摘要目的研究韦格纳肉芽肿病患者血清中抗蛋白酶3自身抗体的抗原决定簇.方法应用免疫印记分析法,研究在还原和非还原条件下蛋白酶3的抗原活性.应用竞争性抑制ELISA和免疫印记分析法,研究两个抗蛋白酶3的单克隆抗体HZ1F12和HZ1H3之间,及单克隆抗体和22份韦格纳肉芽肿活动期的抗蛋白酶3血清之间,对蛋白酶3的竞争抑制能力,并分析其可能的抗原决定簇.结果患者血清及单克隆抗体均识别非还原条件下的蛋白酶3.HZ1F12可以被HZ1H3抑制74%.22份血清中,10份(46%)可以完全或部分被HZ1F12抑制,9份(41%)可以部分被HZ1H3抑制,6份(27%)可以被HZ1F12和HZ1H3二者抑制.单克隆抗体也可以明显抑制患者血清的结合能力.结论韦格纳肉芽肿病自身抗体的抗原决定簇是三维立体构型.单克隆抗体HZ1F12和HZ1H3识别相似或相近的抗原决定簇.韦格纳肉芽肿病患者的抗蛋白酶3阳性血清识别的抗原决定簇是不一致的.

abstractsAbstract:Objective To map the epitopes on Wegener's granulomatosis autoantigen proteinase 3.Methods Antigenicity of proteinase 3 was studied with Western blot analysis in which proteinase 3 was prepared under reducing and non-reducing conditions. Two anti-proteinase 3 monoclonal antibodies, HZ1F12 and HZ1H3, were used to inhibit each other and to inhibit 22 anti-proteinase 3 positive sera from patients with Wegener's granulomatosis in competitive inhibition enzyme-linked immunosorbent assays (ELISA) and Western blot analysis. Results All monoclonal antibodies and patient sera recognized proteinase 3 under non-reducing conditions in Western blot analysis. HZ1F12 was inhibited 74% by HZ1H3. 10/22 (46%) sera were completely or partially inhibited by HZ1F12; 9/22 (41%) sera were partially inhibited by HZ1H3; and 6/22 (27%) were inhibited by both monoclonal antibodies. In inhibition Western blot analysis, the binding of patient sera to proteinase 3 could be inhibited by HZ1F12. Conclusions The epitopes of Wegener's granulomatosis autoantigen were conformational. Anti-proteinase 3 monoclonal HZ1F12 and HZ1H3 recognized similar or overlapping epitopes on the proteinase 3 molecule. Epitopes of proteinase 3 recognized by anti-proteinase 3 positive sera were not restricted.