RESUMEN
To identify the key amino acids (AAs) affecting the allergenicity of hemocyanin (HC) allergens from Chinese mitten crabs, in this study, two epitopes, P1-SHFTGSKSNPEQR and P2-LSPGANTITR were employed and four potential key AAs (P1: F3 and N9 and P2: N6 and R10) were predicted. Mast cell and mouse models revealed that four mutants induced lower levels of immunoglobulin E (IgE) and Th2 type cytokines (15.47-49.89 %), proving that F3, N9, N6, and R10 were the key AAs of two epitopes. Mutants reduce allergic responses via the Th2 pathway. However, the roles of every key AA affecting allergenicity were different (P1-F3 > N9 and P2-N6 > R10). In addition, lower transport and higher efflux were observed in the mutants during transport absorption by Caco-2 cells. The allergenicity of HC was stronger when the transport absorption efficiency of epitopes and mutants was higher and their efflux was lower. Our study provides a novel method for revealing the allergenic molecular mechanisms of food allergens.
Asunto(s)
Alérgenos , Aminoácidos , Hemocianinas , Mastocitos , Animales , Alérgenos/inmunología , Alérgenos/genética , Alérgenos/química , Humanos , Hemocianinas/inmunología , Hemocianinas/química , Ratones , Aminoácidos/inmunología , Células CACO-2 , Mastocitos/inmunología , Mastocitos/metabolismo , Inmunoglobulina E/inmunología , Epítopos/inmunología , Braquiuros/inmunología , Citocinas/metabolismo , Secuencia de Aminoácidos , MutaciónRESUMEN
Next-generation live vaccines are created by autonomous production of nitrated antigens.