RESUMO
BACKGROUND: Alfalfa (Medicago sativa L) is one of the most planted crops worldwide primarily used to feed animals. The use of alfalfa in human diet as sprouts, infusions and nutritional supplements is rapidly gaining popularity. Despite this, allergenicity assessment of this novel plant food is largely lacking. RESULTS: Here, leaf protein extract of alfalfa was studied using a combined proteomics, Immunoglobulin E (IgE)-binding inhibition assay and in silico approach to find potential allergens. We have identified and annotated 129 proteins using in-gel digestion proteomics and Blast2Go suit. A search against COMPARE database, using the identified proteins as query sequences, revealed high similarity with several allergenic proteins. The Single Point Highest Inhibition Achievable assay (SPHIAa) performed on the multiplex FABER® allergy testing system confirmed the in silico results and showed some additional potential allergens. This approach allowed the detection of proteins in alfalfa leaves cross-reacting with plant allergens from three different allergen families such as lipid transfer, thaumatin-like and Bet v 1-like protein families. In addition, the absence of structural determinants cross-reacting with seed storage allergenic proteins and with animal allergens was recorded. CONCLUSION: This study reports for the first time potential allergenic proteins in alfalfa. The results suggest that this plant food can be safely introduced, as a protein-rich supplement, in the diet of patients allergic to animal food allergens. Allergic patients towards certain plant food allergens need to be careful about consuming alfalfa because they might have allergic symptoms. © 2020 Society of Chemical Industry.
Assuntos
Alérgenos/imunologia , Imunoglobulina E/imunologia , Medicago sativa/imunologia , Alérgenos/química , Alérgenos/genética , Sequência de Aminoácidos , Simulação por Computador , Reações Cruzadas , Medicago sativa/química , Medicago sativa/genética , Folhas de Planta/química , Folhas de Planta/genética , Folhas de Planta/imunologia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , ProteômicaRESUMO
BACKGROUND: Kiwifruit is an important cause of food allergy. A high amount of a protein with a molecular mass compatible with that of Bet v 1 was observed in the kiwifruit extract. OBJECTIVE: To identify and characterize kirola, the 17-kDa protein of green kiwifruit (Act d 11). METHODS: Act d 11 was purified from green kiwifruit. Its primary structure was obtained by direct protein sequencing. The IgE binding was investigated by skin testing, immunoblotting, inhibition tests, and detection by the ISAC microarray in an Italian cohort and in selected Bet v 1-sensitized Austrian patients. A clinical evaluation of kiwi allergy was carried out. RESULTS: Act d 11 was identified as a member of the major latex protein/ripening-related protein (MLP/RRP) family. IgE binding to Act d 11 was shown by all the applied testing. Patients tested positive for Act d 11 and reporting symptoms on kiwifruit exposure were found within the Bet v 1-positive subset rather than within the population selected for highly reliable history of allergic reactions to kiwifruit. Epidemiology of Act d 11 IgE reactivity was documented in the two cohorts. IgE co-recognition of Act d 11 within the Bet v 1-like molecules is documented using the microarray IgE inhibition assay. CONCLUSIONS: Act d 11 is the first member of the MLP/RRP protein family to be described as an allergen. It displays IgE co-recognition with allergens belonging to the PR-10 family, including Bet v 1.