Alternaria alternata is a common fungus strongly related with severe allergic asthma, with 80% of affected individuals being sensitized solely to its major allergen Alt a 1. Here, we assessed the function of Alt a 1 as an innate defense protein binding to micronutrients, such as iron-quercetin complexes (FeQ2), and its impact on antigen presentation in vitro. Binding of Alt a 1 to FeQ2 was determined in docking calculations. Recombinant Alt a 1 was generated, and binding ability, as well as secondary and quaternary structure, assessed by UV-VIS, CD, and DLS spectroscopy. Proteolytic functions were determined by casein and gelatine zymography. Uptake of empty apo- or ligand-filled holoAlt a 1 were assessed in human monocytic THP1 cells under the presence of dynamin and clathrin-inhibitors, activation of the Arylhydrocarbon receptor (AhR) using the human reporter cellline AZ-AHR. Human PBMCs were stimulated and assessed for phenotypic changes in monocytes by flow cytometry. Alt a 1 bound strongly to FeQ2 as a tetramer with calculated Kd values reaching pico-molar levels and surpassing affinities to quercetin alone by a factor of 5000 for the tetramer. apoAlt a 1 but not holoAlta 1 showed low enzymatic activity against casein as a hexamer and gelatin as a trimer. Uptake of apo- and holo-Alt a 1 occurred partly clathrin-dependent, with apoAlt a 1 decreasing labile iron in THP1 cells and holoAlt a 1 facilitating quercetin-dependent AhR activation. In human PBMCs uptake of holoAlt a 1 but not apoAlt a 1 significantly decreased the surface expression of the costimulatory CD86, but also of HLADR, thereby reducing effective antigen presentation. We show here for the first time that the presence of nutritional iron complexes, such as FeQ2, significantly alters the function of Alt a 1 and dampens the human immune response, thereby supporting the notion that Alt a 1 only becomes immunogenic under nutritional deprivation.

Bioresources Unit Center for Health and Bioresources AIT Austrian Institute of Technology GmbH 3430 Tulln Austria

Comparative Medicine The Interuniversity Messerli Research Institute 1210 Vienna Austria

Department of Biosciences University of Salzburg 5020 Salzburg Austria

Department of Cell Biology and Genetics Faculty of Science Palacky University 779 00 Olomouc Czech Republic

Institute of Pathophysiology and Allergy Research Center of Pathophysiology Infectiology and Immunology Medical University of Vienna 1090 Vienna Austria

See more in PubMed

Lindblom S.D., Wangeline A.L., Barillas J.R.V., Devilbiss B., Fakra S.C., Pilon-Smits E.A.H. Fungal Endophyte Alternaria tenuissima Can Affect Growth and Selenium Accumulation in Its Hyperaccumulator Host Astragalus bisulcatus. Front. Plant Sci. 2018;9:1213. doi: 10.3389/fpls.2018.01213. PubMed DOI PMC

DeMers M. Alternaria alternata as endophyte and pathogen. Microbiology. 2022;168 doi: 10.1099/mic.0.001153. PubMed DOI PMC

Kumari V.V., Banerjee P., Verma V.C., Sukumaran S., Chandran M.A.S., Gopinath K.A., Venkatesh G., Yadav S.K., Singh V.K., Awasthi N.K. Plant Nutrition: An Effective Way to Alleviate Abiotic Stress in Agricultural Crops. Int. J. Mol. Sci. 2022;23:8519. doi: 10.3390/ijms23158519. PubMed DOI PMC

Vicki A.M., Daniel J.W. Alternaria: A Sinonasal Pathogen of Immunocompromised Hosts. Clin. Infect. Dis. 1993;16:265–270. PubMed

Yu H. Studies on fungi of the normal skin. Hifuka kiyo. Acta Dermatol. 1965;60:126–174. PubMed

Pastor F.J., Guarro J. Alternaria infections: Laboratory diagnosis and relevant clinical features. Clin. Microbiol. Infect. 2008;14:734–746. doi: 10.1111/j.1469-0691.2008.02024.x. PubMed DOI

Nema H.V., Ahuja O.P., Bal A., Mohapatra L.N. Mycotic Flora of the Conjunctiva. Am. J. Ophthalmol. 1966;62:968–970. doi: 10.1016/0002-9394(66)91928-3. PubMed DOI

Lo Porto D., Cona A., Todaro F., De Carolis E., Cardinale F., Hafeez N., Di Martino G., Conaldi P.G., Sanguinetti M., Grossi P.A., et al. Phaeohyphomycosis in Solid Organ Transplant Recipients: A Case Series and Narrative Review of the Literature. J. Fungi. 2023;9:283. doi: 10.3390/jof9030283. PubMed DOI PMC

