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碳水化合物对蜂毒过敏原磷脂酶A2参与小鼠体内抗原特异性IgE合成的影响。

Involvement of carbohydrate on phospholipase A2, a bee-venom allergen, in in vivo antigen-specific IgE synthesis in mice.

作者信息

Okano M, Nishizaki K, Satoskar A R, Yoshino T, Masuda Y, Harn D A

机构信息

Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, MA, USA.

出版信息

Allergy. 1999 Aug;54(8):811-8.

Abstract

BACKGROUND

Carbohydrates on allergens are known to be important for allergenicity. However, most findings have been made with epitope analysis. In this study, we investigated the involvement of N-glycan on phospholipase A2 (PLA2), the major allergen of honeybee venom, in in vivo synthesis of specific IgE in mice.

METHODS

CBA/J and C57BL/6 mice were sensitized intranasally with either native or deglycosylated PLA2 in the absence of adjuvant. After repeated sensitization, serum Ab titers against PLA2 were determined. PLA2 was deglycosylated chemically with anhydrous trifluoromethanesulfonic acid (TFMS).

RESULTS

CBA/J mice showed PLA2-specific IgE production after repeated sensitization with native PLA2. They also produced PLA2-specific IgG1 predominantly, suggesting that Th2-type Ab production was induced. When we used deglycosylated PLA2 as a competitor in ELISA for detecting PLA2-specific IgE, deglycosylated PLA2 completely inhibited the binding between native PLA2 and IgE. Deglycosylated PLA2 had the same potential for inducing specific IgE synthesis as native PLA2, since sensitization with deglycosylated PLA2 also elicited IgE production in CBA/J mice.

CONCLUSIONS

These results suggest that carbohydrate on PLA2 is less important than previously thought not only as a dominant IgE epitope but also in synthesis of PLA2-specific IgE in vivo.

摘要

背景

已知变应原上的碳水化合物对变应原性很重要。然而,大多数研究结果是通过表位分析得出的。在本研究中,我们调查了蜜蜂毒液主要变应原磷脂酶A2(PLA2)上的N-聚糖在小鼠体内特异性IgE合成中的作用。

方法

在无佐剂的情况下,用天然或去糖基化的PLA2经鼻致敏CBA/J和C57BL/6小鼠。反复致敏后,测定抗PLA2的血清抗体滴度。用无水三氟甲磺酸(TFMS)对PLA2进行化学去糖基化。

结果

CBA/J小鼠经天然PLA2反复致敏后产生了PLA2特异性IgE。它们还主要产生PLA2特异性IgG1,提示诱导了Th2型抗体产生。当我们在ELISA中使用去糖基化的PLA2作为竞争者检测PLA2特异性IgE时,去糖基化的PLA2完全抑制了天然PLA2与IgE之间的结合。去糖基化的PLA2与天然PLA2诱导特异性IgE合成的潜力相同,因为用去糖基化的PLA2致敏也能在CBA/J小鼠中引发IgE产生。

结论

这些结果表明,PLA2上的碳水化合物不仅作为主要的IgE表位,而且在体内PLA2特异性IgE的合成中,其重要性都比以前认为的要低。

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