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Atopic dermatitis is a chronic condition of complex etiology, whose clinical course involves remission and recurrence. It is not an isolated disease entity affecting only the skin, but one that co-occurs with disorders of other organs. Numerous literature reports have long confirmed the relationship between the disorder and a growing number of ophthalmic manifestations such as keratoconus and retinal detachment. Further studies are required to establish the cause of correlations and to allow for implementation of appropriate prophylaxis and treatment. The aim of the present paper is to review published literature regarding the correlation between atopic dermatitis and ophthalmic manifestations in adults and children.

To the editor,Atopic dermatitis (AD) is a common, chronic disease with complex etiology affecting approximately up to 20% of children and skin barrier dysfunction plays a pivotal role in its pathog...

BACKGROUND: It is unclear whether asthma may affect susceptibility or severity of coronavirus disease 2019 (COVID-19) in children and how pediatric asthma services worldwide have responded to the pandemic. OBJECTIVE: To describe the impact of the COVID-19 pandemic on pediatric asthma services and on disease burden in their patients. METHODS: An online survey was sent to members of the Pediatric Asthma in Real Life think tank and the World Allergy Organization Pediatric Asthma Committee. It included questions on service provision, disease burden, and the clinical course of confirmed cases of COVID-19 infection among children with asthma. RESULTS: Ninety-one respondents, caring for an estimated population of more than 133,000 children with asthma, completed the survey. COVID-19 significantly impacted pediatric asthma services: 39% ceased physical appointments, 47% stopped accepting new patients, and 75% limited patients' visits. Consultations were almost halved to a median of 20 (interquartile range, 10-25) patients per week. Virtual clinics and helplines were launched in most centers. Better than expected disease control was reported in 20% (10%-40%) of patients, whereas control was negatively affected in only 10% (7.5%-12.5%). Adherence also appeared to increase. Only 15 confirmed cases of COVID-19 were reported among the population; the estimated incidence is not apparently different from the reports of general pediatric cohorts. CONCLUSIONS: Children with asthma do not appear to be disproportionately affected by COVID-19. Outcomes may even have improved, possibly through increased adherence and/or reduced exposures. Clinical services have rapidly responded to the pandemic by limiting and replacing physical appointments with virtual encounters. (C) 2020 American Academy of Allergy, Asthma & Immunology.

BACKGROUND Low 25-hydroxyvitamin D (25[OH]D) and asthma development may be related to airway remodeling and eosinophilia. Periostin is proposed as a key molecule that links remodeling and eosinophilic inflammation. OBJECTIVE We evaluated the association of 25(OH)D concentration with periostin, peripheral blood eosinophil counts, and immunoglobulin E (IgE) in children with newly diagnosed asthma. METHODS The study included 150 children: 110 with atopic asthma and 40 constituted a reference group. Fasting blood was collected for cell counts and serum for measurements of 25(OH)D, periostin, IgE, and C-reactive protein (CRP) concentrations. RESULTS Significantly lower 25(OH)D, elevated IgE concentrations, and eosinophil counts were found in children with asthma compared with the reference group (p = 0.0001). A lower forced expiratory volume in the first second of expiration percentage predicted value was associated with a lower 25(OH)D value in children with asthma. The bronchodilator reversibility was inversely related to serum 25(OH)D concentrations (R = -0.45, p = 0.029). The children with asthma and with a 25(OH)D deficient concentration (≤20 ng/mL) had higher concentrations of periostin (p = 0.035) and CRP (p = 0.01) than those with a sufficient 25(OH)D concentration (≥30 ng/L). Additional analysis revealed statistically significant differences (p = 0.013) when comparing periostin concentrations between subjects with a 25(OH)D deficient concentration (≤20 ng/mL) and subjects who did not have a deficient concentration (>20 ng/mL). In individuals with asthma, a 25(OH)D concentration of