Lee J.Y., Hyun M., Kim H.A., Ryu S.Y. Unusual Presentation of Subcutaneous Phaeohyphomycosis by Alternaria alternate. Ann. Dermatol. 2019;31:563–566. doi: 10.5021/ad.2019.31.5.563. PubMed DOI PMC

Hattab Z., Ben Lasfar N., Abid M., Bellazreg F., Fathallah A., Hachfi W., Letaief A. Alternaria alternata infection causing rhinosinusitis and orbital involvement in an immunocompetent patient. New Microbes New Infect. 2019;32:100561. doi: 10.1016/j.nmni.2019.100561. PubMed DOI PMC

Sanchez P., Velez-Del-Burgo A., Sunen E., Martinez J., Postigo I. Fungal Allergen and Mold Allergy Diagnosis: Role and Relevance of Alternaria alternata Alt a 1 Protein Family. J. Fungi. 2022;8:277. doi: 10.3390/jof8030277. PubMed DOI PMC

Kespohl S., Raulf M. Mold Sensitization in Asthmatic and Non-asthmatic Subjects Diagnosed with Extract-Based Versus Component-Based Allergens. Adv. Exp. Med. Biol. 2019;1153:79–89. doi: 10.1007/5584_2019_342. PubMed DOI

Chruszcz M., Chapman M.D., Osinski T., Solberg R., Demas M., Porebski P.J., Majorek K.A., Pomes A., Minor W. Alternaria alternata allergen Alt a 1: A unique beta-barrel protein dimer found exclusively in fungi. J. Allergy Clin. Immunol. 2012;130:241–247.e249. doi: 10.1016/j.jaci.2012.03.047. PubMed DOI PMC

Garrido-Arandia M., Silva-Navas J., Ramirez-Castillejo C., Cubells-Baeza N., Gomez-Casado C., Barber D., Pozo J.C., Melendi P.G., Pacios L.F., Diaz-Perales A. Characterisation of a flavonoid ligand of the fungal protein Alt a 1. Sci. Rep. 2016;6:33468. doi: 10.1038/srep33468. PubMed DOI PMC

Mechaly A.E., Ibanez de Opakua A., Asturia J., Viguera A.R. Crystal Structure of Alternaria alternata Major Allergen Alt a 1. [(accessed on 21 November 2021)]. Available online: https://www.wwpdb.org/pdb?id=pdb_00004aud.

Garrido-Arandia M., Bretones J., Gomez-Casado C., Cubells N., Diaz-Perales A., Pacios L.F. Computational study of pH-dependent oligomerization and ligand binding in Alt a 1, a highly allergenic protein with a unique fold. J. Comput. Aided Mol. Des. 2016;30:365–379. doi: 10.1007/s10822-016-9911-6. PubMed DOI

Regner A., Szepannek N., Wiederstein M., Fakhimahmadi A., Paciosis L.F., Blokhuis B.R., Redegeld F.A., Hofstetter G., Dvorak Z., Jensen-Jarolim E., et al. Binding to Iron Quercetin Complexes Increases the Antioxidant Capacity of the Major Birch Pollen Allergen Bet v 1 and Reduces Its Allergenicity. Antioxidants. 2022;12:42. doi: 10.3390/antiox12010042. PubMed DOI PMC

Afify S.M., Regner A., Pacios L.F., Blokhuis B.R., Jensen S.A., Redegeld F.A., Pali-Scholl I., Hufnagl K., Bianchini R., Guethoff S., et al. Micronutritional supplementation with a holoBLG-based FSMP (food for special medical purposes)-lozenge alleviates allergic symptoms in BALB/c mice: Imitating the protective farm effect. Clin. Exp. Allergy. 2021;52:426–441. doi: 10.1111/cea.14050. PubMed DOI

Afify S.M., Pali-Scholl I., Hufnagl K., Hofstetter G., El-Bassuoni M.A.-R., Roth-Walter F., Jensen-Jarolim E. Bovine Holo-Beta-Lactoglobulin Cross-Protects Against Pollen Allergies in an Innate Manner in BALB/c Mice: Potential Model for the Farm Effect. Front. Immunol. 2021;12:176. doi: 10.3389/fimmu.2021.611474. PubMed DOI PMC

Roth-Walter F. Iron-Deficiency in Atopic Diseases: Innate Immune Priming by Allergens and Siderophores. Front. Allergy. 2022;3:859922. doi: 10.3389/falgy.2022.859922. PubMed DOI PMC

