Analysis Of Allergens By The Cell Model Of Releasing Histamine In Vitro

AIM:To analysis the irritability of different allergens in food,we established a cell model of releasing histamine by sensitized mast cells in vitro,which was induced to release histamine by different components of allergens in the ELISA plate hole.This method can be used to analysis the strength allergens and to determine the necessary components of allergen antibody chip diagnosis and to evaluate effective of allergen vaccines.METHODS:The mice were sensitized by shrimp and dust mite.Peritoneal mast cells(PMC) were collected and then were induced to release histamine with various kinds of allergens. Histamine was detected using fluorescence and histamine EILSA.RESULTS:Three proteins,80KD,36KD and 21KD,were purified from shrimp allergens. Sensitized PMC were inducted by shrimp allergens to release histamine.The 80KD and 36KD protein had the strongest sensitization effect,then 21KD the weakest.The 80KD and 36KD protein plays a pivotal role in shrimp-induced sensitization.The mixture of shrimp allergens can induce stronger histamine release from the mast cells than shrimp allergen alone,implying that there may be some synergistically effects among those different allergens.The Der f2 protein had stronger sensitization effect than the Der f1 protein,therefore the Der f2 protein plays a pivotal role in dust mite-induced sensitization.Along with Der f2 increased the proportion of histamine release also increased.Fluorescence measured the volume of in vitro release of histamine,sensitive,stable,real and effective.CONCLUSION:The cell model of releasing histamine in vitro can objectively reflect the ability of the different components of shrimp allergens and dust mite allergens by releasing histamine. Detecting histamine release is better than detecting IgE or other material in the serum;compared the model of allergen detection in vivo,the cell model of releasing histamine in vitro is more feasible,operable,and replicability.The model shows similar clinical allergy reaction with those in human basophilic leukocyte histamine assay,thus provides a new colitis model for further studies on food allergy and food allergens analysis.