Furrie E, Turner M W, Strobel S
Division of Cell and Molecular Biology, Institute of Child Health, London, UK.
Immunology. 1995 Nov;86(3):480-6.
Serum collected 60 min after feeding ovalbumin (OVA) to BALB/c mice transfers specific tolerance to naive recipients via an unknown mechanism. We have now identified a large fragment of the OVA molecule as the putative active moiety. Specific absorption of immunoreactive OVA from tolerogenic serum by immunoaffinity chromatography removed the tolerogenic activity; following elution of the bound OVA and subsequent injection into naive recipients it was possible to demonstrate tolerogenic activity equivalent to that seen with unmanipulated serum. Passage of OVA tolerogenic serum through a bovine serum albumin (BSA)-specific affinity column had no effect on in vivo OVA tolerogenic activity. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using anti-OVA antibodies demonstrated the presence of two bands with apparent molecular weights (MW) of 21,000 and 24,000 in the OVA-related material absorbed from the tolerogenic serum. The 24,000 MW moiety was also visualized by silver staining. These moieties are putative candidate tolerogens as they were absent from normal BALB/c mouse serum analysed 5 min after feeding OVA and from severe combined immunodeficiency (SCID) mouse serum--without tolerising activity--analysed 60 min after a similar feed.
给BALB/c小鼠喂食卵清蛋白（OVA）60分钟后收集的血清通过未知机制将特异性耐受性传递给未接触过抗原的受体。我们现已确定OVA分子的一个大片段为假定的活性部分。通过免疫亲和色谱从致耐受性血清中特异性吸收免疫反应性OVA可去除致耐受性活性；洗脱结合的OVA并随后注射到未接触过抗原的受体中后，可证明其致耐受性活性与未处理血清所见相当。OVA致耐受性血清通过牛血清白蛋白（BSA）特异性亲和柱对体内OVA致耐受性活性没有影响。使用抗OVA抗体进行的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹显示，从致耐受性血清中吸收的OVA相关物质中存在两条带，表观分子量（MW）分别为21,000和24,000。24,000 MW的部分也通过银染可视化。这些部分是假定的候选耐受原，因为在喂食OVA 5分钟后分析的正常BALB/c小鼠血清以及在类似喂食60分钟后分析的无致耐受性活性的严重联合免疫缺陷（SCID）小鼠血清中均不存在。