Roth-Walter F., Afify S.M., Pacios L.F., Blokhuis B.R., Redegeld F., Regner A., Petje L.M., Fiocchi A., Untersmayr E., Dvorak Z., et al. Cow’s milk protein beta-lactoglobulin confers resilience against allergy by targeting complexed iron into immune cells. J. Allergy Clin. Immunol. 2021;147:321–334.e324. doi: 10.1016/j.jaci.2020.05.023. PubMed DOI

Roth-Walter F., Pacios L.F., Gomez-Casado C., Hofstetter G., Roth G.A., Singer J., Diaz-Perales A., Jensen-Jarolim E. The major cow milk allergen Bos d 5 manipulates T-helper cells depending on its load with siderophore-bound iron. PLoS ONE. 2014;9:e104803. doi: 10.1371/journal.pone.0104803. PubMed DOI PMC

Luscher A., Gasser V., Bumann D., Mislin G.L.A., Schalk I.J., Kohler T. Plant-Derived Catechols Are Substrates of TonB-Dependent Transporters and Sensitize Pseudomonas aeruginosa to Siderophore-Drug Conjugates. mBio. 2022;13:e0149822. doi: 10.1128/mbio.01498-22. PubMed DOI PMC

Raza A., Xu X., Xia L., Xia C., Tang J., Ouyang Z. Quercetin-Iron Complex: Synthesis, Characterization, Antioxidant, DNA Binding, DNA Cleavage, and Antibacterial Activity Studies. J. Fluoresc. 2016;26:2023–2031. doi: 10.1007/s10895-016-1896-y. PubMed DOI

Symonowicz M., Sykula-Zajac A., Lodyga-Chruscinska E., Rumora I., Straukas M. Evaluation of polyphenols and anthocyanins contents in black chockeberry—Photinia melanocarpa (Michx.) fruits extract. Acta Pol. Pharm. 2012;69:381–387. PubMed

El Hajji H., Nkhili E., Tomao V., Dangles O. Interactions of quercetin with iron and copper ions: Complexation and autoxidation. Free Radic. Res. 2006;40:303–320. doi: 10.1080/10715760500484351. PubMed DOI

Perron N.R., Brumaghim J.L. A review of the antioxidant mechanisms of polyphenol compounds related to iron binding. Cell Biochem. Biophys. 2009;53:75–100. doi: 10.1007/s12013-009-9043-x. PubMed DOI

Raymond K.N., Dertz E.A., Kim S.S. Enterobactin: An archetype for microbial iron transport. Proc. Natl. Acad. Sci. USA. 2003;100:3584–3588. doi: 10.1073/pnas.0630018100. PubMed DOI PMC

Xiao L., Luo G., Tang Y., Yao P. Quercetin and iron metabolism: What we know and what we need to know. Food Chem. Toxicol. 2018;114:190–203. doi: 10.1016/j.fct.2018.02.022. PubMed DOI

Wang X., Li Y., Han L., Li J., Liu C., Sun C. Role of Flavonoids in the Treatment of Iron Overload. Front. Cell Dev. Biol. 2021;9:685364. doi: 10.3389/fcell.2021.685364. PubMed DOI PMC

Yin M., Liu Y., Chen Y. Iron metabolism: An emerging therapeutic target underlying the anti-cancer effect of quercetin. Free Radic. Res. 2021;55:296–303. doi: 10.1080/10715762.2021.1898604. PubMed DOI

Lesjak M., Hoque R., Balesaria S., Skinner V., Debnam E.S., Srai S.K., Sharp P.A. Quercetin inhibits intestinal iron absorption and ferroportin transporter expression in vivo and in vitro. PLoS ONE. 2014;9:e102900. doi: 10.1371/journal.pone.0102900. PubMed DOI PMC

Bardestani A., Ebrahimpour S., Esmaeili A., Esmaeili A. Quercetin attenuates neurotoxicity induced by iron oxide nanoparticles. J. Nanobiotechnol. 2021;19:327. doi: 10.1186/s12951-021-01059-0. PubMed DOI PMC

Sajadi Hezaveh Z., Azarkeivan A., Janani L., Hosseini S., Shidfar F. The effect of quercetin on iron overload and inflammation in beta-thalassemia major patients: A double-blind randomized clinical trial. Complement. Ther. Med. 2019;46:24–28. doi: 10.1016/j.ctim.2019.02.017. PubMed DOI

