Allergy Department, Hospital Universitari Germans Trias i Pujol, Universitat Autonoma de Barcelona, Badalona, Spain.
Int Arch Allergy Immunol. 2012;159(2):143-6. doi: 10.1159/000336388. Epub 2012 May 30.
The existence of IgE binding to dog dander extract without IgE antibodies against the described dog allergens (Can f 1, 2, 3 and 4) implies the presence of other dog allergens yet to be identified. Recently, an IgE-binding protein was isolated from dog urine and identified as prostatic kallikrein; it has been named Can f 5. Cross-reactivity between a dog dander allergen and human prostate-specific antigen (PSA) has been described. The aim of this study was to identify the dog dander allergen that presents cross-reactivity with PSA and demonstrate its clinical relevance in our patient with human seminal plasma allergy.
SDS-PAGE immunoblotting and inhibition tests were performed. Mass spectrometry was carried out to identify the protein involved in the allergy reactions.
SDS-PAGE immunoblotting-inhibition with an IgE-binding protein from dog prostatic secretion showed total IgE binding inhibition to a 28-kDa IgE-reactive band identified as PSA. The electroeluted protein from dog prostatic secretion was identified by mass spectrometry as Can f 5. IgE immunoblotting of human seminal plasma incubated with the serum of the patient revealed two IgE-binding bands (28 and 32.7 kDa). Both SDS-PAGE immunoblotting inhibition assays, with human seminal plasma or purified PSA in solid phase, showed complete IgE binding inhibition when the serum of the anaphylactic patient was preincubated with dog dander extract or recombinant Can f 5.
The dog dander allergen that shows cross-reactivity with human PSA has been characterized and turns out to be the recently described Can f 5. We demonstrated the clinical relevance of this cross-reactivity in a patient.
存在针对狗皮屑提取物的 IgE 结合，而没有针对所述狗过敏原（Can f 1、2、3 和 4）的 IgE 抗体，这意味着存在其他尚未鉴定的狗过敏原。最近，从狗尿中分离出一种 IgE 结合蛋白，并将其鉴定为前列腺激肽；它被命名为 Can f 5。已描述了狗皮屑过敏原与人前列腺特异性抗原（PSA）之间的交叉反应。本研究的目的是鉴定与 PSA 呈现交叉反应的狗皮屑过敏原，并在我们患有人类精液过敏症的患者中证明其临床相关性。
进行 SDS-PAGE 免疫印迹和抑制试验。进行质谱分析以鉴定参与过敏反应的蛋白质。
用来自狗前列腺分泌物的 IgE 结合蛋白进行 SDS-PAGE 免疫印迹抑制试验，显示对 28 kDa IgE 反应带的总 IgE 结合抑制，该带被鉴定为 PSA。从狗前列腺分泌物中电洗脱的蛋白质通过质谱鉴定为 Can f 5。用人精液与患者血清孵育的 IgE 免疫印迹显示两条 IgE 结合带（28 和 32.7 kDa）。用固相中的人精液或纯化的 PSA 进行的两种 SDS-PAGE 免疫印迹抑制试验均显示，当过敏患者的血清与狗皮屑提取物或重组 Can f 5 预孵育时，完全抑制了 IgE 结合。
与人类 PSA 发生交叉反应的狗皮屑过敏原已被鉴定，事实证明它是最近描述的 Can f 5。我们在患者中证明了这种交叉反应的临床相关性。