Julian F Guest,1,2 Diana Weidlich,1 Maciej Kaczmarski,3 Elzbieta Jarocka-Cyrta,4 Natalia Kobelska-Dubiel,5 Agnieszka Krauze,6 Iwona Sakowska-Maliszewska,7 Anna Zawadzka-Krajewska8 1Catalyst Health Economics Consultants, Northwood, Middlesex, 2Faculty of Life Sciences and Medicine, King’s College, London, UK; 3Department of Pediatrics, Gastroenterology, and Allergology, Medical University of Bialystok, Białystok, 4Uniwersytet Warmińsko‑Mazurski, Wydział Nauk Medycznych, Katedra Pediatrii Klinicznej, Olsztyn, 5Department of Pediatric Gastroenterology and Metabolic Diseases, Poznań University of Medical Sciences, Poznań, 6Klinika Pneumonologii i Alergologii Wieku Dziecięcego, Warsaw, 7Poradnia Gastroenterologiczna Centrum Pediatrii, Sosnowiec, 8Department of Pediatric Pneumonology and Allergology, Medical University of Warsaw, Warsaw, Poland Objective: To estimate the cost-effectiveness of using an extensively hydrolyzed casein formula (eHCF) containing the probiotic Lactobacillus rhamnosus GG (eHCF + LGG; Nutramigen LGG) as an initial treatment for cow’s milk allergy compared with eHCF alone and amino acid formulas (AAF) in Poland from the perspective of the Polish National Health Fund (Narodowy Fundusz Zdrowia [NFZ]) and parents. Methods: Decision modeling was used to estimate the probability of cow’s milk allergic infants developing tolerance to cow’s milk by 18 months. The model also estimated the cost to the NFZ and parents (Polish Zloty [PLN] at 2013–2014 prices) for managing infants over 18 months after starting one of the formulas as well as the relative cost-effectiveness of each of the formulas. Results: The probability of developing tolerance to cow’s milk by 18 months was higher among infants who were fed eHCF + LGG (0.82) compared with those fed eHCF alone (0.53) or an AAF (0.22). An infant who is initially managed with eHCF + LGG is expected to consume fewer health care resources than infants managed with the other formulas. Hence, the estimated total health care cost incurred by the NFZ for initially feeding infants with eHCF + LGG (PLN 5,693) was less than that of feeding infants with eHCF alone (PLN 7,749) or an AAF (PLN 24,333). However, the total cost incurred by parents for initially feeding infants with an AAF (PLN 815) was marginally less than that of feeding with eHCF + LGG (PLN 993), which was less than that of feeding with eHCF alone (PLN 1,226). Conclusion: Using eHCF + LGG instead of eHCF alone or an AAF for first-line management of newly diagnosed infants with cow’s milk allergy affords a cost-effective use of NFZ-funded resources, since it improves outcome for less cost. Whether eHCF + LGG would be viewed as being cost-effective by parents is dependent on their willingness to pay an additional cost for additional tolerance acquisition to cow’s milk. Keywords: amino acid formula, cost-effectiveness, cow’s milk allergy, extensively hydrolyzed formula, Lactobacillus Rhamnosus GG, Poland

Wstep: Kliniczna skutecznośc immunoterapii podjezykowej zostala juz gruntowanie potwierdzona i udokumentowana. Badano rowniez jej wplyw na uklad odpornościowy chorych na astme oskrzelową. J ednak do tej pory nie wyjaśniono zalezności pomiedzy wystepowaniem alergii wielowaznej, wspolwystepowaniem innych chorob alergicznych i stopniem terapii astmy a wplywem prowadzonej alergenowo-swoistej immunoterapii podjezykowej. Celem niniejszej pracy byla ocena wplywu rocznej immunoterapii podjezykowej na wybrane parametry odpowiedzi immunologicznej w zalezności od cech osobniczych i klinicznych cech choroby u pacjentow chorych na astme i alergiczny niezyt nosa. Material i metody: Do badania zakwalifikowano 25 dzieci w wieku 8,1 ± 3,1 roku (5–15 lat), 21 chlopcow i 4 dziewczynki, chorych na astme oskrzelową i alergiczny niezyt nosa z alergiami poliwalentnymi sezonowymi i niesezonowymi, ktore byly zakwalifikowane do alergenowo-swoistej immunoterapii podjezykowej. Odsetek limfocytow pomocniczych Th1 i Th2, ekspresje antygenu Bcl-2 w limfocytach T oraz aktywacje bazofilow pod wplywem swoistych alergenow (zawiesiny antygenow roztoczy kurzu domowego i/lub pylkow traw do testow skornych) badano metodą cytometrii przeplywowej. Z alezności pomiedzy badanymi parametrami oceniano dokladnym testem Fishera. Wyniki: Nie zaobserwowano istotnych zalezności pomiedzy wplywem immunoterapii podjezykowej na uklad odpornościowy pacjentow a wiekiem, wystepowaniem alergii wielowaznej, stopniem terapii astmy i wspolwystepowaniem innych chorob alergicznych. Wzrost odsetka limfocytow pomocniczych Th1 pod wplywem immunoterapii podjezykowej obserwowano nieco cześciej u dzieci uczulonych na wiecej niz 3 alergeny niz u pozostalych. Wnioski: Na podstawie badan przeprowadzonych przez autorow pracy nie mozna wskazac zadnej grupy pacjentow, u ktorych mozna sie spodziewac silniejszej mobilizacji ukladu odpornościowego jako odpowiedzi na prowadzoną immunoterapie podjezykową u dzieci chorych na astme i alergiczny niezyt nosa. Wzrost odsetka limfocytow pomocniczych Th1 moze wystepowac cześciej u dzieci z alergią poliwalentną.

