The influence of sublingual immunotherapy on several parameters of immunological response in children suffering from atopic asthma and allergic rhinitis depending on asthma features

Wstęp: Kliniczna skuteczność immunoterapii podjęzykowej została już gruntowanie potwierdzona i udokumentowana. Badano również jej wpływ na układ odpornościowy chorych na astmę oskrzelową. J ednak do tej pory nie wyjaśniono zależności pomiędzy występowaniem alergii wieloważnej, współwystępowaniem innych chorób alergicznych i stopniem terapii astmy a wpływem prowadzonej alergenowo-swoistej immunoterapii podjęzykowej. Celem niniejszej pracy była ocena wpływu rocznej immunoterapii podjęzykowej na wybrane parametry odpowiedzi immunologicznej w zależności od cech osobniczych i klinicznych cech choroby u pacjentów chorych na astmę i alergiczny nieżyt nosa.Materiał i metody: Do badania zakwalifikowano 25 dzieci w wieku 8,1 ± 3,1 roku (5–15 lat), 21 chłopców i 4 dziewczynki, chorych na astmę oskrzelową i alergiczny nieżyt nosa z alergiami poliwalentnymi sezonowymi i niesezonowymi, które były zakwalifikowane do alergenowo-swoistej immunoterapii podjęzykowej. Odsetek limfocytów pomocniczych Th1 i Th2, ekspresję antygenu Bcl-2 w limfocytach T oraz aktywację bazofilów pod wpływem swoistych alergenów (zawiesiny antygenów roztoczy kurzu domowego i/lub pyłków traw do testów skórnych) badano metodą cytometrii przepływowej. Z ależności pomiędzy badanymi parametrami oceniano dokładnym testem Fishera.Wyniki: Nie zaobserwowano istotnych zależności pomiędzy wpływem immunoterapii podjęzykowej na układ odpornościowy pacjentów a wiekiem, występowaniem alergii wieloważnej, stopniem terapii astmy i współwystępowaniem innych chorób alergicznych. Wzrost odsetka limfocytów pomocniczych Th1 pod wpływem immunoterapii podjęzykowej obserwowano nieco częściej u dzieci uczulonych na więcej niż 3 alergeny niż u pozostałych.Wnioski: Na podstawie badań przeprowadzonych przez autorów pracy nie można wskazać żadnej grupy pacjentów, u których można się spodziewać silniejszej mobilizacji układu odpornościowego jako odpowiedzi na prowadzoną immunoterapię podjęzykową u dzieci chorych na astmę i alergiczny nieżyt nosa. Wzrost odsetka limfocytów pomocniczych Th1 może występować częściej u dzieci z alergią poliwalentną.Introduction: The clinical efficacy of sublingual immunotherapy (SLIT) has already been proven and is known to be high. Its influence on the immunological system of patients suffering from bronchial asthma was also examined. However, it is still unclear how the polysensitisation, coexistence of other atopic disease and asthma treatment step influence the response to treatment with specific immunotherapy. Herein we evaluate the impact of one-year SLIT on selected markers of immunological response depending on different individual and clinical factors of children suffering from atopic asthma and allergic rhinitis.Material and methods: Twenty-five patients aged 8.1 ± 3.1 years (range 5–15 years), 21 boys and 4 girls, suffering from asthma and allergic rhinitis with polysensitisation to seasonal and non-seasonal allergens, shortlisted for SLIT, were included in the study. Th1 cell and Th2 cell percentages, Bcl-2 expression in T cells, and basophil activation after allergen challenge (house dust mite and/or grass pollen antigen in solution used for skin prick tests) in peripheral blood were measured using flow cytometry. The association between clinical features of asthma and the influence of SLIT on immunological parameters was evaluated with exact Fisher test.Results: No association between the influence of one-year sublingual immunotherapy on immunological system and patients’ age, polysensitisation, asthma treatment step, or coexistence of any other atopic diseases was observed. However, an increase of the Th1 percentage in children sensitised against more than three allergens was found more often (at the limit of statistical significance) than in the group of children sensitised against three or less allergens.Conclusions: Based on our results, we cannot point to any subgroup isolated in the study, in which the response of the immunological system to sublingual immunotherapy is more satisfactory than any other. Nevertheless, the increase of Th1 cells may be more specific for polysensitised children

FoxP3 Tregs Response to Sublingual Allergen Specific Immunotherapy in Children Depends on the Manifestation of Allergy

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

Lymphocytes sensitivity to Fas stimulation in healthy and asthmatic children.

