Alternaria alternata is the most important allergenic fungus, with up to 20% of allergic patients affected. The sensitization profile of patients sensitized to A. alternata and how it changes when treated with immunotherapy is not known. Our objective is to determine the allergen recognition pattern of allergic patients to A. alternata and to study its association to the parameters studied in a clinical trial recently published. Sera of 64 patients from the clinical trial of immunotherapy with native major allergen Alt a 1 were analyzed by immunoblotting; 98. 4% of the patients recognized Alt a 1. The percentage of recognition for Alt a 3, Alt a 4, and/or Alt a 6, Alt a 7, Alt a 8, Alt a 10 and/or Alt a 15 was 1.6%, 21.9%, 12.5%, 12.5%, and 12.5% respectively. Of the 64 patients, 45 (70.3%) only recognized Alt a 1 among the allergens present in the A. alternata extract. Immunotherapy with Alt a 1 desensitizes treated patients, reducing their symptoms and medication consumption through the elimination of Alt a 1 sensitization, which is no longer present in the immunoblotting of some patients. There may be gender differences in the pattern of sensitization to A. alternata allergens, among others.

Orange allergy is estimated to account for up to 3–4% of food allergies. Major allergens identified in orange (Citrus sinensis) include Cit s 1 (germin-like protein) and Cit s 2 (profilin), while Cit s 3 (non-specific lipid transfer protein, nsLTP) and Cit s 7 (gibberellin-regulated protein) have also been described. The objective of this study was to investigate the presence and IgE-binding capacity of germin-like proteins in citrus fruits other than oranges. We describe five patients with immediate allergic reactions after orange ingestion. All patients underwent skin prick tests (SPT) to aeroallergens and common food allergens, prick-by-prick testing with orange, lemon, and mandarin (pulp, peel, seeds), total IgE, specific IgE (sIgE), anaphylaxis scoring (oFASS), and the Food Allergy Quality of Life Questionnaire (FAQLQ-AF). Protein extracts from peel and pulp of orange, lemon, and mandarin were analyzed by Bradford assay, SDS-PAGE, and IgE immunoblotting using patient sera. Selected bands were identified by peptide mass fingerprinting. A 23 kDa band was recognized by all five patients in orange (pulp and peel), lemon (peel), and mandarin (peel). This band was consistent with Cit s 1, a germin-like protein already annotated in the IUIS allergen database for orange but not for lemon or mandarin. Peptide fingerprinting confirmed the germin-like identity of the 23 kDa bands in all three citrus species. Germin-like proteins of approximately 23 kDa were identified as IgE-binding components in peel extracts of orange, lemon, and mandarin, and in orange pulp. These findings suggest a potential shared allergen across citrus species that may contribute to allergic reactions independent of LTP sensitization.

Background/Objectives: The aim of this study was to evaluate the tolerability and effectiveness of subcutaneous immunotherapy (SCIT) in allergic adults and children treated with a polymerized-glutaraldehyde undiluted mixture of house dust mites (HDMs) under routine clinical practice. Methods: This was an observational, ambispective, controlled, real-world, multicenter study including patients ≥ 5 years with allergic rhinitis (AR), due to Dermatophagoides sensitization. Patients who started AIT with a D. pteronyssinus/D. farinae extract and those who continued symptomatic treatment were included in the treatment (DP&DF) and untreated (UT) groups, respectively. We evaluated adverse reactions (ARs) and changes in effectiveness variables through changes in symptoms, disease control, medication use, and patient- and investigator-reported outcomes. Results: We included 130 patients in the DP&DF group, and 90 (69.2%) adults, 23 adolescents (17.7%), 17 (13.1%) children, and 94 patients in the UT group. Patients received treatment for a mean (SD) of 9.01 (3.1) months at the time of evaluation. Seven (5.4%) patients, all adults, reported eight ARs, five local and three systemic (mean rate of 0.62 ARs per 100 injections); all recovered, and epinephrine was not required. The proportion of patients reporting no rhinitis symptoms at follow-up significantly increased (+13.6%; p < 0.001). Rhinitis frequency, intensity, and control significantly improved overall and in specific age groups. Similarly, the proportion of patients reporting no asthma symptoms at follow-up significantly increased (+29.0%; p < 0.001). The use of all symptomatic medications significantly decreased, while the UT group showed no significant changes, except for worsened asthma classification and control in specific age groups. Both investigators and patients perceived a marked improvement in symptoms and medication use, with high satisfaction scores reported on the visual analogue scale. Conclusions: A subcutaneous allergen extract with a mixture of HDMs is safe and effective for allergic rhinitis and asthma in adults and children in the real-world setting.