This article discusses the impact of changes in the respiratory system as a result of asthma on the function of respiratory muscles. It describes thoroughly the action of the respiratory muscles among healthy people and disorders that occur in the case of asthma. Hereinafter, it presents the correlation between the dysfunction of the respiratory muscles and body posture among children with asthma.

Objective: Allergen-specific immunotherapy (SIT) is the unique modifying treatment of atopic diseases. Dysregulation of T-cell apoptosis plays a crucial role in the in the development of asthma. Nevertheless, the effect of sublingual immunotherapy (SLIT) on T-cell apoptosis has not been elucidated. The aim of the study was to evaluate the influence of 1 year of SIT in atopic children on the frequency of Th1 and Th2 cells in peripheral blood, on T-cell apoptosis, and on the response of basophils to allergen challenge. Methods: Children suffering from bronchial asthma were treated with SLIT for 12 months (Staloral 300). Basophil activation was evaluated by measurement of CD203c antigen expression. Th1 and Th2 cells frequencies and their associated frequencies of apoptosis by the expression of Bcl-2 were evaluated with flow cytometry. Results: Basophil activation showed no difference in response before and after therapy. The frequency of Th1 cells increased (p=0.01, n=19), whereas the frequency of Th2 cells remained stable. Additionally, a significant increase of Bcl-2 positive Th1 cells was found (p=0.0465, n=19). Conclusions: We conclude that the increase in frequency of Th1 cells secondary to SLIT might be associated with increased resistance to apoptotic signals. The basophil activation is not useful in evaluation of patient desensitization.

Background: Atopic allergy is among the immune tolerance-related disorders resulting from a failure of the regulatory network. Regulatory T cells (Tregs) play a leading role in the development of homeostasis in the immune system. The aim of this study was to determine the role of Tregs in the pathogenesis of atopic diseases in children by exploring the relationship between Treg frequency, activation markers and the clinical manifestations of the disease. Methods: Twenty allergic and 50 healthy children were enrolled to the study. Peripheral blood mononuclear cells were stained with monoclonal antibodies (anti-CD25-CD4-CD127-FoxP3-CD69-CD71) and evaluated using flow cytometry. Tregs were identified as CD4+CD25+/highFoxP3+CD127- T cells. Results: The percentage of Tregs in allergic patients (2.3%) was significantly decreased in comparison to healthy controls (4.6%, p = 0.003). The frequency of Tregs in patients with symptoms of atopic dermatitis and/or food allergy (1.7%) was significantly lower than in patients without these symptoms (2.9%, p = 0.04). A significant correlation between the percentage of Tregs and the sIgE serum concentration was observed (p = 0.037). Relative fluorescence intensities of FoxP3 expression in allergic patients were higher than in healthy controls (p = 0.00004). The frequency of CD4+CD25highCD127-CD71+ cells did not differ between the groups. Conclusions: Tregs display substantial deficiencies in atopic children, especially in children with multiorgan involvement, compared to patients with single organ manifestations. Additionally, there is an association between Tregs and the sIgE serum concentration. Better identification and characterization of Tregs in allergy is needed as they limit responses to foreign antigens, thereby minimizing T cell-mediated immunopathology in allergic diseases.