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

The usefulness of CD203c expression measurement on basophils after activation with grass pollen and Dermatophagoides pteronyssinus antigens. Preliminary study

Wstęp: "Złotym standardem" w diagnostyce chorób atopowych są testy skórne i oznaczanie poziomu swoistego IgE. Obecnie możliwa jest również cytometryczna ocena aktywacji bazofilów jako komórek efektorowych w reakcjach atopowych. Celem pracy była ocena przydatności testu in vitro wykorzystującego ekspresję CD203c jako markera aktywacji bazofilów w diagnostyce atopii. Materiał i metody: Przebadano 22 osoby, w tym 13 chorych na astmę atopową, potwierdzoną wynikami punktowych testów skórnych, zakwalifikowanych do immunoterapii swoistej, oraz 9 osób zdrowych, z ujemnymi wynikami testów skórnych. Ekspresję antygenu CD203c oceniano z wykorzystaniem cytometru przepływowego Cytomics FC500 (Beckman Coulter) za pomocą testu Allergenicity Kit (Beckman Coulter). Wyniki: Za wynik dodatni uznano aktywację bazofilów na poziomie > 15% w stosunku do kontroli negatywnej. U wszystkich chorych na astmę atopową wykazano podwyższony poziom aktywacji bazofilów powyżej poziomu odcięcia. Średni poziom aktywacji bazofilów pod wpływem antygenów pyłków traw dla osób uczulonych wyniósł 50,95 &#177; 15,7% (mediana 49,7%, zakres 1,91-72,42%) w stosunku do 1,91% (0,00-7,96%) dla osób zdrowych (p = 0,002). Z kolei poziom aktywacji bazofilów pod wpływem antygenów D. pteronyssinus wyniósł 40,6 &#177; 25,2% u osób uczulonych, zaś u osób nieuczulonych - 2,51 &#177; 1,96% (p = 0,0003). Bazofile 21 badanych osób odpowiadały na stymulator kontrolny (anty-IgE) (44,1 &#177; 18,9%). W żadnej z badanych próbek nie uzyskano pobudzenia pod wpływem kontroli negatywnej (2,03 &#177; 1,19%, p < 0,0001). Czułość i swoistość zastosowanego testu wyniosły odpowiednio 92,3 i 100%. Wnioski: Test z wykorzystaniem detekcji antygenu CD203c może być bardzo użyteczny w diagnostyce alergii i - być może - w monitorowaniu immunoterapii swoistej.Introduction: 'Gold standard' in the diagnosis of atopic disease are skin prick tests and specific IgE evaluation. Well-established in vitro tests, such as the histamine release test, the leukotriens release test and the flow cytometric basophil activation test can be very helpful in diagnostics, especially when the skin prick test is contraindicated. The aim of this study was to evaluate the usefulness of antigen CD203c expression, as a marker of basophil activation by grass pollen or D. pteronyssinus antigens. Material and methods: Peripheral blood from 13 allergic patients and nine healthy volunteers was analysed. Basophils activation was measured by the breakdown of antigen CD203c expression with Allergenicity Kit (Beckman Coulter), using Cytomics FC 500 flow cytometer (Beckman Coulter). Results: The sensitivity was 92,3% and specificity of test was 100%. 50.95 &#177; 15.7% of basophils (median 49.7%, 1.91-72.42%) were activated after grass pollen stimulation in atopic patients sensitised to this allergen, in comparison to 1.91% (0.00-7.96%) in control patients (p = 0.002). The percentage of activated basophils after D. pteronyssinus antigens stimulation was 40.6 &#177; 25.2% in allergic patients, compared to only 2.51 &#177; 1 96% of basophils from non-atopic controls (p = 0.0003). Basophils from 21 patients responded after anti-IgE stimulation (44.1 &#177; 18.9%), and none of the analysed samples was activated after PBS stimulation (2.03 &#177; 1.19%, p < 0.0001). Conclusions: These results demonstrate that basophil activation test based on antigen CD203c expression is very accurate in the diagnosis of atopic diseases

