Person: Cuéllar Del Hoyo, María Del Carmen
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First Name
María Del Carmen
Last Name
Cuéllar Del Hoyo
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Farmacia
Department
Microbiología y Parasitología
Area
Parasitología
Identifiers
7 results
Search Results
Now showing 1 - 7 of 7
Item Are fish tropomyosins allergens?(Annals of Allergy, Asthma & Immunology, 2016) Veleiro, Beatriz; Daschner, Alvaro; González Fernández, Juan; Cuéllar Del Hoyo, María Del CarmenItem Possible Allergenic Role of Tropomyosin in Patients with Adverse Reactions after Fish Intake(Immunological Investigations, 2018) Alguacil-Guillén, Marina; Daschner, Alvaro; González Fernández, Juan; Cuéllar Del Hoyo, María Del CarmenIn a recent case report, patient's anti-fish tropomyosin IgE was associated with gastrointestinal symptoms. We aimed to demonstrate on a wider scale that the panallergen tropomyosin should not be limited to invertebrate species and that clinically relevant reactions could be elicited by vertebrate tropomyosin. On the whole, 19 patients with adverse reactions after fish intake and showing negative skin tests with commercial fish extracts were included. Fish tropomyosin was recognized by 10/19 patients' IgE by immunoblotting. All patients with gastrointestinal complaints after fish intake (6/6) showed an IgE band matching with tropomyosin. Cod, albacore, and swordfish tropomyosins were recognized by most patients although 3/10 patients did not claim adverse reactions to these fish species. Immunoblotting with a battery of antigens from different fish species have a high yield of positivity at a band matching with tropomyosin molecular weight, even if they have not been claimed to be causative agents of symptoms. Tropomyosin is therefore a good candidate to be investigated as a clinically relevant fish allergen in patients who report adverse reactions after fish intake.Item New insights into the allergenicity of tropomyosin: a bioinformatics approach(Molecular Biology Reports, 2014) González Fernández, Juan; Rodero Martínez, Marta; Daschner, Álvaro; Cuéllar Del Hoyo, María Del CarmenThe invertebrate panallergen tropomyosin is a protein with an extremely simple folding. This makes it a perfect target for investigating structural differences between invertebrate and vertebrate tropomyosins, which are not considered allergenic. Phylogenetic and sequence analyses were conducted in order to explore the differences in primary structure between several tropomyosins and to promote an experimental development in the field of food allergy, based on the study of tropomyosin. The phylogenetic analyses showed that tropomyosin is a useful evolutionary marker. The phylogenetic trees obtained with tropomyosin were not always phylogenetically correct, but they might be useful for allergen avoidance by tropomyosin allergic individuals. Sequence analyses revealed that the probability of alpha helix folding in invertebrate tropomyosins was lower than in all the studied vertebrate ones, except for the Atlantic bluefin tuna Thunnus thynnus tropomyosin. This suggested that the lack of alpha helix folding may be involved in the immunogenicity of tropomyosins. More specifically, the regions adjacent to the positions 133-135 and 201 of the invertebrate tropomyosins, presented lower probability of alpha helix folding than those of vertebrates and are candidates to be responsible for their allergenicity.Item Diamine oxidase levels in different chronic urticaria phenotypes(Allergologia et Immunopathologia, 2015) Alvaro Daschner; Juan González-Fernández; Ana Valls; Consolación de Frutos; Marta Rodero; González Fernández, Juan; Cuéllar Del Hoyo, María Del Carmen; Rodero Martínez, MartaBackground: Diamine oxidase (DAO) is a polyamine-degrading enzyme also implicated in histamine metabolism. Chronic urticaria (CU) has a wide spectrum of clinical presentations and causes. Anisakis sensitisation associated chronic urticaria (CU+) has been characterised as a phenotype with different clinical and immunological characteristics and possibly associated with previous acute parasitism. We aimed to analyse serum DAO levels in different CU phenotypes. We further analysed the possible association of DAO with fish eating habits. Methods: We studied 35 CU+ patients and 39 non-sensitised CU patients (CU-) as well as 19 controls. We analysed fish-eating frequency as well as fish intake associated exacerbation of CU (FIAE) or gastro-intestinal complaints (GI). DAO levels were further analysed with respect to lymphoproliferative responses, cytokine and specific IgE production. Results: DAO levels were not different between CU and controls, but were significantly higher in CU+ than in CU-. CU+ patients with FIAE had lower DAO levels, but no differences were detected in patients with GI. DAO levels correlated positively with oily and canned fish consumption in CU-. In CU+, DAO levels correlated positively with specific Anisakis IgE, percentages of proliferation in Anisakis stimulated peripheral blood lymphocytes, serum IL-2 and IL-6, but correlated negatively with mitogen stimulated TGF-β in supernatants. Conclusions: DAO levels in CU depend on fish-eating habits and in CU+ on the amount of specific IgE production. In the CU+ phenotype, lower levels of DAO predispose to urticaria exacerbation after fish intake, probably due to a relative insufficient enteric availability of this enzyme.Item Allergenicity of vertebrate tropomyosins: Challenging an immunological dogma(Allergologia et Immunopathologia, 