Objective To estimate the cost-effectiveness of using an extensively hydrolyzed casein formula (eHCF) containing the probiotic Lactobacillus rhamnosus GG (eHCF + LGG; Nutramigen LGG) as an initial treatment for cow’s milk allergy compared with eHCF alone and amino acid formulas (AAF) in Poland from the perspective of the Polish National Health Fund (Narodowy Fundusz Zdrowia [NFZ]) and parents. Methods Decision modeling was used to estimate the probability of cow’s milk allergic infants developing tolerance to cow’s milk by 18 months. The model also estimated the cost to the NFZ and parents (Polish Zloty [PLN] at 2013–2014 prices) for managing infants over 18 months after starting one of the formulas as well as the relative cost-effectiveness of each of the formulas. Results The probability of developing tolerance to cow’s milk by 18 months was higher among infants who were fed eHCF + LGG (0.82) compared with those fed eHCF alone (0.53) or an AAF (0.22). An infant who is initially managed with eHCF + LGG is expected to consume fewer health care resources than infants managed with the other formulas. Hence, the estimated total health care cost incurred by the NFZ for initially feeding infants with eHCF + LGG (PLN 5,693) was less than that of feeding infants with eHCF alone (PLN 7,749) or an AAF (PLN 24,333). However, the total cost incurred by parents for initially feeding infants with an AAF (PLN 815) was marginally less than that of feeding with eHCF + LGG (PLN 993), which was less than that of feeding with eHCF alone (PLN 1,226). Conclusion Using eHCF + LGG instead of eHCF alone or an AAF for first-line management of newly diagnosed infants with cow’s milk allergy affords a cost-effective use of NFZ-funded resources, since it improves outcome for less cost. Whether eHCF + LGG would be viewed as being cost-effective by parents is dependent on their willingness to pay an additional cost for additional tolerance acquisition to cow’s milk.

Background Detection of allergen-induced basophil activation by flow cytometry has been shown to be a useful tool for allergy diagnosis. Objective The aim of this study was to assess the potential of this technique for the diagnosis of pediatric cow milk allergy. Material and methods The quantification of total and specific IgE and basophil activation test were performed to evaluate cow milk allergic (n = 9), and non-allergic children (n = 15). Results Allergen-induced basophil activation was detected as a CD203c up-regulation. The expression of CD203c antigen on basophils was measured with flow cytometry. The antigen CD203c was detected on 15.4 ± 10.2% basophils from allergic children after incubation with specific allergens in concentration 1:10, whereas in the control group there were 3.0 ± 1.5% of basophils positive for this molecule (P < 0.05). Stimulation with allergen diluted 1:500 resulted in activation of 15.3 ± 11.2% of basophils in allergic children and 3.8 ± 2.3% of cells in the control group (P < 0.05). Positive results of an allergenicity test (above the cut- off level of 10%) were obtained in 7 out of the 9 allergic children. In 5 cases, the cutoff level was reached with both dilution of allergens (1:10 and 1:500). In 1 patient, positive stimulation was observed after stimulation with allergen diluted 1:10 and in another case only 1:500 resulted in stimulation of more then 10% of basophils. In no child of the control group, stimulation above 10% was noted. Conclusions This study demonstrates that the analysis of allergen-induced CD203c up-regulation with flow cytometry is a reliable tool for the diagnosis of cow milk allergy in pediatric patients, with sensitivity similar to routine diagnostic tests and a higher specificity.

The T cell hypothesis of asthma is based on the concept that the disease is driven and maintained by the persistence of a specialized subset of chronically activated T memory cells sensitized against an array of allergenic, occupational or viral antigens. Overreaction of CD4+ T cells in the peripheral blood and airway tissues is an invariant feature of asthma; therefore a potent mechanism for augmenting the number of activated T cells in this disease would be the resistance to the normally programmed pathway for cell death. The aim of the study was to evaluate the presence of apoptotic markers on peripheral blood lymphocytes from healthy and asthmatic children before and after stimulation with antiCD95 antibodies. The blood was collected from 21 children with atopic asthma suffering from allergic rhinitis because of house dust mite and/or grass pollen allergens and 8 healthy children matched for their age and sex. Blood was mixed with purified monoclonal antibody antiCD95 (Beckman Coulter), incubated for 24 hours and than stained with Annexin V andPI (Becton Dickinson). Prepared suspensions were analyzed with Cytomics FC 500 (Beckman Coulter) flow cytometer. Annexin V(+)/PI(-) cells were characterized as early apoptotic, Annexin V(+)/PI(+) as late apoptotic and Annexin V(-)/PI(+) as dead. In unstimulated sample from asthmatic children 21.09+/-11.20% cells were characterized as Annexin V positive/PI negative. After stimulation with antiCD95 Annexin V positive/PI negative cells constituted 18.72+/-9.42% of cells, p=0.1. In unstimulated sample from healthy children 11.69+/-6.70% cells were characterized as Annexin V positive/PI negative. In the sample stimulated with antiCD95 16.54+/-2.98% of cells were Annexin V positive/PI negative, p=0.02. There were no differences between results of late apoptotic and necrotic lymphocytes from healthy and asthmatic children. Performed research indicates that lymphocytes from asthmatic children are resistant to Fas mediated apoptosis.

