Usawattanakul W, Tapchaisri P, Thammapalerd N, Tharavanij S, Kojima S
J Parasitol. 1982 Jun;68(3):398-401.
Attempts were made to demonstrate the presence of allergenic components in an extract of axenically cultured Entamoeba histolytica (HK 9). Various strains of mice were immunized i.p. with a low dose of the extract mixed with Al(OH)3 and boosted with the same mixture 4 wk later. A high titer of IgE antibody response was demonstrated in BALB/c, DBA/2 (H-2d), CBA (H-2k), and SWM strains but not in C3H/He (H-2k) strain. The extract was fractionated by gel filtration with Sephadex G-200 and then by DEAE-cellulose chromatography with a gradient buffer with progressive increase in molarity. Fractions having the strongest activity to stimulate the IgE antibody response in the mouse were subjected to SDS polyacrylamide gel electrophoresis. From the results of gel filtration and polyacrylamide gel electrophoresis, the molecular weight on the partially purified allergen was estimated to be 25,000 to 27,000 daltons.
人们试图证明在无菌培养的溶组织内阿米巴(HK 9)提取物中存在变应原成分。用低剂量提取物与氢氧化铝混合腹腔注射免疫各种品系小鼠,并在4周后用相同混合物加强免疫。在BALB/c、DBA/2(H-2d)、CBA(H-2k)和SWM品系小鼠中显示出高滴度的IgE抗体反应,但在C3H/He(H-2k)品系小鼠中未显示。提取物先用葡聚糖G-200进行凝胶过滤,然后用梯度缓冲液进行DEAE-纤维素柱层析,摩尔浓度逐渐增加。对刺激小鼠IgE抗体反应活性最强的级分进行SDS聚丙烯酰胺凝胶电泳。根据凝胶过滤和聚丙烯酰胺凝胶电泳结果,部分纯化变应原的分子量估计为25000至27000道尔顿。