Roth-Walter F., Gomez-Casado C., Jensen-Jarolim E., Diaz Perales A., Pacios L.F., Singer J. Method and Means for Diagnosing and Treating Allergy Using Lipocalin Levels. U.S. Patent No. 10,914,744. 2015 January 7;

Roth-Walter F., Gomez-Casado C., Pacios L.F., Mothes-Luksch N., Roth G.A., Singer J., Diaz-Perales A., Jensen-Jarolim E. Bet v 1 from birch pollen is a lipocalin-like protein acting as allergen only when devoid of iron by promoting Th2 lymphocytes. J. Biol. Chem. 2014;289:17416–17421. doi: 10.1074/jbc.M114.567875. PubMed DOI PMC

Roth-Walter F., Schmutz R., Mothes-Luksch N., Lemell P., Zieglmayer P., Zieglmayer R., Jensen-Jarolim E. Clinical efficacy of sublingual immunotherapy is associated with restoration of steady-state serum lipocalin 2 after SLIT: A pilot study. World Allergy Organ. J. 2018;11:21. doi: 10.1186/s40413-018-0201-8. PubMed DOI PMC

Stetefeld J., McKenna S.A., Patel T.R. Dynamic light scattering: A practical guide and applications in biomedical sciences. Biophys. Rev. 2016;8:409–427. doi: 10.1007/s12551-016-0218-6. PubMed DOI PMC

Tenopoulou M., Kurz T., Doulias P.T., Galaris D., Brunk U.T. Does the calcein-AM method assay the total cellular ‘labile iron pool’ or only a fraction of it? Biochem. J. 2007;403:261–266. doi: 10.1042/BJ20061840. PubMed DOI PMC

Novotna A., Pavek P., Dvorak Z. Novel Stably Transfected Gene Reporter Human Hepatoma Cell Line for Assessment of Aryl Hydrocarbon Receptor Transcriptional Activity: Construction and Characterization. Environ. Sci. Technol. 2011;45:10133–10139. doi: 10.1021/es2029334. PubMed DOI

Elkhatib N., Porshneva K., Montagnac G. Migration cues interpretation by clathrin-coated structures. Curr. Opin. Cell Biol. 2021;72:100–105. doi: 10.1016/j.ceb.2021.07.005. PubMed DOI

Wardlaw A.J., Rick E.M., Pur Ozyigit L., Scadding A., Gaillard E.A., Pashley C.H. New Perspectives in the Diagnosis and Management of Allergic Fungal Airway Disease. J. Asthma Allergy. 2021;14:557–573. doi: 10.2147/JAA.S251709. PubMed DOI PMC

Diaz P.I., Hong B.-Y., Dupuy A.K., Strausbaugh L.D. Mining the oral mycobiome: Methods, components, and meaning. Virulence. 2017;8:313–323. doi: 10.1080/21505594.2016.1252015. PubMed DOI PMC

Santus W., Devlin J.R., Behnsen J. Crossing Kingdoms: How the Mycobiota and Fungal-Bacterial Interactions Impact Host Health and Disease. Infect. Immun. 2021;89:1110–1128. doi: 10.1128/IAI.00648-20. PubMed DOI PMC

Gallelli B., Viviani M., Nebuloni M., Marzano A.V., Pozzi C., Messa P., Fogazzi G.B. Skin infection due to Alternaria species in kidney allograft recipients: Report of a new case and review of the literature. J. Nephrol. 2006;19:668–672. PubMed

Vermeire S.E., de Jonge H., Lagrou K., Kuypers D.R. Cutaneous phaeohyphomycosis in renal allograft recipients: Report of 2 cases and review of the literature. Diagn. Microbiol. Infect. Dis. 2010;68:177–180. doi: 10.1016/j.diagmicrobio.2010.06.002. PubMed DOI

Chen Z., Jiang Y., Wang D., Zheng M., Liu X., Yuan C. Enhancement in serum (1-3)-beta-D-glucan level by cutaneous alternariosis: A case report and literature review. Microb. Pathog. 2021;150:104703. doi: 10.1016/j.micpath.2020.104703. PubMed DOI

Gabriel M.F., Postigo I., Tomaz C.T., Martinez J. Alternaria alternata allergens: Markers of exposure, phylogeny and risk of fungi-induced respiratory allergy. Environ. Int. 2016;89–90:71–80. doi: 10.1016/j.envint.2016.01.003. PubMed DOI

Vailes L.D., Perzanowski M.S., Wheatley L.M., Platts-Mills T.A., Chapman M.D. IgE and IgG antibody responses to recombinant Alt a 1 as a marker of sensitization to Alternaria in asthma and atopic dermatitis. Clin. Exp. Allergy. 2001;31:1891–1895. doi: 10.1046/j.1365-2222.2001.00745.x. PubMed DOI

