Cloning, Isolation, and IgE-Binding Properties of &lt;i&gt;Helix aspersa&lt;/i&gt; (Brown Garden Snail) Tropomyosin

Asturias, Juan A.; Eraso, Elena; Arilla, Maite; Gómez-Bayón, N; Inácio, Filipe; Martı́nez, Alberto

doi:10.1159/000059398

Cited by 44 publications

(33 citation statements)

References 12 publications

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“…Also, with invertebrates such as crustaceans (shrimps, crabs, lobsters and crawfish), insects (cockroaches) and mollusks (squid, mussel, clam and snail), adverse food reactions have been found in allergic individuals due to the muscle protein tropomyosin. Sequence identities and similarities between H. aspersa and D. pteronyssinus tropomyosin are 65 and 79, respectively [40], but a 36-kDa protein from the H. aspersa extract, and the natural and recombinant purified tropomyosins were recognized by IgE antibodies in only 4 of 22 sera from patients reporting snail reactivity and showing specific IgE to H. aspersa above class 2 [41]. In our study, a 37-kDa protein from H. aspersa extract was also recognized by the IgE in only 1 out of 21 patients, and the two major allergens found are probably the heavy chains of myosin (∼225 kDa), which were found in 13 and 18 of 21 patients, respectively.…”

Section: Discussionmentioning

confidence: 99%

The Helix aspersa (Brown Garden Snail) Allergen Repertoire

Martins¹,

Peltre²,

Faro³

et al. 2005

Int Arch Allergy Immunol

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Background: Ingestion of snails can induce strong asthmatic or anaphylactic responses, mainly in house-dust-mite-sensitized patients. The aim of this study was to identify the Helix aspersa (Hel a), Theba pisana (The p) and Otala lactea (Ota l) allergens and the extent of their cross-reactivity with the Dermatophagoides pteronyssinus (Der p) mite. Patients and Methods: In 60 atopicpatients, skin prick tests (SPT) to snail and D. pteronyssinus, total and specific IgE, specific IgE immunoblots, RAST and immunoblot inhibition assays were performed. Results: Mean total IgE was >1,000 kU/l. Mean specific IgE (class 6 for Der p and class 2 for Hel a) SPT were positive in 44 patients for snail and in 56 for mite. Isoelectric focusing (IEF) and SDS-PAGE followed by immunoblotting of H. aspersa extract enabled the identification of 27 and 20 allergens, respectively. Myosin heavy chains from snails (molecular weight >208 kDa) disclosed two major allergens. Hel a and Der p RAST were strongly inhibited by their homologous extracts, with Hel a RAST being inhibited by the Der p extract to a much greater extent (72.6%) than the inverse (5.6%). A complete inhibition of the immunoblots by their homologous extract was obtained. However, Hel a extract did not inhibit Der p IEF separated recognition. On the other hand, mite extract extensively inhibited snail immunoblots from both IEF and SDS-PAGE separations. Immune detection on chicken, pig, rabbit, cow and horse myosins did not reveal any IgE cross recognition with snail. Conclusions: In most cases of snail allergy, mite appeared to be the sensitizing agent. Nevertheless, snails may also be able to induce sensitization by themselves. This hypothesis is supported by the finding of specific IgE to Hel a in 2 patients who did not show specific IgE to Der p, and one of them was suffering from asthma after snail ingestion.

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Section: Discussionmentioning

confidence: 99%

The Helix aspersa (Brown Garden Snail) Allergen Repertoire

Martins¹,

Peltre²,

Faro³

et al. 2005

Int Arch Allergy Immunol

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“…Chemiluminescence was detected using GeneGnome apparatus (Syngene, Cambridge, UK). Otherwise, for the detection of tropomyosin, the electrotransferred and blocked membranes were incubated with 1:5,000 diluted polyclonal antibodies obtained from rabbits immunized with shrimp tropomyosin (Pen m 1; Bial Aristegui, Lisbon, Portugal), that can also bind to tropomyosins from other sources [12,25,26], and 1:2,000 diluted HRP-conjugated anti-rabbit IgG (Sigma-Aldrich, St. Louis, Mo., USA). Detection of bands was performed as above.…”