Matrix metalloproteinase (MMP)-9 is involved in pathophysiology of asthma, mainly asthma-associated airway remodeling. Exhaled breath condensates (EBC) of asthmatics contain increased amounts of MMP-9 with activity higher, than in healthy controls. The increased activity of MMP-9 may originate from its excessive production and activation, but may also result from variations in MMP-9 structure, which are determined by single nucleotide polymorphisms (SNPs). In this pilot study we aimed to assess the possible influence of two functional MMP-9 polymorphisms, Q279R and R668Q, on enzymatic activity of MMP-9, measured in EBC of asthmatic children. The concentration and activity of MMP-9 were analyzed in EBC of 20 children with allergic asthma using specific standard ELISA and novel immunoenzymatic activity assay. The SNPs of MMP-9 were assessed using real-time PCR-based genotyping test. We have found that MMP-9 concentration in breath condensates of children with stable asthma was slightly higher in ELISA, than in the activity assay. Moreover, these results and activity-to-amount ratio have revealed some relationship with a presence of specific 279R and/or 668Q MMP-9 gene variants. Our observation suggests that at least in some patients MMP-9 hyperactivity may result from genetic predisposition, determined by polymorphic variants of MMP-9 gene. Moreover, it supports previous reports postulating significance of MMP-9 in pathogenesis of asthma. However, this issue still requires further studies.

Over the last decades allergic diseases has become a major health problem worldwide. The only specific treatment to date is allergen specific immunotherapy (ASIT). Although it was shown that ASIT generates allergen-tolerant T cells, detailed mechanism underlying its activity is still unclear and there is no reliable method to monitor its effectiveness. The aim of our study was to evaluate ASIT influence on the frequency of forkhead box P3 (FoxP3) Tregs in allergic children with various clinical manifestations. The relative number of FoxP3 Tregs in 32 blood samples from allergic children at baseline and/or after 1 year of ASIT was assessed by flow cytometry. In the entire studied group, the percentage of FoxP3 Tregs did not increase 1 year after ASIT. Nevertheless, the percentage of FoxP3 Tregs after ASIT significantly increased in children with respiratory allergy (conjunctivitis, asthma, and rhinitis) coexisting with nonrespiratory manifestations (food allergy and/or atopic dermatitis), whereas, in patients with respiratory allergy only, the percentage of FoxP3 Tregs decreased. To the best of our knowledge, this is the first report showing various differential FoxP3 Tregs response to ASIT in allergic children. FoxP3 Tregs number could be useful in treatment monitoring. Further studies are warranted to confirm these observations.

Exposure to environmental tobacco smoke (ETS) has been shown to increase symptoms of allergic bronchial asthma, but direct effects on the expression of inflammatory markers have not been demonstrated thus far.The aim of this study was to assess the correlation of ETS exposure with the expression of proinflammatory mediators in airway secretions, including IFN-gamma and IL-12, as well as IL-5 and IL-13, in allergic asthmatic schoolchildren and healthy control subjects.By using the nasopharyngeal aspiration technique, airway secretions were collected from 24 atopic children with asthma (age, 6-16 years) and 26 healthy control subjects, and the concentration of cytokines was measured with immunoenzymatic methods.IL-13 levels were highly increased in patients with asthma (P.005), and parental tobacco smoke resulted in a significant increase in airway IL-13 secretion in these children compared with that seen in nonexposed children and healthy control subjects (median, 860 pg/mL vs 242 pg/mL and 125 pg/mL, respectively). Furthermore, a positive correlation between IL-13 levels and serum IgE concentrations (r(s) = 0.55) was found in children with allergic asthma.These results indicate that ETS augments the expression and secretion of IL-13 in allergic asthma and that nasopharyngeal aspiration is a suitable method to assess cytokine measurements in airways in children. Measurements of IL-13 in secretions might be taken into account as a noninvasive marker of airway inflammation and to assess the detrimental effects of ETS.