Serum 25(OH)D status and lipid profile in children with newly diagnosed asthma

Background. The problem of the influence of hyperlipidemia on asthma was addressed several years ago. Systematic review and meta-analysis performed in the pediatric population on the association between vitamin D status and lipid profile components revealed discordant results and indicated that higher serum 25(OH)D is related to a more favorable lipid profile. Objective. We aimed to elucidate whether there was an association between vitamin D status and lipid profile components and apolipoprotein B in a sample of children with newly diagnosed atopic asthma. Methods. The study included 150 children aged 2–12 years. Atopic asthma was diagnosed in 110 children; 40 children constituted a reference group. Fasting blood was collected to measure 25(OH)D total, lipid profile and apolipoprotein B concentrations. Results. Children with asthma had significantly lower 25(OH)D (p &lt; 0.0001) but similar lipid and apolipoprotein B concentrations. The proportions of hypercholesterolemia, hypertriglyceridemia and increased apoB concentrations were similar in both groups. HDL-C concentrations in asthmatic 25(OH)D-deficient children were higher compared with the children with sufficient levels (p = 0.05). ApoB concentration was lower in 25(OH)D-deficient compared with vitamin D sufficient asthmatics (p = 0.0008). Correlations between 25(OH)D concentration and lipids and apoB in asthmatics revealed gender differences. An inverse relationships between vitamin D and total cholesterol and HDL-C (R= –0.39, p &lt; 0.05; R= –0.475, p &lt; 0.001) were found in girls. In boys vitamin D correlated with LDL-C and apoB (R = 0.376, R = 0.498; p &lt; 0.001). Conclusion. In children with asthma lower 25(OH)D had more favorable gender-dependent effect on the lipid profile. The association of serum 25(OH)D and lipid levels in children with asthma remains for further studies.

Lymphocytes sensitivity to Fas stimulation in healthy and asthmatic children.

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

25-hydroxyvitamin D insufficiency in children with newly diagnosed asthma

Background. 25-hydroxyvitamin D [25(OH)D] deficiency seems to be related to the development of asthma. Any evaluation of the relationship between asthma and 25(OH)D deficiency must consider the association between increased airway responsiveness, eosinophil counts and serum immunoglobulin E (IgE), and 25(OH)D as a potential player in airway remodelling. Objective. We assessed the association of 25(OH)D with markers of atopy and eosinophilic inflammation in children with newly diagnosed asthma. Methods. The study included 165 children aged 2–12 years. The diagnosis of asthma was performed by an experienced paediatric pulmonologist. Allergic asthma was diagnosed in 106 children, and non-allergic asthma in ten; in 49 children, asthma was excluded. Fasting blood was collected for cell counts, and serum was obtained to measure lipids, C-reactive protein (hsCRP), 25(OH)D and IgE. Results. Children with asthma had significantly lower 25(OH)D (p &lt; 0.001). Both groups had similar lipid values. Elevated total IgE concentration and eosinophil counts were found in asthmatics; neutrophils were similar in asthmatic and reference groups. There was a strong tendency to higher eosinophil counts in 25(OH)D-deficient children (&lt; 20 ng/mL) with atopic asthma (p &lt; 0.08). Conclusion. In children with asthma, 25(OH)D insufficiency/deficiency is associated with higher eosinophil counts and IgE. 25(OH)D monitoring is important in the prevention and management of children with asthm

FoxP3 Tregs Response to Sublingual Allergen Specific Immunotherapy in Children Depends on the Manifestation of Allergy

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

FoxP3 Tregs Response to Sublingual Allergen Specific Immunotherapy in Children Depends on the Manifestation of Allergy

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