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

被引:67
作者
Nakamura, R. [1 ]
Uchida, Y.
Higuchi, M.
Nakamura, R. [1 ]
Tsuge, I. [2 ]
Urisu, A. [2 ]
Teshima, R.
机构
[1] Natl Inst Hlth Sci, Div Novel Foods & Immunochem, Setagaya Ku, Tokyo 1588501, Japan
[2] Fujita Hlth Univ, Dept Pediat, Sch Med, Aichi, Japan
关键词
allergy test; food allergy; IgE; luciferase; mast cell; FC-EPSILON-RI; CARBOHYDRATE DETERMINANTS; HISTAMINE-RELEASE; TRANSFECTED CELLS; SERUM IGE; PATIENT; LINE; ACTIVATION; RECEPTOR; ANTIGEN;
D O I
10.1111/j.1398-9995.2010.02363.x
中图分类号
R392 [医学免疫学];
学科分类号
100102 ;
摘要
P>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 epsilon 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 epsilon 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.
引用
收藏
页码:1266 / 1273
页数:8
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