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利用CD63上调和IgE下调进行变应原诱导的嗜碱性粒细胞活化的流式细胞术分析。活化指数的定义。

Use of both CD63 up regulation and IgE down regulation for the flow cytometric analysis of allergen induced basophil activation. Definition of an activation index.

作者信息

Sainte-Laudy J, Boumediene A, Touraine F, Orsel I, Brianchon C, Bonnaud F, Cogné M

机构信息

Laboratoire d'Immunologie, Hôpital universitaire Dupuytren, 2, avenue Martin-Luther-King, 87042 Limoges Cedex, France.

出版信息

Inflamm Res. 2007 Jul;56(7):291-6. doi: 10.1007/s00011-007-7014-5.

Abstract

OBJECTIVE AND DESIGN

The aim of this study was to compare the use of a late (CD63) and an early (IgE) marker of basophil activation in the flow cytometric diagnosis of beta-lactam induced allergic hypersensitivity reactions.

SUBJECTS

Twelve patients who had had a clear cut betalactam induced immediate reaction and 16 controls were selected, as well as 11 patients who had had an immediate reaction to bee or wasp stings.

METHODS

Leukocyte suspensions were incubated with allergen dilutions as well as 2 positive controls (anti-IgE and NFormyl- Methionyl-Leucyl-Phenylalanine (fMLP)). Basophils were labelled with an anti-IgE FITC (fluorescein isothiocyanate) and an anti-CD63 PE (phycoerythrin). Results were expressed as percentage CD63 expression and index calculated according to a specific algorithm including the two activation markers.

RESULTS

Significant CD63 expression (>5 %) was observed in 3/12 cases for the beta-lactam sensitized population, in 0/16 cases for the controls and in 11/11 cases for the venom sensitized population. A significant index (determined by a ROC analysis) was observed in 11/12 beta-lactam sensitized patients and in 0/16 controls.

CONCLUSION

These results show that IgE (an early activation marker) is more sensitive than CD63 (a later activation marker) in the diagnosis of beta-lactam allergy.

摘要

目的与设计

本研究旨在比较在流式细胞术诊断β-内酰胺诱导的过敏性超敏反应中,嗜碱性粒细胞活化的晚期标志物(CD63)和早期标志物(IgE)的应用情况。

受试者

选取12例有明确的β-内酰胺诱导的速发型反应的患者、16例对照以及11例对蜜蜂或黄蜂蜇伤有速发型反应的患者。

方法

将白细胞悬液与过敏原稀释液以及2种阳性对照(抗IgE和N-甲酰甲硫氨酰亮氨酰苯丙氨酸(fMLP))一起孵育。用抗IgE异硫氰酸荧光素(FITC)和抗CD63藻红蛋白(PE)标记嗜碱性粒细胞。结果以CD63表达百分比表示,并根据包含这两种活化标志物的特定算法计算指数。

结果

在β-内酰胺致敏人群中,12例中有3例观察到显著的CD63表达(>5%),对照的16例中无1例出现,毒液致敏人群的11例中11例出现。在12例β-内酰胺致敏患者中的11例以及16例对照中的0例观察到显著指数(由ROC分析确定)。

结论

这些结果表明,在β-内酰胺过敏诊断中,IgE(早期活化标志物)比CD63(晚期活化标志物)更敏感。

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