Between 1% and 2% of the world’s population is sensitised to mites. Aetiological diagnosis is key to the management of allergic patients. However, methods based solely on morphological criteria are ambiguous in many cases. Polymerase chain reaction of ribosomal genes represents a valuable complementary approach. The 5 most representative species (Dermatophagoides pteronyssinus, Dermatophagoides farinae, Tyrophagus putrescentiae, Blomia tropicalis and Lepidoglyphus destructor) were selected as sources of allergens. They were first identified morphologically and the 18S rRNA gene sequences were obtained from the GenBank database. Alignment of the nucleotide sequences of the 18S rRNA ribosomal gene enabled the identification of the conserved and divergent regions in all of them. The alignment allowed the design of a pair of oligonucleotides in conserved regions of the gene, to amplify the sequence of interest in each of the species. We performed genomic DNA extraction, quantification and purity. PCR, using oligonucleotides designed to amplify the 18S sequence fragment of interest, showed the exact size for each species. Amplification, efficiency curves and melting points resulting from the amplification of the 18S amplicon of the five species were obtained. The oligonucleotides designed for real-time PCR studies, allow species identification by amplifying the specific fragment of each species using real-time PCR.

House dust mite allergens Der p 1, Der p 2, and Der p 23 are recognized as major clinically relevant allergens worldwide; however, it is difficult to obtain these proteins in purified form from a natural source, which limits their use in molecular targeted immunotherapy and in vivo diagnosis. In this study, we developed and validated robust methodologies for the large-scale purification and individual characterization of native nDer p 1, nDer p 2, and nDer p 23 allergens from the natural sensitization source, Dermatophagoides pteronyssinus. Each allergen was isolated through an independent downstream process based on successive chromatographic steps, achieving high purity and preserving the structural integrity. Molecular standardization was performed in vivo in 27 mite-allergic patients by skin prick testing (SPT), enabling the separate determination of histamine equivalent potency (HEP) values: 7.43 µg/mL for nDer p 1, 8.11 µg/mL for nDer p 2, and 1.55 µg/mL for nDer p 23. These data establish a direct relationship between the protein concentration and biological activity for each major allergen. In conclusion, the successful production and biological standardization of native nDer p 1, nDer p 2, and nDer p 23 proteins provide well-defined reagents for in vivo molecular diagnosis and enable more precise and reproducible standardization compared with complex allergen extracts.

Orange allergy is estimated to account for 3% of food allergies. Cit s 1 (germin-like protein) and Cit s 2 (profilin) have been identified as major allergens. Also, Cyt s 3 (lipid transfer protein) and Cyt s 7 (gibberellin-regulated protein) have been described as orange allergens. This study focuses on the unusual occurrence of orange allergy in five patients. Materials and methods: We report five cases of patients with immediate allergic manifestations after ingestion of orange. Skin prick-tests to common aeroallergens, and different foods. Prick by prick with orange, lemon pulp mandarin (peel, pulp and seeds). Anaphylaxis score; Total IgE, Serum-specific IgE and food allergy quality of life questionnaire. Allergenic extracts were prepared from orange (Citrus sinensis), lemon (Citrus lemon) and mandarin (Citrus reticulata) peel and pulp. Protein characterization was determined by Bradford and SDS-PAGE and sera by immunoblot. Selected proteins were identified by peptide fingerprinting. Results: A 23 kD band was recognized by all 5 patients in orange (pulp and peel), lemon (peel) and mandarin (pulp and peel), having already been annotated in IUIS in the case of orange as germin (Cit s 1). To send the 23 kDa bands for peptide fingerprinting, SDS-PAGE of the 3 fruits was performed and as a positive control we used the 23 kDa band from orange. Together, the 23 kDa bands from orange, lemon and mandarin peel were cut out and sent for peptide fingerprint analysis, yielding the germin like protein sequence in all cases. Conclusions: A band of 23 kDa (germin-like protein) appears common in orange (annotated in IUIS as germin), lemon and tange-rine peel. Not previously described in IUIS in lemon and mandarin and confirmed as germin. Germin was in the peel of the 3 fruits and in the orange pulp, but not in lemon or mandarin.