Section: Methodsmentioning

confidence: 99%

Identification and Characterization of IgE-Binding Tropomyosins in Aedes aegypti

Cantillo

Puerta

Lafosse-Marin³

et al. 2016

Int Arch Allergy Immunol

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Background: The mosquito Aedes aegypti is a potential source of important clinically relevant allergens. However, the allergenicity and cross-reactivity of most of these has not been fully described. Methods: Natural wild-type mosquito tropomyosin was purified by size exclusion and anionic-exchange chromatography from an A. aegypti extract. Further characterization was accomplished by MALDI-TOF/TOF. Two recombinant variants of tropomyosin were obtained by expression in Escherichia coli. Specific IgE measurement by ELISA and skin tests for mosquito extract were performed in 12 patients with asthma or allergy rhinitis residing on the Caribbean island of Martinique. Cross-reactivity between natural A. aegypti tropomyosin and recombinant tropomyosins from A. aegypti, house dust mite, shrimp and Ascaris lumbricoides was analyzed by ELISA competition. Results: Four variants of natural tropomyosin were purified. A band of 32 kDa in SDS-PAGE representing 2 tropomyosin variants (Aed a 10.0101 and Aed a 10.0201) reacted with specific IgE of 4 of the 12 (33%) allergic patients and with rabbit polyclonal anti-shrimp tropomyosin. A high degree of cross-reactivity (60-70%) was detected between natural mosquito tropomyosin and Blo t 10, Der p 10 and Lit v 1, and a lower degree with Asc l 3 from A. lumbricoides (<30%). rAed a 10.0101 inhibited IgE binding to natural A. aegypti tropomyosin; however, rAed a 10.0201 showed a low inhibitory capacity. Conclusion: Tropomyosin is a new IgE-binding protein from A. aegypti. Two of the 4 variants identified showed different degree of cross-reactivity with tropomyosins from other arthropods. The potential allergenic role of each variant should be further investigated.

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“…Tropomyosin is the most widely reported allergen in mollusks [4] and has been identified in cephalopods [25,26], bivalves [27,28], and gastropods [29]. It should be noted that in some of these studies, the sera used came from patients who were allergic to crustaceans.…”

Section: Discussionmentioning

confidence: 99%

Heterogeneity in Allergy to Mollusks: A Clinical-Immunological Study in a Population From the North of Spain

Azofra

Echechipía

Irazábal

et al. 2017

J Investig Allergol Clin Immunol

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Background: Allergy to mollusks has been the focus of fewer studies than allergy to crustaceans. Furthermore, allergy to mollusks is less well characterized. Objectives: To describe the clinical characteristics of mollusk-allergic patients, to identify the responsible allergens, and to assess crossreactivity. Methods: We performed a prospective multicenter study including 45 patients with mollusk allergy, which was diagnosed based on a suggestive clinical history and a positive skin test result with the agent involved. Fractions were identified using SDS-PAGE and immunoblotting. The proteins responsible were subsequently identified using mass spectrometry. ELISA inhibition studies were performed with mollusks, dust mites, and crustaceans. Results: We found that 25 patients (55%) were allergic to cephalopods, 14 (31%) to bivalves, and 11 (24%) to gastropods. Limpet was the third most frequent cause of allergy (15% of cases). In 31 patients (69%), the manifestation was systemic; 10 (22%) exhibited oral allergy syndrome, and 7 (15%) experienced contact urticaria. Most major allergens were found between 27 kDa and 47 kDa. ELISA inhibition assays revealed a high degree of inhibition of cephalopods and bivalves from all the groups of mollusks, mites, and crustaceans. Mass spectrometry identified tropomyosin, actin, and myosin as the major allergens. Conclusions: Cephalopods, especially squid, are the mollusks that most frequently trigger allergic symptoms. The very frequent occurrence of allergy to limpets is striking, given their low consumption in our area. It is worth highlighting the heterogeneity observed, exemplified by the gastropods. Tropomyosin appears to be responsible for the high cross-reactivity found between mollusks, mites, and crustaceans. Three new mollusk allergens were also identified, namely, actin, enolase, and a putative C1q domain-containing protein.

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Cloning, Isolation, and IgE-Binding Properties of <i>Helix aspersa</i> (Brown Garden Snail) Tropomyosin

Cited by 44 publications

References 12 publications

The <i>Helix aspersa</i> (Brown Garden Snail) Allergen Repertoire

The <i>Helix aspersa</i> (Brown Garden Snail) Allergen Repertoire

Identification and Characterization of IgE-Binding Tropomyosins in <b><i>Aedes aegypti</i></b>

Heterogeneity in Allergy to Mollusks: A Clinical-Immunological Study in a Population From the North of Spain

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