Evaluacija kriterijuma za dijagnozu atopijskog dermatitisa i detekcija alergen specifičnih IgE antitela kod pasa alergičnih na polen biljke Ambrosia artemisiifolia

Common ragweed (Ambrosia atremisiifolia) is one of the most frequent causes of pollen-induced allergic reactions both in humans and dogs. It has not been defined yet, what is the major allergen(s) to which most dogs allergic to ragweed show a positive result on intradermal skin test (IDST). In the present study sensitization to Ambrosia artemisiifolia pollen allergens in dogs with atopic dermatitis was examined with both in vivo and in vitro tests, including IDST and serum allergen specific IgE test. Detection of specific-IgE antibodies against ragweed allergens by immunoblotting in the sera of allergic dogs was optimized, as well. Dogs that were positive, as judged by IDST reactions to ragweed pollen allergens, also had alergen specific IgE antibodies in their sera. Results indicate that major allergens of A. artemisifolia pollen in dogs are Amb a 1 and Amb a 2. Further characterization of ragweed allergens is needed before they could potentially be used in intradermal testing or allergen immunotherapy in affected dogs. Also, we evaluated new Favrots diagnostic criteria for canine atopic dermatitis in dogs allergic to Ambrosia atremisiifolia pollen. It might be concluded that proposed criteria are of great assistance for seting up suspected diagnosis of canine atopic dermatitis, after ruling out other pruritic dermatoses.Kratka ambrozija (Ambrosia artemisiifolia) je jedan od najčešćih uzročnika alergijskih reakcija izazvanih polenom kod ljudi i pasa. Još uvek nije definisano koji je glavni alergen (i), na koji, većina pasa alergičnih na polen ambrozije, ispoljava pozitivnu reakciju na intradermalnom testu. U ovoj studiji je ispitana senzibilizacija na polen ove biljke kod pasa sa simptomima atopijskog dermatitisa in vivo i in vitro testovima, uključujući intradermalni test i dokazivanje prisustva alergen specifičnih IgE antitela u serumu. Optimizovani su uslovi za detekciju IgE specifičnih antitela iz seruma pasa alergičnih na polen ambrozije imunoblot tehnikom. Psi koji su imali pozitivnu reakciju na polen ove biljke na intradermalnom testu, takođe su imali specifična IgE antitela u serumu. Dobijeni rezultati ukazuju da su glavni alergeni Ambrosia artemisiifolia kod pasa Amb a 1 i Amb a 2. Neophodna je dalja karakterizacija alergena ambrozije kako bi se oni mogli primeniti pri rutinskom intradermalnom testiranju ili u alergen specifičnoj imunoterapiji obolelih pasa. Takođe je razmatrana i validnost Favrotovih dijagnostičkih kriterijuma kod pasa alergičnih na polen ambrozije. Može se zaključiti da su predloženi kriterijumi od velike pomoći u postavljanju suspektne dijagnoze atopijskog dermatitisa pasa, nakon isključenja drugih pruritičnih dermatoza

Rana alergijska reakcija na metilprednizolon sa tolerancijom drugih kortikosteroida

