Severe IgE-mediated anaphylaxis following consumption of fried frog legs: Definition of α-parvalbumin as the allergen in cause

Background: IgE-mediated allergic reactions to bullfrog and edible frog have been reported. The implicated allergens have not been defined so far. The frog material and the patient's serum from a case of severe food-induced anaphylaxis were used to define the implicated allergen at the protein and DNA level. Methods: Immunoblotting techniques and N-terminal protein microsequencing were used to define the allergen recognized by the patient's serum. Back translation from the identified protein sequence was used to design degenerated primers to amplify the allergen's cDNA by polymerase chain reaction (PCR). We defined the nucleotide sequence of the allergen from the frog of Indonesian origin that was consumed by the patient, and the homologous cDNA from Rana esculenta. Results: Protein microsequencing revealed that the implicated frog allergen belonged to the parvalbumin family. cDNAs coding for α- and β-parvalbumin of R. esculenta and Rana species were cloned. Recombinant proteins were expressed in Escherichia coli. The patient's serum IgE antibodies recognized parvalbumin prepared from frog muscle and recombinant α-parvalbumin from R. species but not from R. esculenta. Recombinant β-parvalbumin was not recognized by the IgE antibodies. Conclusion: This work defines at the protein and DNA levels α-parvalbumin as the allergen implicated in a case of IgE-mediated anaphylaxis to frog muscle. It also shows that a protein belonging to the parvalbumin family is implicated in type I allergies outside the fish species.