Antibody response to aggregated human interferon alpha2b in wild-type and transgenic immune tolerant mice depends on type and level of aggregation
Antibody response to aggregated human interferon alpha2b in wild-type and transgenic immune tolerant mice depends on type and level of aggregation
复制标题
DOI：
10.1002/jps.20599
复制
发表时间：
2006-05-01
影响因子：
3.8
通讯作者：
Jiskoot, W
中科院分区：
文献类型：
--
作者：
Hermeling, S;Schellekens, H;Jiskoot, W
本研究的目的是确定转基因免疫耐受小鼠对重组人干扰素α 2b （rhIFN α 2b）的类型和聚集水平的敏感性。RhIFN α 2b通过金属催化氧化或在高温和不同ph值下孵育而聚集。将天然rhIFNa2b与氧化后的rhIFNa2b按不同比例混合，得到不同聚集水平的样品。采用紫外光谱、荧光光谱、凝胶渗透色谱（GPC）、动态光散射（DLS）、十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）、免疫印迹（Western blotting）和酶联免疫吸附测定（ELISA）等方法对其进行表征。在野生型小鼠和转基因小鼠中对hIFN α 2免疫耐受进行免疫原性评价。ELISA检测血清中是否存在rhIFN α 2b特异性抗体。氧化和老化的制备在团聚体的水平和性质上有很大的不同。与天然rhIFNa2b相比，所有含有聚集体的制剂都增加了野生型小鼠的免疫应答，并能够打破转基因小鼠的耐受性。聚集的蛋白质的构象越像天然的，在转基因小鼠中的免疫原性越强。通过金属催化制备的天然聚集体在转基因小鼠中诱导了剂量依赖性的耐受性丧失。总之，转基因小鼠模型可用于筛选在加速储存条件下获得的低水平免疫原性聚集体的rhIFNa2b配方。(3)中国医药科学，2006,(5):391 - 391
The aim of this study was to determine the sensitivity of transgenic immune tolerant mice for the type and level of aggregation of recombinant human interferon alpha2b (rhIFN alpha 2b). RhIFN alpha 2b was aggregated by metal-catalyzed oxidation or by incubation at elevated temperature and various pHs. Native rhIFNa2b was mixed with oxidized rhIFNa2b at different ratios to obtain samples with different aggregation levels. The preparations were characterized by UV and fluorescence spectroscopy, gel permeation chromatography (GPC), dynamic light scattering (DLS), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting, and ELISA. The immunogenicity was evaluated in wild-type mice and transgenic mice immune tolerant for hIFN alpha 2. Sera were analyzed by ELISA for the presence of rhIFN alpha 2b-specific antibodies. The oxidized and aged preparations widely differed regarding the level and nature of aggregates. All preparations containing aggregates increased the immune response in the wild-type mice as compared to native rhIFNa2b and were able to break the tolerance of the transgenic mice. The more native-like the conformation of the aggregated proteins, the more immunogenic the preparations were in the transgenic mice. The native-like aggregates prepared via metal catalysis induced a dose-dependent loss of tolerance in the transgenic mice. In conclusion, the transgenic mouse model can be used to screen rhIFNa2b formulations for low levels of immunogenic aggregates obtained under accelerated storage conditions. (c) 2006 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 95:1084-1096, 2006