近日,,國(guó)際著名雜志PLoS One在線刊登了中國(guó)科學(xué)院昆明動(dòng)物研究所研究人員的最新研究成果“Purification and Characterization of Two New Allergens from the Venom of Vespa magnifica,,”,,文章中,研究者在大胡蜂毒素引起速發(fā)型過(guò)敏反應(yīng)研究取得重要的進(jìn)展,。
膜翅目昆蟲(chóng)的叮咬是引起速發(fā)型過(guò)敏反應(yīng)的三大主要誘因之一,。能引起速發(fā)型過(guò)敏反應(yīng)的常見(jiàn)昆蟲(chóng)有蜜蜂、胡蜂,、牛虻,、蚊子等。大胡蜂作為一種個(gè)體較大,,毒素富含過(guò)敏原,,常群起攻擊人的昆蟲(chóng),其危害尤為嚴(yán)重,。大胡蜂叮咬后的常見(jiàn)癥狀有紅腫,、灼痛,、顫抖、支氣管收縮,、甚至過(guò)敏性休克危及生命,。胡蜂(vespids)種類多達(dá)24種,由于各種條件的限制,,目前人們僅對(duì)其中一種胡蜂(Vespa crabro)毒素的過(guò)敏原進(jìn)行了初步的研究,。
為了進(jìn)一步深入了解胡蜂毒素過(guò)敏原組成成分,中國(guó)科學(xué)院昆明動(dòng)物研究所賴仞研究員領(lǐng)導(dǎo)的團(tuán)隊(duì)在對(duì)牛虻過(guò)敏原分離純化鑒定的經(jīng)驗(yàn)基礎(chǔ)之上,,通過(guò)整合色譜層析和免疫雜交等技術(shù)發(fā)展了一套有效分離純化識(shí)別胡蜂毒素過(guò)敏原的方法。應(yīng)用該方法,,我們從大胡蜂(vespa magnifica)毒素中分離純化到兩種新的過(guò)敏原Vesp ma5和Vespa ma2,,它們分別是25-KDa的抗原5蛋白和35-KDa的透明質(zhì)酸酶(Hyaluronidase)。IgE immunoblot檢測(cè)顯示:Vesp ma5和Vespa ma2對(duì)蜂中毒過(guò)敏病人血清的IgE的陽(yáng)性反應(yīng)率為91.0%和93.9%,,對(duì)牛虻過(guò)敏病人血清IgE陽(yáng)性反應(yīng)率也高達(dá)86.5%和91.8%,。通過(guò)Elisa inhibition檢測(cè)顯示:Vesp ma5和Vespa ma2對(duì)蜂毒粗樣與蜂中毒過(guò)敏病人血清IgE結(jié)合的最大抑制率分別為39.5%和39.4%,對(duì)牛虻唾液腺粗樣與牛虻過(guò)敏病人血清IgE結(jié)合的最大抑制率分別為25.3%和23.8%,。皮膚針刺檢測(cè)顯示:Vesp ma5和Vespa ma2對(duì)蜂中毒過(guò)敏病人的陽(yáng)性反應(yīng)率分別為73%和80%,。以上結(jié)果從不同方面證明了Vesp ma5和Vespa ma2為大胡蜂(vespa magnifica)毒素中兩類主要過(guò)敏原,同時(shí)也為胡蜂牛虻綜合征從另一角度提供了有力的證據(jù),。
該成果加深了我們對(duì)大胡蜂毒素過(guò)敏原成分的了解,,對(duì)大胡蜂及其它昆蟲(chóng)刺蟄引起過(guò)敏反應(yīng)的防治具有重要意義。該項(xiàng)目研究得到了國(guó)家自然科學(xué)基金的經(jīng)費(fèi)資助,。(生物谷Bioon.com)
doi:10.1371/journal.pone.0031920
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PMID:
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.