EC1), and (3) exon 5, coding for the transmembrane and cytoplasmic domain, for new and published polymorphisms using a sensitive temperature gradient gel electrophoresis technique and compared their frequencies in 112 patients diagnosed with allergic asthma and 100 healthy controls. RESULTS: Six polymorphisms, including two novel ones, were detected. No differences between patients and controls were found in the distribution of any of these polymorphisms. CONCLUSION: FcalphaRI polymorphism does not seem to be a risk factor in allergic asthma. Nevertheless, this is the first report on the distribution of 6 single nucleotide polymorphisms of the FCAR gene in a human population and the first study on FCAR polymorphism in allergic asthma.

Prevalence of Hymenoptera venom allergy and its immunological markers current in adults in Poland.

BACKGROUND: The prevalence of insect venom allergy is still being assessed. The aim of our study was to estimate, on the basis of an interviewer-administered questionnaire survey, the frequency of post-sting allergic reactions and venom sensitization. MATERIAL/METHODS: The study was performed within the framework of the ECRHS. A random sampling of 3000 persons was selected from among 68,000 persons living in the area of Wroclaw, Poland. Of the 2050 persons responding to a mailed screening questionnaire, 169 were randomly selected to complete a questionnaire designed only for insect allergy detection. Venom skin test and sIgE assessment were performed on 146 and 132 patients, respectively. RESULTS: Allergic post-sting symptoms were found in 20.7% of surveyed patients. Large local reactions (LLs) occurred in 11.8% and systemic reactions (SYSs) in 8.9% of the study population. SYS was most often manifested by urticaria (4.7%). The frequencies of SYS II, III and IV were 1.8%, 1.8%, and 0.6%, respectively. Only LLs were more frequent in subjects with other allergic diseases (p=0.03). The presence of positive skin tests and/or sIgE in serum were 42.8% of subjects with LL, 53.3% with SYS, and 17.1% of "asymptomatic" patients. No significant differences were found between these groups regarding venom skin test results and sIgE serum concentrations. Occurrence of sIgE to bee venom was frequently associated with the presence of sIgE to timothy grass. CONCLUSIONS: Insect venom allergy and asymptomatic venom sensitization in adults are common in Poland. Only some venom allergy cases are IgE dependent.

[Inhaled glucocorticosteroids in the treatment of asthma and chronic obstructive pulmonary disease]. / Glikokortykosteroidy wziewne w leczeniu astmy oskrzelowej i przewleklej obturacyjnej choroby pluc.

Molecular mechanisms of actions, pharmacokinetics and anti-inflammatory potency of inhaled glucocorticosteroids (ICS) are described. Differences in clinical effectiveness of ICS in asthma and chronic obstructive pulmonary disease (COPD) therapy and up-to-date recommendations for treatment with ICS in asthma and COPD patients are discussed. Finally there are presented data on safety of long-term treatment with ICS, specially in asthmatic children.