MONOCLONAL IMMUNOASSAYS FOR MAJOR DUST MITE (DERMATOPHAGOIDES) ALLERGENS, DER-P-I AND DER-F-I, AND QUANTITATIVE-ANALYSIS OF THE ALLERGEN CONTENT OF MITE AND HOUSE DUST EXTRACTS
MONOCLONAL IMMUNOASSAYS FOR MAJOR DUST MITE (DERMATOPHAGOIDES) ALLERGENS, DER-P-I AND DER-F-I, AND QUANTITATIVE-ANALYSIS OF THE ALLERGEN CONTENT OF MITE AND HOUSE DUST EXTRACTS
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DOI：
10.1016/0091-6749(87)90128-x
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发表时间：
1987-08-01
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：
CHAPMAN, MD;HEYMANN, PW;PLATTSMILLS, TAE
建立单克隆双位点放射免疫分析法（RIA），测定屋尘螨变应原Der p I和尘螨变应原Der f I。花粉将包被单克隆抗体（Mab）的微量滴定板与螨提取物一起孵育，并用具有不同表位特异性的第二个125 I标记的Mab检测结合的过敏原。Mab RIA非常敏感（纳克范围）和高度特异性。D.不同浓度的Der p I与屋盖蕨提取物的结合曲线呈平行曲线，而Der p I与屋盖蕨提取物的结合曲线呈平行曲线。在相同的测定中，花粉提取物显示出< 5%的Der p I结合。几种D. farinae提取物在Der f I测定中的活性，而D.屋盖蕨提取物显示很少或没有结合。将Mab RIA与测定Der p I和Der f I交叉反应决定簇（抗原P1当量[AgP 1 Eq]）的抑制RIA进行比较。结果表明，在商业螨和屋尘提取物中，这些测定之间具有良好的定量一致性（平均差1.57 ± 0.001）。0.5倍）。30个已知螨数的房屋灰尘样本。还比较了Der p I和AgP 1 Eq含量。Der p I和Der f I的Mab RIA值总和与AgP 1 Eq值（r = 0.86; p < 0.001）和通过RAST抑制评估的螨变应原总含量（n = 21，r = 0.77，p < 0.001）具有非常好的相关性。此外，在每100 mg灰尘中有超过10只螨的样品中，Der p I：Der f I比值与显微镜计数的两种螨的比值密切相关（n = 15，r = 0.89; p < 0.001）。Mab RIA可以测量螨或灰尘提取物中的过敏原水平，而不需要纯化的过敏原或亲和纯化的抗体，并且可以容易地标准化。这些试验将有助于过敏性哮喘的流行病学研究，以评估患者对螨过敏原的暴露，以及避免方案的影响。由于试剂的长期稳定性和重现性。针对特定过敏原的单克隆抗体检测也将在螨和其他过敏原提取物的标准化中发挥重要作用。
Monoclonal, two-site radioimmunoassay (RIAs) were developed to measure allergen Der p I of Dermatophagoides pteronyssinus or Der f I of D. farinae. Microtiter plates coated with monoclonal antibody (Mab) were incubated with mite extract, and bound allergen was detected with a second 125I-labeled Mab of different epitope specificity. The Mab RIAs were very sensitive (nanogram range) and highly specific. D. pteronyssinus extracts with different concentrations of Der p I demonstrated parallel binding curves, whereas a potent D. farinae extract demonstrated < 5% of the Der p I binding in the same assay. Similar parallel curves were obtained with several D. farinae extracts in the Der f I assay, whereas D. pteronyssinus extract demonstrated little or no binding. The Mab RIAs were compared with an inhibition RIA that measured cross-reacting determinants on both Der p I and Der f I (antigen P1 equivalent [AgP1Eq]). The results demonstrated good quantitative agreement between these assays in commercial mite and house dust extracts (mean difference 1.57 .+-. 0.5-fold). Thirty house dust samples with known mite counts. Der p I, and AgP1Eq content were also compared. The summed Mab RIA values for Der p I and Der f I demonstrated a very good correlations with AgP1Eq values (r = 0.86; p < 0.001) and with assessments of total mite-allergen content by RAST inhibition (n = 21, r = 0.77, p < 0.001). Furthermore, in samples with more than 10 mites per 100 mg of dust, the Der p I: Der f I ratio closely correlated with the ratio of the two mites counted by microscopy (n = 15, r = 0.89; p < 0.001). The Mab RIAs can measure allergen levels in mite or dust extracts without the need for purified allergen or affinity-purified antibodies and can readily be standardized. These assays will be useful in epidemiologic studies of allergic asthma, to assess patients'' exposure to mite allergens, and the effects of avoidance regimens. Because of the long-term stability and reproducibility of the reagents. Mab-based assays for specific allergens will also play an important role in the standardization of mite and other allergens extracts.