Shigeta S, Tanaka S, Yoshizane T, Yamaguchi T, Fujii S, Jyo T, Wada T, Oka S, Ono K
Department of Fermentation Technology, Faculty of Engineering, Hiroshima University.
Arerugi. 1995 Mar;44(3 Pt 1):170-81.
A series of mite allergens with high carbohydrate content was fractionated from mite extract containing mite feces by ultrafiltration, and successive chromatography on Ultrogel AcA 54, Sepharose CL-6B, and DEAE Toyopearl 650S. The final fractions isolated were termed HM1-2-Eff, -0.1A, -0.1B, -0.3A, -0.3B, and -1M according to the eluted salt concentration. These six fractions were stained weakly with Coomassie Brilliant Blue but strongly with periodate-Schiff's reagent on SDS-PAGE. Their molecular weight were estimated to be 150-155 KD on Sepharose CL-6B. All of these six fractions provoked strong erythema in skin of mite-sensitive and asthmatic patients. HM1-2-Eff, -0.1A, -0.1B and -0.3B reacted with 73% of mite-sensitive patients' IgE by the nitrocellulose dot blotting. One ng/ml of 4 fractions (HM1-2-0.1A, 0.3A, 0.3B and -1M) released histamine from mite allergic patient's blood cells. HM1-2-0.3A induced proliferation of T cell from immunized mouse by mite extract containing mite feces (Dff).
通过超滤以及在Ultrogel AcA 54、Sepharose CL - 6B和DEAE Toyopearl 650S上的连续层析，从含有螨粪便的螨提取物中分离出一系列高碳水化合物含量的螨过敏原。根据洗脱盐浓度，最终分离出的级分分别称为HM1 - 2 - Eff、- 0.1A、- 0.1B、- 0.3A、- 0.3B和- 1M。在SDS - PAGE上，这六个级分用考马斯亮蓝染色较弱，但用高碘酸 - 希夫试剂染色较强。在Sepharose CL - 6B上估计它们的分子量为150 - 155 KD。所有这六个级分在螨敏感和哮喘患者的皮肤上都引起了强烈的红斑。通过硝酸纤维素斑点印迹法，HM1 - 2 - Eff、- 0.1A、- 0.1B和- 0.3B与73%的螨敏感患者的IgE发生反应。1 ng/ml的4个级分（HM1 - 2 - 0.1A、0.3A、0.3B和- 1M）从螨过敏患者的血细胞中释放组胺。HM1 - 2 - 0.3A通过含有螨粪便的螨提取物（Dff）诱导免疫小鼠的T细胞增殖。