Lopes L., Borges-Costa J., Soares-Almeida L., Filipe P., Neves F., Santana A., Guerra J., Kutzner H. Cutaneous Alternariosis Caused by Alternaria infectoria: Three Cases in Kidney Transplant Patients. Healthcare. 2013;1:100–106. doi: 10.3390/healthcare1010100. PubMed DOI PMC

Kejik Z., Kaplanek R., Masarik M., Babula P., Matkowski A., Filipensky P., Vesela K., Gburek J., Sykora D., Martasek P., et al. Iron Complexes of Flavonoids-Antioxidant Capacity and Beyond. Int. J. Mol. Sci. 2021;22:646. doi: 10.3390/ijms22020646. PubMed DOI PMC

Chobot V., Hadacek F. Iron and its complexation by phenolic cellular metabolites: From oxidative stress to chemical weapons. Plant Signal. Behav. 2010;5:4–8. doi: 10.4161/psb.5.1.10197. PubMed DOI PMC

von Loetzen C.S., Hoffmann T., Hartl M.J., Schweimer K., Schwab W., Rosch P., Hartl-Spiegelhauer O. Secret of the major birch pollen allergen Bet v 1: Identification of the physiological ligand. Biochem. J. 2014;457:379–390. doi: 10.1042/BJ20130413. PubMed DOI

Jacob T., von Loetzen C.S., Reuter A., Lacher U., Schiller D., Schobert R., Mahler V., Vieths S., Rosch P., Schweimer K., et al. Identification of a natural ligand of the hazel allergen Cor a 1. Sci. Rep. 2019;9:8714. doi: 10.1038/s41598-019-44999-2. PubMed DOI PMC

Casanal A., Zander U., Dupeux F., Valpuesta V., Marquez J.A. Purification, crystallization and preliminary X-ray analysis of the strawberry allergens Fra a 1E and Fra a 3 in the presence of catechin. Acta Crystallogr. Sect. F Struct. Biol. Cryst. Commun. 2013;69:510–514. doi: 10.1107/S1744309113006945. PubMed DOI PMC

Vesic J., Stambolic I., Apostolovic D., Milcic M., Stanic-Vucinic D., Cirkovic Velickovic T. Complexes of green tea polyphenol, epigalocatechin-3-gallate, and 2S albumins of peanut. Food Chem. 2015;185:309–317. doi: 10.1016/j.foodchem.2015.04.001. PubMed DOI

Structure of PR 10 Allergen Ara h 8.01 with Quercetin. RCSB PDB. 2022. [(accessed on 19 January 2022)]. Available online: https://www.rcsb.org/structure/6B1D.

Hurlburt B.K., Offermann L.R., McBride J.K., Majorek K.A., Maleki S.J., Chruszcz M. Structure and function of the peanut panallergen Ara h 8. J. Biol. Chem. 2013;288:36890–36901. doi: 10.1074/jbc.M113.517797. PubMed DOI PMC

van Boxtel E.L., van den Broek L.A., Koppelman S.J., Vincken J.P., Gruppen H. Peanut allergen Ara h 1 interacts with proanthocyanidins into higher molecular weight complexes. J. Agric. Food Chem. 2007;55:8772–8778. doi: 10.1021/jf071585k. PubMed DOI

Ohradanova-Repic A., Prazenicova R., Gebetsberger L., Moskalets T., Skrabana R., Cehlar O., Tajti G., Stockinger H., Leksa V. Time to Kill and Time to Heal: The Multifaceted Role of Lactoferrin and Lactoferricin in Host Defense. Pharmaceutics. 2023;15:1056. doi: 10.3390/pharmaceutics15041056. PubMed DOI PMC

Li G., Wei Y., Ma L., Mao Y., Xun R., Deng Y. A novel highly sensitive soy aptasensor for antigen beta-conglycinin determination. Anal. Methods. 2021;13:3059–3067. doi: 10.1039/D1AY00701G. PubMed DOI

Chinnappan R., Rahamn A.A., AlZabn R., Kamath S., Lopata A.L., Abu-Salah K.M., Zourob M. Aptameric biosensor for the sensitive detection of major shrimp allergen, tropomyosin. Food Chem. 2020;314:126133. doi: 10.1016/j.foodchem.2019.126133. PubMed DOI

Tipu H.N., Ahmed D., Gardezi S.A.H. In silico identification of epitopes from house cat and dog proteins as peptide immunotherapy candidates based on human leukocyte antigen binding affinity. Iran J. Vet. Res. 2017;18:56–59. PubMed PMC

