Romagnani S, Boccaccini P, Amadori A, Ricci M
Int Arch Allergy Appl Immunol. 1976;50(5):525-35. doi: 10.1159/000231557.
Dermatophagoides pteronyssinus (DP) extract was fractionated by Sephadex G-75 and liquid isoelectric focusing (IEF) and the allergenic activity of different fractions was monitored by direct and indirect RAST. The fractionation on Sephadex G-75 showed that the allergenic activity of DP extract was related to wide molecular weight spectrum components, even though the maximum amount was recovered in effluent that contained protein with a molecular weight ranging between 25,000 and 12,500 daltons. By fractionation of the mite extract on IEF, three main peaks of allergenic activity (pI less than 3.0; pI = 4.3 +/- 0.25; pI = 6.4 +/- 0.25) were found. Cross-inhibition experiments showed a high degree of cross-reactivity between allergenic material eluted in very distant regions of molecular weight or isoelectric point. The allergenic activity of unfractionated mite extract and of its IEF fractions was destroyed by pronase - but not by neuraminidase - treatment. These results suggest that DP extract probably contains one main allergen existing in multiple molecular forms rather than several distinct allergens and that a protein moiety of the allergen is necessary for the combination with IgE.
用葡聚糖G - 75和液相等电聚焦（IEF）对粉尘螨提取物进行分级分离，并通过直接和间接放射变应原吸附试验（RAST）监测不同级分的变应原活性。葡聚糖G - 75分级分离结果表明，粉尘螨提取物的变应原活性与较宽分子量范围的组分有关，尽管在含有分子量介于25,000至12,500道尔顿之间蛋白质的流出物中回收量最大。通过对螨提取物进行IEF分级分离，发现了三个主要的变应原活性峰（等电点小于3.0；等电点 = 4.3 ± 0.25；等电点 = 6.4 ± 0.25）。交叉抑制实验表明，在分子量或等电点非常不同区域洗脱的变应原物质之间存在高度交叉反应性。未分级的螨提取物及其IEF级分的变应原活性可被链霉蛋白酶破坏，但不能被神经氨酸酶破坏。这些结果表明，粉尘螨提取物可能含有一种以多种分子形式存在的主要变应原，而非几种不同的变应原，并且变应原的蛋白质部分对于与IgE结合是必需的。
