牛牛天天人人综合影院 士锋生物咨询：大胡蜂毒素引起速发型过敏反应机制

翅目昆虫的叮咬是引起速发型过敏反应的三大主要诱因之一。能引起速发型过敏反应的常见昆虫有蜜蜂、胡蜂、牛虻、蚊子等。大胡蜂作为一种个体较大，毒素富含过敏原，常群起攻击人的昆虫，其危害尤为严重。大胡蜂叮咬后的常见症状有红肿、灼痛、颤抖、支气管收缩、甚至过敏性休克危及生命。胡蜂（vespids）种类多达24种，由于各种条件的限制，目前人们仅对其中一种胡蜂（Vespa crabro）毒素的过敏原进行了初步的研究。

为了进一步深入了解胡蜂毒素过敏原组成成分，中国昆明动物研究所赖仞研究员的团队在对牛虻过敏原分离纯化鉴定的经验基础之上，通过整合色谱层析和免疫杂交等技术发展了一套有效分离纯化识别胡蜂毒素过敏原的方法。应用该方法，我们从大胡蜂（vespa magnifica）毒素中分离纯化到两种新的过敏原Vesp ma5和Vespa ma2，它们分别是25-KDa的抗原5蛋白和35-KDa的透明质酸酶（Hyaluronidase）。IgE immunoblot检测显示：Vesp ma5和Vespa ma2对蜂中毒过敏病人血清的IgE的阳性反应率为91.0%和93.9%，对牛虻过敏病人血清IgE阳性反应率也高达86.5%和91.8%。通过Elisa inhibition检测显示：Vesp ma5和Vespa ma2对蜂毒粗样与蜂中毒过敏病人血清IgE结合的zui大抑制率分别为39.5%和39.4%，对牛虻唾液腺粗样与牛虻过敏病人血清IgE结合的zui大抑制率分别为25.3%和23.8%。刺检测显示：Vesp ma5和Vespa ma2对蜂中毒过敏病人的阳性反应率分别为73%和80%。以上结果从不同方面证明了Vesp ma5和Vespa ma2为大胡蜂（vespa magnifica）毒素中两类主要过敏原，同时也为胡蜂牛虻综合征从另一角度提供了有力的证据。

Purification and Characterization of Two New Allergens from the Venom of Vespa magnifica

Su An1,2,5#, Lingling Chen4#, Ji-Fu Wei3#, Xuening Yang1,5#, Dongying Ma1, Xuemei Xu4, Xueqing Xu1, Shaoheng He3*, Jia Lu1*, Ren Lai1*

Due to poor diagnostic facilities and a lack of medical alertness, allergy to Vespa wasps may be underestimated. Few allergens have been identified from Vespa wasps. Possible native allergen proteins were purified from the wasp venoms （WV） （Vespa magnifica Smith） by gel filtration, ion exchange chromatography, respectively. Their sequences were determined by Edman degradation and cDNA cloning. Their allergenicities were assayed by enzyme-linked immunosorbent assay inhibition tests （ELISA-IT）, immunoblots, and skin prick tests （SPTs）. Their cross allergencities with Tab y 2 and Tab y 5 purified from the horsefly （Tabanus yao Macquart） were also determined. Two native allergens were identified from the WV, respectively. They are a 25-KDa antigen 5 protein （Ag5） （Vesp ma 5） and a 35-KDa hyaluronidase （Vesp ma 2）. They represented major allergens in Vespa magnifica by immunoblots and SPTs. ELISA inhibition of pooled sera IgE reactivity to both the WV and the horsefly salivary gland extracts （HSGE） using four purified allergens （Vesp ma 2, Vesp ma 5 and previously purified Tab y 2 and Tab y 5） was significant. Their cross allergenicities were confirmed by ELISA-IT, immunoblots, and SPTs. They represented the cross reactive allergens from wasp and horsefly and proved the so called wasp-horsefly syndrome.