Wagatsuma M, Asami N, Miyachi J, Uchida S, Watanabe H, Amann E
Hoechst Japan Limited, Pharma Research Laboratories, Kawagoe, Saitama, Japan.
Mol Immunol. 1993 Nov;30(16):1491-8. doi: 10.1016/0161-5890(93)90457-m.
Five human nuclear antigens, RNP 70 kD, SS-A, SS-B, Sm-B and Sm-D, were produced in E. coli using the expression vector pSEM. cDNAs encoding these antigens were ligated to a truncated lacZ' gene of the vector and the beta-galactosidase fusion proteins were efficiently expressed as intracellular inclusion bodies after isopropyl-beta-thiogalactopyranoside induction. The antibody reactivities of these fusion proteins were evaluated by Western blot and by ELISA employing panel sera from patients with autoimmune diseases such as systemic lupus erythematosus, Sjögren's syndrome or mixed connective tissue disease. The three fusion proteins, RNP 70 kD, SS-B, and Sm-B, showed good reactivities in both systems, whereas the other two fusion proteins, SS-A and Sm-D, showed poor and no reactivity in both systems, respectively. It can be concluded that RNP 70 kD, SS-B and Sm-B recombinant antigens are useful reagents for the differential diagnosis of the autoimmune diseases.
利用表达载体pSEM在大肠杆菌中产生了5种人类核抗原，即核糖核蛋白70kD、干燥综合征-A抗原（SS-A）、干燥综合征-B抗原（SS-B）、Sm-B和Sm-D。编码这些抗原的cDNA被连接到载体的截短lacZ'基因上，异丙基-β-D-硫代半乳糖苷诱导后，β-半乳糖苷酶融合蛋白作为细胞内包涵体高效表达。采用来自系统性红斑狼疮、干燥综合征或混合性结缔组织病等自身免疫性疾病患者的一组血清，通过蛋白质免疫印迹法和酶联免疫吸附测定法评估这些融合蛋白的抗体反应性。三种融合蛋白，即核糖核蛋白70kD、SS-B和Sm-B，在这两种检测系统中均表现出良好的反应性，而另外两种融合蛋白，即SS-A和Sm-D，在这两种检测系统中分别表现出较差的反应性和无反应性。可以得出结论，核糖核蛋白70kD、SS-B和Sm-B重组抗原是自身免疫性疾病鉴别诊断的有用试剂。