In addition to Th2 cells, other T cell subsets, like Th1, Th17, and regulatory T cells are also involved in the pathogenesis of allergic rhinitis, food allergy, and other allergic diseases. The aim of the study was to evaluate a wide profile of cytokines in the plasma from children with pollen/dust and cow milk allergy with the use of a multiplex Cytometric Bead Array (CBA) test. Twenty children with allergy: 11 with pollen/dust and 9 with cow milk allergy were enrolled into the study. For the analysis of INFγ, IL-2, IL-4, IL-6, IL-10, IL-17A, and TNFα levels, a flow cytometric test was used. TGFβ concentration was measured using an ELISA test. Concentrations of almost all of the detected cytokines in the patient's plasma were below 1 pg/ml, with no significant difference between the groups examined. The level of IL-17A was higher in pollen/dust allergy group (median 12.73 pg/ml; 25th percentile-Q1/75th percentile-Q3; 9.28/15.83) in comparison to cow milk allergy group (median 7.86 pg/ml - Q1/Q3; 7.86/14.37). Children with cow milk allergy had a slightly higher concentration of TGFβ than children with pollen/dust allergy (median 28.7 ng/ml - Q1/Q3; 19.77/33.00 vs. 17.84 ng/ml - Q1/Q3; 11.21/23.75, respectively); the difference did not reach statistical significance. We conclude that plasma concentration of Th1, Th2, and Th17 regulatory cytokines are relatively low in allergic children. The CBA test, which could be useful in pediatric practice as it requires a relatively small plasma sample volume, is not sensitive enough to measure low cytokine levels, since its detection limit and the standard curve are not appropriate for the concentrations of these mediators expected in clinical samples.

In addition to Th2 cells, other T cell subsets, like Th1, Th17, and regulatory T cells are also involved in the pathogenesis of allergic rhinitis, food allergy, and other allergic diseases. The aim of the study was to evaluate a wide profile of cytokines in the plasma from children with pollen/dust and cow milk allergy with the use of a multiplex Cytometric Bead Array (CBA) test. Twenty children with allergy: 11 with pollen/dust and 9 with cow milk allergy were enrolled into the study. For the analysis of INFγ, IL-2, IL-4, IL-6, IL-10, IL-17A, and TNFα levels, a flow cytometric test was used. TGFβ concentration was measured using an ELISA test. Concentrations of almost all of the detected cytokines in the patient’s plasma were below 1 pg/ml, with no significant difference between the groups examined. The level of IL-17A was higher in pollen/dust allergy group (median 12.73 pg/ml; 25th percentile-Q1/75th percentile-Q3; 9.28/15.83) in comparison to cow milk allergy group (median 7.86 pg/ml – Q1/Q3; 7.86/14.37). Children with cow milk allergy had a slightly higher concentration of TGFβ than children with pollen/dust allergy (median 28.7 ng/ml – Q1/Q3; 19.77/33.00 vs. 17.84 ng/ml – Q1/Q3; 11.21/23.75, respectively); the difference did not reach statistical significance. We conclude that plasma concentration of Th1, Th2, and Th17 regulatory cytokines are relatively low in allergic children. The CBA test, which could be useful in pediatric practice as it requires a relatively small plasma sample volume, is not sensitive enough to measure low cytokine levels, since its detection limit and the standard curve are not appropriate for the concentrations of these mediators expected in clinical samples.

Asthma can be effectively treated with sublingual immunotherapy. The influence of -sublingual immunotherapy on the function of granulocytes in asthmatic patients is largely unknown. Mac-1 integrin is a transmembrane protein containing α (CD11b) and β (CD18) chains. High expression of the complex is found on the surface of neutrophils, NK cells, and macrophages. CD11b/CD18 may bind to CD23, ICAM-1, ICAM-2, and ICAM-4. It plays a crucial role in diapedesis of neutrophils. The aim of the present study was to assess Mac-1 expression on neutrophils from asthmatic children before and after sublingual immunotherapy. Twenty five children aged of 8.1 ± 3.1 suffering from atopic asthma and allergic rhinitis, shortlisted for specific immunotherapy, served as the study group. Fifteen healthy individuals, aged 9.8 ± 3.4, served as a control group. The assessment of CD11b and CD18 expression on cells from peripheral blood was performed with a flow cytometer. The tests were performed before and after 12 months of sublingual immunotherapy. In the asthmatic children, 98.08 (90.79-99.12)% of Mac-1 positive neutrophils were detected. The group was divided into two subgroups: of more than 98% and less than 95% of neutrophils with CD11b/CD18 expression in the sample. After immunotherapy, the percentage of Mac-1 positive granulocytes increased to 99.60 (99.29-99.68)%, p = 0.01. In the control group, 90.56 (87.08-88.86)% granulocytes were Mac-1 positive, p = 0.002. We conclude that sublingual immunotherapy strongly influences the function of the immunological system, including Mac-1 expression on neutrophils.