2017) Daschner, A.; González Fernández, Juan; Cuéllar Del Hoyo, María Del CarmenWith the exception of tilapia tropomyosin, other anecdotic reports of tropomyosin recognition of vertebrate origin are generally not accompanied by clinical significance and a dogmatic idea is generally accepted about the inexistence of allergenicity of vertebrate tropomyosins, based mainly on sequence similarity evaluations with human tropomyosins. Recently, a specific work-up of a tropomyosin sensitised patient with seafood allergy, demonstrated that the IgE-recognition of tropomyosin from different fish species can be clinically relevant. We hypothesise that some vertebrate tropomyosins could be relevant allergens. The hypothesis is based on the molecular evolution of the proteins and it was tested by in silico methods. Fish, which are primitive vertebrates, could have tropomyosins similar to those of invertebrates. If the hypothesis is confirmed, tropomyosin should be included in different allergy diagnosis tools to improve the medical protocols and management of patients with digestive or cutaneous symptoms after fish intake.Item Haemoglobin, a new major allergen of Anisakis simplex(International Journal for Parasitology, 2015) Daschner, Alvaro; Nieuwenhuizen, Natalie E.; Lopata, Andreas L.; Frutos, Consolación De; Valls, Ana; González Fernández, Juan; Cuéllar Del Hoyo, María Del CarmenGastro-allergic anisakiasis and Anisakis sensitisation associated chronic urticaria are diseases which differ in their IgE and IgG4 responses against both crude extract and specific allergens. Anisakis and Ascaris are closely related nematodes that usually cause problems with specificity in immunodiagnostics. In this study we measured IgE and IgG4 antibodies against Anisakis simplex sensu lato (s. l.) and Ascaris suum haemoglobins in sera of 21 gastro-allergic anisakiasis and 23 chronic urticaria patients. We used a capture ELISA with the anti-Anisakis haemoglobin monoclonal antibody 4E8g, which also recognises Ascaris haemoglobin. In addition, we determined specific IgE and IgG4 to both nematodes by indirect ELISA and immunoblotting. Anti-A. simplex s. l. haemoglobin IgE and IgG4 levels were higher in gastro-allergic anisakiasis than in chronic urticaria patients (P=0.002 and 0.026, respectively). Surprisingly, no patient had detectable IgE levels against A. suum haemoglobin. Finally, we carried out an in silico study of the B-cell epitopes of both haemoglobin molecules. Five epitopes were predicted in Anisakis pegreffii and four in A. suum haemoglobin. The epitope propensity values of Anisakis haemoglobin in the equivalent IgE binding region of the allergenic haemoglobin Chi t 1 from Chironomus thummi, were higher those of the Ascaris haemoglobin. In conclusion, we describe A. simplex haemoglobin as a new major allergen (Ani s 13), being recognised by a large number (64.3%) of sensitised patients and up to 80.9% in patients with gastro-allergic anisakiasis. The presence of a specific epitope and the different values of epitope propensity between Anisakis and Ascaris haemoglobin could explain the lack of cross-reactivity between the two molecules. The absence of IgE reactivity to Ascaris haemoglobin in Anisakis patients makes Anisakis haemoglobin (Ani s 13) a potential candidate for developing more specific diagnosis tools.Item Recombinant vs native Anisakis haemoglobin (Ani s 13): its appraisal as a new gold standard for the diagnosis of allergy(Experimental Parasitology, 2017) Rivas, Luis; Luque-Ortega, Juan Román; Núñez-Ramírez, Rafael; Campioli, Pamela; Gárate, Teresa; Perteguer, María J.; Daschner, Alvaro; González Fernández, Juan; Cuéllar Del Hoyo, María Del CarmenRecombinant allergens are currently the best option for serodiagnosis of human anisakiasis in terms of sensitivity and specificity. However, previous reports showed high rates of anisakiasis patients who were negative to Ani s 7 and especially to Ani s 1. Recently, Anisakis haemoglobin was described as a major allergen (Ani s 13). Although Ani s 13 belongs to a conserved protein family, it seems not to be a cross-reacting antigen because of the absence of IgE recognition against Ascaris haemoglobin in Anisakis patients. The aim of this study is to develop a more sensitive and specific diagnosis tool for Anisakis based on the recently discovered allergen Ani s 13. We obtained and purified recombinant Anisakis haemoglobin (rAni s 13) and the native form (nAni s 13). The recognition of both recombinant and native haemoglobins by anti-haemoglobin IgE from patients' sera was assessed by indirect ELISA and immunoblotting using 43 Anisakis sensitised patients and 44 non-Anisakis sensitised patients. Native Ani s 13 was also treated with periodate to study if oxidation of glycans destroys antibody binding. Furthermore, it was structurally characterised by negative staining electron microscopy and analytical ultracentrifugation. Recombinant Ani s 13 was only recognised by four patients with gastro-allergic anisakiasis (GAA) and immunoblotting analyses showed no bands. However, nAni s 13 was detected by 72.1% of Anisakis sensitised patients measured by indirect ELISA. Particularly, 18 (90%) out of 20 GAA patients were positive. Tetramers and octamers were the most abundant homomers of nAni s 13 but octamers had higher content of bound heme. None of the non-Anisakis sensitised patients were positive. Combined use of purified native form of Ani s 13 with current gold standards would improve the sensitivity and specificity for diagnosing anisakiasis.