Wang X., Qiao O., Han L., Li N., Gong Y. A Novel Rabbit Anti-Myoglobin Monoclonal Antibody’s Potential Application in Rhabdomyolysis Associated Acute Kidney Injury. Int. J. Mol. Sci. 2023;24:7822. doi: 10.3390/ijms24097822. PubMed DOI PMC

Tang H., Fayomi A.P., Bai S., Gupta N., Cascio S., Yang D., Buckanovich R.J. Generation and characterization of humanized affinity-matured EGFL6 antibodies for ovarian cancer therapy. Gynecol. Oncol. 2023;171:49–58. doi: 10.1016/j.ygyno.2023.02.004. PubMed DOI PMC

Joyce A., Shea C., You Z., Gorovits B., Lepsy C. Determination of Anti-drug Antibody Affinity in Clinical Study Samples Provides a Tool for Evaluation of Immune Response Maturation. AAPS J. 2022;24:114. doi: 10.1208/s12248-022-00759-1. PubMed DOI PMC

Eberhardt J., Santos-Martins D., Tillack A.F., Forli S. AutoDock Vina 1.2.0: New Docking Methods, Expanded Force Field, and Python Bindings. J. Chem. Inf. Model. 2021;61:3891–3898. doi: 10.1021/acs.jcim.1c00203. PubMed DOI PMC

Trott O., Olson A.J. AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J. Comput. Chem. 2010;31:455–461. doi: 10.1002/jcc.21334. PubMed DOI PMC

Shahinozzaman M., Ishii T., Ahmed S., Halim M.A., Tawata S. A computational approach to explore and identify potential herbal inhibitors for the p21-activated kinase 1 (PAK1) J. Biomol. Struct. Dyn. 2020;38:3514–3526. doi: 10.1080/07391102.2019.1659855. PubMed DOI

Wang W., Liu R., Zhu Y., Wang L., Tang Y., Dou B., Tian S., Wang F. YuNu-Jian attenuates diabetes-induced cardiomyopathy: Integrating network pharmacology and experimental validation. Front. Endocrinol. 2023;14:1195149. doi: 10.3389/fendo.2023.1195149. PubMed DOI PMC

Kizhakedathil M.P.J., Madasu P.K., Chandran T., Vijaykumar S.D. In-silico structural studies on anti-inflammatory activity of phytocompounds from the genus Andrographis. J. Biomol. Struct. Dyn. 2023:1–13. doi: 10.1080/07391102.2023.2234486. PubMed DOI

Almansour N.M., Allemailem K.S., Abd El Aty A.A., Ismail E.I.F., Ibrahim M.A.A. In Silico Mining of Natural Products Atlas (NPAtlas) Database for Identifying Effective Bcl-2 Inhibitors: Molecular Docking, Molecular Dynamics, and Pharmacokinetics Characteristics. Molecules. 2023;28:783. doi: 10.3390/molecules28020783. PubMed DOI PMC

Xia J., Hu H., Xue W., Wang X.S., Wu S. The discovery of novel HDAC3 inhibitors via virtual screening and in vitro bioassay. J. Enzyme Inhib. Med. Chem. 2018;33:525–535. doi: 10.1080/14756366.2018.1437156. PubMed DOI PMC

Zeng L., Shin W.H., Zhu X., Park S.H., Park C., Tao W.A., Kihara D. Discovery of Nicotinamide Adenine Dinucleotide Binding Proteins in the Escherichia coli Proteome Using a Combined Energetic- and Structural-Bioinformatics-Based Approach. J. Proteome Res. 2017;16:470–480. doi: 10.1021/acs.jproteome.6b00624. PubMed DOI PMC

Tanchuk V.Y., Tanin V.O., Vovk A.I., Poda G. A New, Improved Hybrid Scoring Function for Molecular Docking and Scoring Based on AutoDock and AutoDock Vina. Chem. Biol. Drug Des. 2016;87:618–625. doi: 10.1111/cbdd.12697. PubMed DOI

Zhang Y., Gao Y., Liang Y., Dong Y., Yang X., Qiu D. Verticillium dahliae PevD1, an Alt a 1-like protein, targets cotton PR5-like protein and promotes fungal infection. J. Exp. Bot. 2019;70:613–626. doi: 10.1093/jxb/ery351. PubMed DOI PMC