Asthma can be effectively treated with sublingual immunotherapy. The influence of ­sublingual immunotherapy on the function of granulocytes in asthmatic patients is largely unknown. Mac-1 integrin is a transmembrane protein containing α (CD11b) and β (CD18) chains. High expression of the complex is found on the surface of neutrophils, NK cells, and macrophages. CD11b/CD18 may bind to CD23, ICAM-1, ICAM-2, and ICAM-4. It plays a crucial role in diapedesis of neutrophils. The aim of the present study was to assess Mac-1 expression on neutrophils from asthmatic children before and after sublingual immunotherapy. Twenty five children aged of 8.1 ± 3.1 suffering from atopic asthma and allergic rhinitis, shortlisted for specific immunotherapy, served as the study group. Fifteen healthy individuals, aged 9.8 ± 3.4, served as a control group. The assessment of CD11b and CD18 expression on cells from peripheral blood was performed with a flow cytometer. The tests were performed before and after 12 months of sublingual immunotherapy. In the asthmatic children, 98.08 (90.79–99.12)% of Mac-1 positive neutrophils were detected. The group was divided into two subgroups: of more than 98% and less than 95% of neutrophils with CD11b/CD18 expression in the sample. After immunotherapy, the percentage of Mac-1 positive granulocytes increased to 99.60 (99.29–99.68)%, p = 0.01. In the control group, 90.56 (87.08–88.86)% granulocytes were Mac-1 positive, p = 0.002. We conclude that sublingual immunotherapy strongly influences the function of the immunological system, including Mac-1 expression on neutrophils.

Atopic allergy is among the immune tolerance-related disorders resulting from a failure of the regulatory network. Regulatory T cells (Tregs) play a leading role in the development of homeostasis in the immune system. The aim of this study was to determine the role of Tregs in the pathogenesis of atopic diseases in children by exploring the relationship between Treg frequency, activation markers and the clinical manifestations of the disease.Twenty allergic and 50 healthy children were enrolled to the study. Peripheral blood mononuclear cells were stained with monoclonal antibodies (anti-CD25-CD4-CD127-FoxP3-CD69-CD71) and evaluated using flow cytometry. Tregs were identified as CD4+CD25(+/high)FoxP3+CD127- T cells.The percentage of Tregs in allergic patients (2.3%) was significantly decreased in comparison to healthy controls (4.6%, p = 0.003). The frequency of Tregs in patients with symptoms of atopic dermatitis and/or food allergy (1.7%) was significantly lower than in patients without these symptoms (2.9%, p = 0.04). A significant correlation between the percentage of Tregs and the sIgE serum concentration was observed (p = 0.037). Relative fluorescence intensities of FoxP3 expression in allergic patients were higher than in healthy controls (p = 0.00004). The frequency of CD4+CD25(high)CD127-CD71+ cells did not differ between the groups.Tregs display substantial deficiencies in atopic children, especially in children with multiorgan involvement, compared to patients with single organ manifestations. Additionally, there is an association between Tregs and the sIgE serum concentration. Better identification and characterization of Tregs in allergy is needed as they limit responses to foreign antigens, thereby minimizing T cell-mediated immunopathology in allergic diseases.

Background The resistance of T lymphocytes to Fas-mediated apoptosis is an important feature of atopic asthma. The only effective causative treatment of atopic diseases is immunotherapy. Clinical efficacy of sublingual immunotherapy (SLIT) has been already proven, but there is still limited number of studies on its influence on lymphocytes function. Objectives The aim of the study was to evaluate whether SLIT could restore the sensitivity of asthmatic T cells to undergo Fas-mediated apoptosis. Materials and methods Peripheral blood was collected from 12 patients aged 8 ± 2 years suffering from atopic asthma and undergoing sublingual specific immunotherapy. To evaluate sensitivity to Fas-mediated apoptosis, the blood was transmitted to sterile tubes and mixed with purified monoclonal antibody anti-CD95. After incubation, leukocytes were stained with Annexin V, propidium iodide, and monoclonal antibody against CD2 conjugated with phycoerythrincyanin 5.1, and then analyzed with flow cytometry. The procedure was repeated for each patient after 12 months of SLIT. Results Stimulation with anti-CD95 of T lymphocytes from patients with atopic asthma before treatment increased the number of early apoptotic cells (from 19.5 ± 16.7% before stimulation to 26.6 ± 16.7% Annexin V positive cells after stimulation). After one year of SLIT anti-CD95 still caused an increase of the early apoptotic cells ratio in the lymphocyte population (from 12.4 ± 7.4% before stimulation to 24.7 ± 15.4% Annexin V positive T cells after CD95 stimulation). Although an increasing trend could be observed, differences between the analyzed groups were not statistically significant. Conclusions A year of SLIT does not change the sensitivity of T lymphocytes from peripheral blood of children suffering from atopic asthma to Fas-mediated apoptosis.

