Heymann P W, Chapman M D, Aalberse R C, Fox J W, Platts-Mills T A
Department of Internal Medicine, University of Virginia Medical Center, Charlottesville 22908.
J Allergy Clin Immunol. 1989 Jun;83(6):1055-67. doi: 10.1016/0091-6749(89)90447-8.
Monoclonal antibody affinity chromatography was used to purify two homologous mite allergens, Der f II from Dermatophagoides farinae and Der p II from D. pteronyssinus. They have the same molecular weight (MW) (15 kd) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, they have similar amino acid compositions, and their N-terminal amino acid sequences differ in only four of the first 35 residues. An excellent correlation was observed between IgE antibody to Der f II and Der p II measured in sera from 65 mite-allergic patients (r = 0.94; p less than 0.001) and between quantitative intradermal skin tests to both allergens. A third allergen (Der f III, MW 29 kd) was purified from D. farinae by repeated gel filtration. In sera from 51 mite-allergic patients, IgE antibody to Der f II, Der f III, and previously purified Der f I (MW 24 kd) was detected in 92%, 16%, and 78% of the sera by radioimmunoassay, respectively. Most patients, 41/51 (80%), demonstrated IgE antibody to more than one allergen. With monoclonal antibodies fully cross-reactive with Der f II and Der p II, a two-site immunoassay was developed for measuring absolute quantities (nanograms or micrograms) of these allergens. In extracts rich in mite-fecal material (n = 5), Der f I and Der p I (group I allergens) and Der f II and Der p II (group II allergens) were measured in ratios of 11:1 to 35:1. Lower ratios (1.1:1 to 7:1) were observed in mite body extracts (n = 6). These experiments clearly define a second group of major dust mite allergens that demonstrate extensive structural and antigenic homology.
采用单克隆抗体亲和层析法纯化了两种同源的螨过敏原，即来自粉尘螨的Der f II和来自户尘螨的Der p II。在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳上，它们具有相同的分子量（MW）（15kd），氨基酸组成相似，并且它们的N端氨基酸序列在前35个残基中仅有4个不同。在65例螨过敏患者的血清中检测到的针对Der f II和Der p II的IgE抗体之间（r = 0.94；p <0.001）以及针对这两种过敏原的定量皮内皮肤试验之间观察到极好的相关性。通过反复凝胶过滤从粉尘螨中纯化出第三种过敏原（Der f III，MW 29kd）。在51例螨过敏患者的血清中，通过放射免疫测定法分别在92％、16％和78％的血清中检测到针对Der f II、Der f III和先前纯化的Der f I（MW 24kd）的IgE抗体。大多数患者，即51例中的41例（80％），表现出针对一种以上过敏原的IgE抗体。利用与Der f II和Der p II完全交叉反应的单克隆抗体，开发了一种双位点免疫测定法来测量这些过敏原的绝对量（纳克或微克）。在富含螨粪便物质的提取物（n = 5）中，Der f I和Der p I（I组过敏原）以及Der f II和Der p II（II组过敏原）的测量比例为11:1至35:1。在螨体提取物（n = 6）中观察到较低的比例（1.1:1至7:1）。这些实验清楚地定义了第二组主要的尘螨过敏原，它们表现出广泛的结构和抗原同源性。
