Supplementary Material for: Arginine Kinase from the Cellar Spider <i>(Holocnemus pluchei): </i>A New Asthma-Causing Allergen
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<i>Background:</i> We report a 31-year-old farmer whose work consists in handling cereal and vegetables, who consulted our clinic because of asthma symptoms after inhalation of dust during manipulation of the deposited material, usually inside the warehouse. <i>Methods and Results:</i> Skin prick tests and specific immunoglobulin E (IgE) determinations were negative with common aeroallergens. The patient noted the presence of many spiders in the warehouse, which were identified as the cellar spider <i>Holocnemus pluchei </i>and the common house spider<i>, Tegenaria domestica. </i>Extracts of spider bodies brought in by the patient were obtained and used to perform in vivo and in vitro studies. Molecular characterization of IgE-binding bands was performed by mass spectrometry. We obtained positive prick tests to the extracts of the bodies of both spiders. Immunoblotting displayed different bands in both spider extracts, in a range of 20–70 kDa. All were hemocyanins, except for a 17-kDa protein of <i>Holocnemus</i> identified as an arginine kinase (AK). Bronchial challenge was positive with the extract of the cellar spider and with the AK, but was negative with the domestic house spider. <i>Conclusion:</i> We present the first case of respiratory allergy due to sensitization to AK from a common spider, confirmed by bronchial provocation tests.
**背景**：本研究报道1例31岁谷物与蔬菜装卸工人病例，该患者因在仓库内操作堆放物料时吸入粉尘后出现哮喘症状，前往我院门诊就诊。 **方法与结果**：针对常见气源性变应原的皮肤点刺试验与特异性免疫球蛋白E（IgE）检测均为阴性。患者自述仓库内存在大量蜘蛛，经鉴定分别为地窖蜘蛛（Holocnemus pluchei）与家隅蛛（Tegenaria domestica）。采集患者携带的蜘蛛躯体提取物，开展体内与体外实验研究。通过质谱技术对IgE结合条带进行分子表征。两种蜘蛛躯体提取物的皮肤点刺试验均呈阳性。免疫印迹结果显示，两种蜘蛛提取物中均存在分子量20~70 kDa的差异化条带，其中除地窖蜘蛛的17 kDa蛋白被鉴定为精氨酸激酶（AK）外，其余均为血蓝蛋白。地窖蜘蛛提取物与精氨酸激酶的支气管激发试验均为阳性，而家隅蛛提取物的支气管激发试验结果为阴性。 **结论**：本研究报道首例因接触常见蜘蛛精氨酸激酶致敏引发呼吸道过敏的病例，该病例经支气管激发试验证实。
