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A convenient and sensitive allergy test: IgE crosslinking-induced luciferase expression in cultured mast cells.


ABSTRACT:

Background

For the detection of allergen-specific IgE in sera, solid-phase IgE-binding assays like the CAP test are commonly used. Although such immunochemical methods are very sensitive, they frequently produce false positives. Degranulation of the human IgE receptor (Fc?RI)-transfected rat mast cell (RBL) lines seems to be a possible indicator for human IgE, but spontaneous mediator release from these cells in the presence of human sera is not negligible.

Methods

The nuclear factor of activated T-cells (NFAT)-responsive luciferase reporter gene was stably transfected into human Fc?RI-expressing RBL-SX38 cells. One established clone (RS-ATL8) was sensitized with 1?:?100 dilution of sera from patients with egg white allergy and then stimulated with purified or a crude extract of egg white allergen.

Results

Sensitization with 15?pg/ml IgE was sufficient to detect IgE crosslinking-induced luciferase expression (EXiLE) by anti-IgE stimulation. Allergen-specific EXiLE was elicited by as little as 1?fg/ml of egg white protein without cytotoxicity. There was a good correlation between results with EXiLE and oral food challenge tests on patients with egg allergy (P?=?0.001687, Fisher's exact test). The measured values of EXiLE and the CAP test also correlated well (R?=?0.9127, Spearman's test).

Conclusion

The EXiLE test using RS-ATL8 cells is a promising in vitro IgE test to evaluate the biological activity of the binding between IgE and allergens.

SUBMITTER: Nakamura R 

PROVIDER: S-EPMC3066406 | BioStudies | 2010-01-01

REPOSITORIES: biostudies