ANTIGENIC AND STRUCTURAL-ANALYSIS OF GROUP-II ALLERGENS (DER-F-II AND DER-P-II) FROM HOUSE DUST MITES (DERMATOPHAGOIDES SPP)
ANTIGENIC AND STRUCTURAL-ANALYSIS OF GROUP-II ALLERGENS (DER-F-II AND DER-P-II) FROM HOUSE DUST MITES (DERMATOPHAGOIDES SPP)
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DOI：
10.1016/0091-6749(89)90447-8
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发表时间：
1989-06-01
影响因子：
14.2
通讯作者：
PLATTSMILLS, TAE
中科院分区：
文献类型：
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作者：
HEYMANN, PW;CHAPMAN, MD;PLATTSMILLS, TAE
使用单克隆亲和层析纯化两种同源螨过敏原：来自 Dermatophagoides ferinae 的 Der f II 和来自 D. pteronyssinus 的 Der p II。它们在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上具有相同的分子量（MW）（15 kd），它们具有相似的氨基酸组成，并且它们的N端氨基酸序列仅在前35个残基中的4个不同。在 65 名螨过敏患者的血清中测量的 Der f II 和 Der p II 的 IgE 抗体之间以及对两种过敏原的定量皮内皮肤测试之间观察到极好的相关性（r = 0.94；p < 0.001）。通过重复凝胶过滤从 D. farinae 中纯化出第三种过敏原（Der f III，MW 29 kd）。在 51 英里过敏患者的血清中，通过放射免疫测定法分别检测到 92%、16% 和 78% 的血清中的 Der f II、Der f III 和先前纯化的 Der f I (MW 24 kd) 的 IgE 抗体。大多数患者（41/51 (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）。在富含螨虫粪便物质的提取物中 (n = 5)，以 11:1 至 35:1 的比例测量 Der f I 和 Der p I（I 类过敏原）以及 Der f II 和 Der p II（II 类过敏原）。在螨体提取物中观察到较低的比率（1.1:1 至 7:1）（n = 6）。这些实验清楚地定义了第二组主要尘螨过敏原，它们表现出广泛的结构和抗原同源性。
Monoclonal affinity chromatography was used to purify two homologous mite allergens, Der f II from Dermatophagoides ferinae 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 < 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 mile-allergic patients, IgE antibody to Der f II, Der f III, and previously purified Der f I ( MW 24 kd) was detected 92%, 16%, and 78% of the sera by radioimmmunoassay, 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.