Saenz-de-Santamaria M., Guisantes J.A., Martinez J. Enzymatic activities of Alternaria alternata allergenic extracts and its major allergen (Alt a 1) Mycoses. 2006;49:288–292. doi: 10.1111/j.1439-0507.2006.01238.x. PubMed DOI

Garrido-Arandia M., Tome-Amat J., Pazos-Castro D., Esteban V., Escribese M.M., Hernandez-Ramirez G., Yuste-Montalvo A., Barber D., Pacios L.F., Diaz-Perales A. Interaction of Alt a 1 with SLC22A17 in the airway mucosa. Allergy. 2019;74:2167–2180. doi: 10.1111/all.13877. PubMed DOI

Hayes T., Rumore A., Howard B., He X., Luo M., Wuenschmann S., Chapman M., Kale S., Li L., Kita H., et al. Innate Immunity Induced by the Major Allergen Alt a 1 From the Fungus Alternaria Is Dependent Upon Toll-like Receptors 2/4 in Human Lung Epithelial Cells. Front. Immunol. 2018;9:1507. doi: 10.3389/fimmu.2018.01507. PubMed DOI PMC

Lin L., Dai Y., Xia Y. An overview of aryl hydrocarbon receptor ligands in the Last two decades (2002–2022): A medicinal chemistry perspective. Eur. J. Med. Chem. 2022;244:114845. doi: 10.1016/j.ejmech.2022.114845. PubMed DOI

Koch S., Stroisch T.J., Vorac J., Herrmann N., Leib N., Schnautz S., Kirins H., Forster I., Weighardt H., Bieber T. AhR mediates an anti-inflammatory feedback mechanism in human Langerhans cells involving FcepsilonRI and IDO. Allergy. 2017;72:1686–1693. doi: 10.1111/all.13170. PubMed DOI

Aguilera-Montilla N., Chamorro S., Nieto C., Sanchez-Cabo F., Dopazo A., Fernandez-Salguero P.M., Rodriguez-Fernandez J.L., Pello O.M., Andres V., Cuenda A., et al. Aryl hydrocarbon receptor contributes to the MEK/ERK-dependent maintenance of the immature state of human dendritic cells. Blood. 2013;121:e108–e117. doi: 10.1182/blood-2012-07-445106. PubMed DOI

Maazi H., Shirinbak S., Bloksma N., Nawijn M.C., van Oosterhout A.J. Iron administration reduces airway hyperreactivity and eosinophilia in a mouse model of allergic asthma. Clin. Exp. Immunol. 2011;166:80–86. doi: 10.1111/j.1365-2249.2011.04448.x. PubMed DOI PMC

Leung S., Holbrook A., King B., Lu H.T., Evans V., Miyamoto N., Mallari C., Harvey S., Davey D., Ho E., et al. Differential inhibition of inducible T cell cytokine secretion by potent iron chelators. J. Biomol. Screen. 2005;10:157–167. doi: 10.1177/1087057104272295. PubMed DOI

Jason J., Archibald L.K., Nwanyanwu O.C., Bell M., Jensen R.J., Gunter E., Buchanan I., Larned J., Kazembe P.N., Dobbie H., et al. The effects of iron deficiency on lymphocyte cytokine production and activation: Preservation of hepatic iron but not at all cost. Clin. Exp. Immunol. 2001;126:466–473. doi: 10.1046/j.1365-2249.2001.01707.x. PubMed DOI PMC

Thorson J.A., Smith K.M., Gomez F., Naumann P.W., Kemp J.D. Role of iron in T cell activation: TH1 clones differ from TH2 clones in their sensitivity to inhibition of DNA synthesis caused by IgG Mabs against the transferrin receptor and the iron chelator deferoxamine. Cell Immunol. 1991;134:126–137. doi: 10.1016/0008-8749(91)90336-A. PubMed DOI

Drury K.E., Schaeffer M., Silverberg J.I. Association between Atopic Disease and Anemia in US Children. JAMA Pediatr. 2016;170:29–34. doi: 10.1001/jamapediatrics.2015.3065. PubMed DOI

Rhew K., Brown J.D., Oh J.M. Atopic Disease and Anemia in Korean Patients: Cross-Sectional Study with Propensity Score Analysis. Int. J. Environ. Res. Public Health. 2020;17:1978. doi: 10.3390/ijerph17061978. PubMed DOI PMC

Rhew K., Oh J.M. Association between atopic disease and anemia in pediatrics: A cross-sectional study. BMC Pediatr. 2019;19:455. doi: 10.1186/s12887-019-1836-5. PubMed DOI PMC