Introduction. In spite of the wide usage of corticosteroids for the treatment of a plethora of diseases, sometimes they can induce immediate hypersensitivity reactions, which are however uncommon. Case Outline. We report a case of immediate allergic reaction induced by intravenous methylprednisolone given before operation for surgical repair of an arm contracture as a sequel of burns, which the child had tolerated a month before. Six weeks later the patient repeated the anaphylactic reaction during skin testing to methylprednisolone. In addition, basophile activation test with methylprednisolone (BAT) was positive. Conclusion. This case report describes a patient who experienced intraoperative anaphylaxis and anaphylactic reaction induced by skin testing. This is the first report on induction of both anaphylactic reactions by methylprednisolone in the same child. Clinical findings, positive BAT and positive skin tests with methylprednisolone imply that the child developed type-I hypersensitivity. The lack of cross-reactivity with other corticosteroids emphasizes that the reactions were caused by the steroid molecule.Uvod. Uprkos širokoj primeni kortikosteroida u lečenju od različitih bolesti, oni ponekad mogu izazvati ranu alergijsku reakciju. Prikaz bolesnika. Kod dvanaestogodišnjeg dečaka došlo je do rane alergijske reakcije izazvane intravenskom primenom metilprednizolona neposredno pre hirurške intervencije, tačnije, korekcije kontrakture šake koja se javila kao komplikacija opekotine. Mesec dana pre pojave alergijske reakcije dete je primalo metilprednizolon i dobro ga podnosilo. Šest nedelja posle operacije ponovo se javila anafilaktička reakcija tokom kožnog testiranja metilprednizolonom. Primenjen je i test aktivacije bazofila (BAT) ovim lekom, čiji je nalaz bio pozitivan. Zaključak. Ovo je prvi prikaz dve vrste anafilaktičke reakcije izazvane metilprednizolonom kod iste osobe. Klinička slika, pozitivni nalaz BAT i pozitivne kožne probe na metilprednizolon pokazuju da se kod deteta razvio prvi tip hipersenzitivne reakcije. Nedostatak unakrsne reaktivnosti s ostalim kortikosteroidima ukazuje na to da je alergijska reakcija izazvana steroidnim molekulom

Evaluation of the immunomodulatory potential of chimera Bv1a-BLwt and its mutants on the co-culture model system

Allergen immunotherapy (AIT) is currently the only disease-modifying treatment forallergies. Pre-clinical models for the evaluation of novel therapeutics are crucial forensuring their efficacy and safety. While cell culture models are cost-effective andefficient, they cannot fully replicate the cellular interactions in vivo. Therefore, it isessential to use more sophisticated model systems, such as co-cultures, to assess thepotential of new therapeutics more accurately. Immunomodulatory protein banana lectin(BLwt) is an attractive candidate for adjuvant in AIT. Its mutant BLH84T was developed toreduce its potential mitogenicity. The aim of this study was the development of the coculture model system for testing the immunomodulatory effect of chimeras composed ofthe major birch pollen allergen (Bv1a) and BLwt (Bv1a-BLwt, Cwt), the hypoallergenicisoform of Bv1a (Bv1l) and BLH84T (Bv1l-BLH84T, C1 and BLH84T-Bv1l, C2). Chimericstructures were designed in silico, fully minimized, and relaxed without van der Waalsatomic clashes. Afterward, proteins were successfully expressed in Escherichia coli andpurified by IMAC yielding around 0.4 mg per 1L of expression medium. The IgE bindingcapacity was assessed using ELISA inhibition with birch pollen allergic patients’ sera.Caco-2 intestinal epithelial cells and THP-1 differentiated macrophages were used for theco-culture model system development. After protein application on the apical side of theco-culture, the integrity of the epithelial monolayer was not disturbed. Theimmunomodulatory potential of antigens was tested by measuring the gene expressionlevels for pro- and anti-inflammatory cytokines in both cell lines from co-culture. Theobtained results indicate that the best anti-inflammatory response was favored aftertreatment with Cwt. Additionally, to further confirm the immunomodulatory effect of therecombinant chimeras, PBMCs obtained from individuals allergic to birch pollen wereemployed and treated with recombinant proteins. Only after treatment with Cwt, PBMCssecreted the anti-inflammatory cytokine IL-10. Obtained results suggest that Cwt chimeracould have a therapeutic effect in AIT in birch pollen allergy

Evaluation of the immunomodulatory potential of chimera Bv1a-BLwt and its mutants on the co-culture model system

