Hilger Christiane, Dubey Ved P, Lentz Delphine, Davril Caroline, Revets Dominique, Muller Claude P, Diederich Claire, De La Barrière Hélène, Codreanu-Morel Françoise, Morisset Martine, Lehners Christiane, De Prabir K, Hentges François
Departments of Infection and Immunity, Luxembourg Institute of Health, Luxembourg, Luxembourg.
Int Arch Allergy Immunol. 2015;166(1):30-40. doi: 10.1159/000371420. Epub 2015 Feb 28.
An increasing number of asthma cases upon exposure to hamsters and anaphylactic reactions following hamster bites are being reported, but the allergens responsible are still poorly characterized. In the Golden hamster, male-specific submaxillary gland protein (MSP), a lipocalin expressed in a sex- and tissue-specific manner in the submaxillary and lacrimal glands, is secreted in the saliva, tears and urine. The purpose of this study was to determine if MSP is an allergen, to identify IgE-reactive proteins of different hamster species and to analyse potential cross-reactivities.
Fur extracts were prepared from four hamster species. Hamster-allergic patients were selected based on a history of positive IgE-test to hamster epithelium. The IgE-reactivity of patients' sera was investigated by means of immunoblot and ELISA. IgE-reactive proteins in fur extracts and the submaxillary gland were identified using anti-MSP antibodies, Edman sequencing or mass spectrometry. MSP was purified from Golden hamster and recombinant MSP was expressed in E. coli.
Four patients had IgE-antibodies against 20.5-kDa and 24-kDa proteins of Golden hamster fur extract, which were identified as MSP. IgE-reactive MSP-like proteins were detected in European hamster fur extract. Three patient sera showed IgE-reactive bands at 17-21 kDa in Siberian and Roborovski hamster fur extracts. These proteins were identified as two closely related lipocalins. Immunoblot inhibition experiments showed that they are cross-reactive and are different from MSP.
MSP lipocalin of the Golden hamster was identified as an allergen, and it is different from the cross-reactive lipocalin allergens of Siberian and Roborovski hamsters. Our findings highlight the need for specific tools for the in vitro and in vivo diagnosis of allergy to different hamster species.
据报道,接触仓鼠后哮喘病例增多,仓鼠咬伤后出现过敏反应,但相关过敏原仍未得到充分表征。在金黄仓鼠中,雄性特异性颌下腺蛋白(MSP)是一种在颌下腺和泪腺中以性别和组织特异性方式表达的脂质运载蛋白,存在于唾液、眼泪和尿液中。本研究的目的是确定MSP是否为过敏原,鉴定不同仓鼠物种的IgE反应性蛋白,并分析潜在的交叉反应性。
从四种仓鼠物种制备毛发提取物。根据对仓鼠上皮IgE检测呈阳性的病史选择对仓鼠过敏的患者。通过免疫印迹和ELISA研究患者血清的IgE反应性。使用抗MSP抗体、埃德曼测序或质谱鉴定毛发提取物和颌下腺中的IgE反应性蛋白。从金黄仓鼠中纯化MSP,并在大肠杆菌中表达重组MSP。
四名患者对金黄仓鼠毛发提取物中的20.5 kDa和24 kDa蛋白具有IgE抗体,这些蛋白被鉴定为MSP。在欧洲仓鼠毛发提取物中检测到IgE反应性MSP样蛋白。三名患者血清在西伯利亚仓鼠和罗伯罗夫斯基仓鼠毛发提取物中显示出17 - 21 kDa的IgE反应条带。这些蛋白被鉴定为两种密切相关的脂质运载蛋白。免疫印迹抑制实验表明它们具有交叉反应性,且与MSP不同。
金黄仓鼠的MSP脂质运载蛋白被鉴定为过敏原,它与西伯利亚仓鼠和罗伯罗夫斯基仓鼠具有交叉反应性的脂质运载蛋白过敏原不同。我们的研究结果强调了需要用于体外和体内诊断对不同仓鼠物种过敏的特异性工具。