Pluschke G, Jenni L, van Alphen L, Lefkovits I
Clin Exp Immunol. 1986 Nov;66(2):331-9.
The early stages of inflammatory responses are characterized by the rapid synthesis of a heterogenous group of plasma proteins known as acute phase reactants. We show that the complex changes in the serum protein composition of mice in response to infections can easily be analysed by two-dimensional (2D) gel electrophoresis, which allows simultaneous analysis of numerous proteins from small volume samples. While changes in the protein composition can be visualized by silver staining, radiofluorography of gels loaded with serum samples from mice that had been labelled in vivo with 35S-methionine allows an analysis of the changes in the pattern of protein synthesis. Thus, these techniques should allow the evaluation of the relative contributions of alterations of protein synthesis and catabolism to the changes in the overall concentration of individual acute phase reactants. Identification of proteins in the 2 D gel pattern can be easily accomplished by co-electrophoresing small serum samples together with immunoprecipitates obtained from in-vivo labelled serum. Using this approach we were able to identify some of the major acute phase reactants of mice. Some of these proteins, like haptoglobin and haemopexin, show concentration increases that are characteristic for type III reactants like C-reactive protein (CRP) or serum amyloid A component (SAA) in man. Results obtained with serum from healthy and infected human newborns indicate that 2D gel electrophoresis could be used to analyse changes in human plasma protein profiles, which would make it a valuable tool for diagnosis and management in certain clinical situations.
炎症反应的早期阶段以快速合成一组异质性血浆蛋白为特征，这些蛋白被称为急性期反应物。我们发现，通过二维（2D）凝胶电泳可以轻松分析小鼠血清蛋白组成因感染而发生的复杂变化，该技术能够同时分析少量样品中的多种蛋白质。虽然蛋白质组成的变化可以通过银染可视化，但对用35S-甲硫氨酸体内标记的小鼠血清样品进行凝胶放射自显影，可以分析蛋白质合成模式的变化。因此，这些技术应该能够评估蛋白质合成和分解代谢的改变对单个急性期反应物总体浓度变化的相对贡献。通过将少量血清样品与从体内标记血清中获得的免疫沉淀物共同电泳，可以轻松完成二维凝胶图谱中蛋白质的鉴定。使用这种方法，我们能够鉴定出小鼠的一些主要急性期反应物。其中一些蛋白质，如触珠蛋白和血红素结合蛋白，其浓度升高具有人类中III型反应物（如C反应蛋白（CRP）或血清淀粉样蛋白A成分（SAA））的特征。从健康和受感染的人类新生儿血清中获得的结果表明，二维凝胶电泳可用于分析人类血浆蛋白谱的变化，这将使其成为某些临床情况下诊断和管理的有价值工具。
