韦格纳肉芽肿病自身抗原蛋白酶3的抗原决定簇研究

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 2 2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 %。 2 2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 (2 7% )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。

Epitope mapping on Wegener's granulomatosis autoantigen proteinase 3

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