Shaheen S.O., Gissler M., Devereux G., Erkkola M., Kinnunen T.I., McArdle H., Sheikh A., Hemminki E., Nwaru B.I. Maternal iron supplementation in pregnancy and asthma in the offspring: Follow-up of a randomised trial in Finland. Eur. Respir. J. 2020;55:1902335. doi: 10.1183/13993003.02335-2019. PubMed DOI

Shaheen S.O., Macdonald-Wallis C., Lawlor D.A., Henderson A.J. Haemoglobin concentrations in pregnancy and respiratory and allergic outcomes in childhood: Birth cohort study. Clin. Exp. Allergy. 2017;47:1615–1624. doi: 10.1111/cea.13034. PubMed DOI PMC

Shaheen S.O., Newson R.B., Henderson A.J., Emmett P.M., Sherriff A., Cooke M., Team A.S. Umbilical cord trace elements and minerals and risk of early childhood wheezing and eczema. Eur. Respir. J. 2004;24:292–297. doi: 10.1183/09031936.04.00117803. PubMed DOI

Nyakeriga A.M., Williams T.N., Marsh K., Wambua S., Perlmann H., Perlmann P., Grandien A., Troye-Blomberg M. Cytokine mRNA expression and iron status in children living in a malaria endemic area. Scand J. Immunol. 2005;61:370–375. doi: 10.1111/j.1365-3083.2005.01573.x. PubMed DOI

Ghatak S.K., Majumdar D., Singha A., Sen S., Das D., Chakrabarti A., Mukhopadhyay C., Sen K. Peanut protein sensitivity towards trace iron: A novel mode to ebb allergic response. Food Chem. 2015;176:308–313. doi: 10.1016/j.foodchem.2014.12.081. PubMed DOI

Tong P., Gao L., Gao J., Li X., Wu Z., Yang A., Chen H. Iron-induced chelation alleviates the potential allergenicity of ovotransferrin in a BALB/c mouse model. Nutr. Res. 2017;47:81–89. doi: 10.1016/j.nutres.2017.09.009. PubMed DOI

Word J.M., Lovell S.C., Richardson J.S., Richardson D.C. Asparagine and glutamine: Using hydrogen atom contacts in the choice of side-chain amide orientation. J. Mol. Biol. 1999;285:1735–1747. doi: 10.1006/jmbi.1998.2401. PubMed DOI

Morris G.M., Huey R., Lindstrom W., Sanner M.F., Belew R.K., Goodsell D.S., Olson A.J. AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility. J. Comput. Chem. 2009;30:2785–2791. doi: 10.1002/jcc.21256. PubMed DOI PMC

Pettersen E.F., Goddard T.D., Huang C.C., Couch G.S., Greenblatt D.M., Meng E.C., Ferrin T.E. UCSF Chimera--a visualization system for exploratory research and analysis. J. Comput. Chem. 2004;25:1605–1612. doi: 10.1002/jcc.20084. PubMed DOI

Borrego J., Naseem M.U., Sehgal A.N.A., Panda L.R., Shakeel K., Gaspar A., Nagy C., Varga Z., Panyi G. Recombinant Expression in Pichia pastoris System of Three Potent Kv1.3 Channel Blockers: Vm24, Anuroctoxin, and Ts6. J. Fungi. 2022;8:1215. doi: 10.3390/jof8111215. PubMed DOI PMC

Hufnagl K., Afify S.M., Braun N., Wagner S., Wallner M., Hauser M., Wiederstein M., Gadermaier G., Wildner S., Redegeld F.A., et al. Retinoic acid-loading of the major birch pollen allergen Bet v 1 may improve specific allergen immunotherapy: In silico, in vitro and in vivo data in BALB/c mice. Allergy. 2020;75:2073–2077. doi: 10.1111/all.14259. PubMed DOI PMC

Roth-Walter F., Starkl P., Zuberbier T., Hummel K., Nobauer K., Razzazi-Fazeli E., Brunner R., Pali-Scholl I., Kinkel J., Felix F., et al. Glutathione exposes sequential IgE-epitopes in ovomucoid relevant in persistent egg allergy. Mol. Nutr. Food Res. 2013;57:536–544. doi: 10.1002/mnfr.201200612. PubMed DOI PMC

Oida K., Einhorn L., Herrmann I., Panakova L., Resch Y., Vrtala S., Hofstetter G., Tanaka A., Matsuda H., Jensen-Jarolim E. Innate function of house dust mite allergens: Robust enzymatic degradation of extracellular matrix at elevated pH. World Allergy Organ. J. 2017;10:23. doi: 10.1186/s40413-017-0154-3. PubMed DOI PMC