Ovsyannikova I G, Vailes L D, Li Y, Heymann P W, Chapman M D
UVA Asthma and Allergic Diseases Center, Charlottesville 22908.
J Allergy Clin Immunol. 1994 Sep;94(3 Pt 1):537-46. doi: 10.1016/0091-6749(94)90211-9.
Group II allergens are a major cause of sensitization in patients allergic to mites. To facilitate the antigenic analysis of group II allergens and to develop improved methods of allergen detection, we compared IgG anti-group II antibody responses in inbred mouse strains and raised a panel of monoclonal antibodies (mAbs).
IgE antibody responses were compared by antigen-binding radioimmunoassay. Epitope specificity of the mAbs was analyzed by two-site binding assays and by cross-inhibition radioimmunoassays.
Comparison of polyclonal IgG antibody responses in five BALB congenic strains showed that H-2d mice had poor responses, whereas H-2b and H-2k mice had strong, cross-reactive, IgG anti-group II responses. The specificities of nine anti-Der p II IgE mAbs raised in A/J mice were compared with specificities of seven mAbs produced previously. Most mAbs (11 of 16) recognized common epitopes on Der p II and Der f II: three were specific to Der p II, and two showed high binding to Der f II. Epitope analysis showed that the mAbs defined four cross-reactive, nonoverlapping sites on the group II allergens. Binding of several combinations of mAbs was compared, and a two-site ELISA for group II antigens was developed. Linear regression analysis showed an excellent correlation between results of this assay and group II radioimmunoassay of house dust samples (n = 40, r = 0.85, p < 0.001).
There are multiple cross-reactive B-cell epitopes on group II allergens. The group II ELISA has several important applications, including assessment of environmental allergen exposure, monitoring of the efficacy of avoidance procedures, and standardization of commercial mite allergen extracts.
II类变应原是对螨类过敏患者致敏的主要原因。为便于对II类变应原进行抗原分析并开发改进的变应原检测方法，我们比较了近交系小鼠品系中IgG抗II类抗体反应，并制备了一组单克隆抗体（mAb）。
通过抗原结合放射免疫测定比较IgE抗体反应。通过双位点结合测定和交叉抑制放射免疫测定分析mAb的表位特异性。
对五个BALB同源品系的多克隆IgG抗体反应进行比较，结果显示H-2d小鼠反应较弱，而H-2b和H-2k小鼠具有较强的、交叉反应性的IgG抗II类反应。比较了在A/J小鼠中产生的9种抗Der p II IgE mAb的特异性与先前产生的7种mAb的特异性。大多数mAb（16种中的11种）识别Der p II和Der f II上的共同表位：3种对Der p II具有特异性，2种与Der f II具有高结合力。表位分析表明，这些mAb在II类变应原上定义了4个交叉反应性、不重叠的位点。比较了几种mAb组合的结合情况，并开发了一种用于II类抗原的双位点ELISA。线性回归分析表明，该检测结果与室内灰尘样本的II类放射免疫测定结果之间具有良好的相关性（n = 40，r = 0.85，p < 0.001）。
II类变应原上存在多个交叉反应性B细胞表位。II类ELISA有几个重要应用，包括评估环境变应原暴露、监测回避措施的效果以及商业螨变应原提取物的标准化。