IL-4 receptor (IL-4R) overexpression on immunoregulatory/effector cells was found in allergic patients. However, its role in allergy development remains unclear. The aim of this study was to assess the correlation between IL-4R expression and allergy development within the first year of life.IL-4R expression on monocytes and Th lymphocytes of 43 newborns was analyzed using flow cytometry. Plasma levels of IL-4, -12 and IFN-gamma were also measured using ELISA. The same parameters were assessed one year later. Furthermore, clinical evaluation was performed every three months for one year.Mean IL-4R expression on monocytes and Th lymphocytes did not differ at birth. After one year it increased on Th-lymphocytes and decreased on monocytes. However, among 10 children with severe atopy during the observation period, 8 displayed IL-4R above the mean value for the group on both monocytes and Th cells at birth as well as one year later. No correlation was found between IL-4 or IFN-gamma and IL-4R expression at birth. After one year, significant IL-4 increases and IFN-gamma decreases were observed which correlated with IL-4R expression. IL-4R expression on the newborns' monocytes correlated negatively with IL-12 plasma level; however, it was statistically significant only in the children developing allergy. Moreover, only in these patients was a significant decrease in IL-12 found after one year.IL-4R-dependent over-signaling in newborns' monocytes and Th lymphocytes could contribute to Th1/Th2 imbalance. IL-4R overexpression on newborns' monocytes and lymphocytes could be an early risk marker of allergy development.

Allergen-specific immunotherapy (AIT) is an effective method of allergy treatment. Reduction of allergy symptoms followed by AIT presumably depends on modification of cytokine production, especially of IFN-gamma and/or IL-4. Previously, we found that lower IL-12 production by allergic monocytes than in healthy control, could depend on higher IL-4 receptor alpha-chain (CD124) expression. Since IFN-gamma production is stimulated by IL-12, which in turn is down-regulated by IL-4, the aim of study was to analyze the influence of AIT on the network of these cytokines. Moreover, we estimated the possible role of CD124 in that process. Patients (n=16) with grass pollen allergy were subjected to AIT with Allergovit for 5 to 6 weeks. The clinical examination and blood analyses were performed before and after AIT. Clinical improvement in course of AIT was rather weak, however, we found significant increase of mean IFN-gamma production by PBMC. Although the increase of mean IL-12 production by monocytes following AIT was non-significant, we observed statistically significant decrease of monocyte sensitivity to IL-4 suppressed IL-12 production. Flow cytometry analysis revealed significant decrease of CD124 expression on monocytes after AIT. Despite unchanged mean IL-4 level the decrease of CD124 expression after AIT could explain the decreased monocyte sensitivity to IL-4 and its role in further IL-12 and IFN-gamma tuning.

The airway remodeling in asthma is associated with increased amount of matrix metalloproteinase (MMP)-9. High levels of MMP-9 were found in mucosal biopsies, sputum and in exhaled breath condensates (EBC) of asthma patients. However, there are no data concerning real in vivo activity. Inhaled corticosteroids are effective in asthma control, but it is unclear, whether they only attenuate inflammation, or also protect against progressive remodeling of respiratory tract. Therefore, the aim of the study was to assess the amount and activity of MMP-9 in context of pro-inflammatory cytokines (IL-6, IL-8 and tumor necrosis factor, TNF), measured in EBC of asthma-suffering children, treated with inhaled steroids. The study involved 27 children with asthma, continuously treated with inhaled fluticasone propionate, and 22 healthy controls. In addition to routine clinical screening, the selected cytokines in EBC were analyzed using Ultrasensitive ELISA, whereas activity of MMP-9 was assessed using a novel immunozymography method. Despite chronic treatment with inhaled steroids mean MMP-9/EBC activity in asthma group was significantly higher than in healthy controls. Moreover, high MMP-9/EBC in asthma-suffering children significantly correlated with IgE serum levels. The IL-6 and IL-8 concentration was below the detection limit in all EBC samples. TNF/EBC levels were similar in both, asthma and healthy children. We hypothesize that MMP-9 hyperactivity in asthma may be closely related to high IgE serum levels. Our results suggest that inhaled steroids may be ineffective to prevent asthma-associated airway remodeling. Finally, we emphasize the necessity of further research focused on MMP-9 inhibition in asthma treatment.