Allergen immunotherapy (AIT) is currently the only disease-modifying treatment for allergies. Pre-clinical models for the evaluation of novel therapeutics are crucial for ensuring their efficacy and safety. While cell culture models are cost-effective and efficient, they cannot fully replicate the cellular interactions in vivo. Therefore, it is essential to use more sophisticated model systems, such as co-cultures, to assess the potential of new therapeutics more accurately. Immunomodulatory protein banana lectin (BLwt) is an attractive candidate for adjuvant in AIT. Its mutant BLH84T was developed to reduce its potential mitogenicity. The aim of this study was the development of the coculture model system for testing the immunomodulatory effect of chimeras composed of the major birch pollen allergen (Bv1a) and BLwt (Bv1a-BLwt, Cwt), the hypoallergenic isoform of Bv1a (Bv1l) and BLH84T (Bv1l-BLH84T, C1 and BLH84T-Bv1l, C2). Chimeric structures were designed in silico, fully minimized, and relaxed without van der Waals atomic clashes. Afterward, proteins were successfully expressed in Escherichia coli and purified by IMAC yielding around 0.4 mg per 1L of expression medium. The IgE binding capacity was assessed using ELISA inhibition with birch pollen allergic patients’ sera. Caco-2 intestinal epithelial cells and THP-1 differentiated macrophages were used for the co-culture model system development. After protein application on the apical side of the co-culture, the integrity of the epithelial monolayer was not disturbed. The immunomodulatory potential of antigens was tested by measuring the gene expression levels for pro- and anti-inflammatory cytokines in both cell lines from co-culture. The obtained results indicate that the best anti-inflammatory response was favored after treatment with Cwt. Additionally, to further confirm the immunomodulatory effect of the recombinant chimeras, PBMCs obtained from individuals allergic to birch pollen were employed and treated with recombinant proteins. Only after treatment with Cwt, PBMCs secreted the anti-inflammatory cytokine IL-10. Obtained results suggest that Cwt chimera could have a therapeutic effect in AIT in birch pollen allergy

Quantifying the Effects of Drought Using the Crop Moisture Stress as an Indicator of Maize and Sunflower Yield Reduction in Serbia

The drought in Serbia in the summer of 2017 heavily affected agricultural production, decreasing yields of maize, sunflower, soybean, and sugar beet. Monitoring moisture levels in crops can provide timely information about potential risk within a growing season, thus helping to create an early warning system for various stakeholders. The purpose of this study was to quantify the level of moisture stress in crops during summer and the consequences that it can have on yields. For that, maize and sunflower yield data provided by an agricultural company were used at specific parcels in the Backa region of Vojvodina province (Serbia) for 2017, 2018, 2019, and 2020. The crop moisture level was estimated at each parcel by calculating the normalized difference moisture index (NDMI) from Sentinel-2 data during the summer months (June–July–August). Based on the average NDMI value in July, the new crop moisture stress (CMS) index was introduced. The results showed that the CMS values at a specific parcel could be used for within-season estimation of maize and sunflower yield and the assessment of drought effects. The CMS index was tested for the current growing season of 2022 as an early warning system for yield reduction, demonstrating the potential to be included in a platform for digital agriculture, such as AgroSens, which is operational in Serbia

rBet v 1a-BanLec_wt induce upregulation of IL-10 and IFN-γ gene expression in Caco-2/THP-1 co-culture and secretion of IL-10 and IFN-γ/IL-4 levels in PBMCs of birch pollen allergic donors

Novel allergen immunotherapy (AIT) approaches necessitate the use of more effective and safe therapeutics, which can be accomplished by employing novel adjuvants for improved innate immune cell activation, as well as hypoallergenic allergen forms. In this study, we investigate the immunomodulatory effects of a chimera rBet v 1a-BanLecwt (rBv1a-BLwt; Cwt) composed of the major birch pollen allergen Bet v 1a and banana lectin (BanLecwt; BLwt) and two novel chimeras, rBv1l-BLH84T (rBet v 1l-BanLecH84T; C1) and rBLH84T-Bv1l (rBanLecH84T-Bet v 1l; C2), both composed of BLH84T and hypoallergenic birch pollen allergen Bv1l in the co-culture model Caco-2/THP-1, and PBMCs from donors with birch pollen allergy. The chimeric molecules rBv1l-BLH84T (C1) and rBLH84T-Bv1l (C2) were created in silico and then produced in E. coli using recombinant DNA technology. Real-time PCR analysis of gene expression following compound treatment in the co-culture model revealed that all three chimeras have the potential to induce the anti-inflammatory cytokine IL-10 gene expression in Caco-2 cells and IFN-γ gene expression in THP-1 cells. Sandwich ELISA revealed that Cwt increased IL-10 secretion and IFN-/IL-4 levels in PBMCs from birch pollen allergic donors, whereas C1 and C2 were less effective. The findings suggest that Cwt should be analyzed further due to its potential benefit in AIT