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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。
Objective To study the antigenic determinants of anti-protease 3 autoantibodies in the serum of patients with Wegener’s granulomatosis. Methods Western blotting was used to study the antigenic activity of proteinase 3 under reducing and non-reducing conditions. Using competitive inhibition ELISA and immunoblot analysis, the two anti-protease 3 monoclonal antibodies between HZ1F12 and HZ1H3, and between the monoclonal antibody and 22 active stages of Wegener’s granuloma anti-protease 3 serum, Competitive inhibition of protease 3, and analysis of its possible antigenic determinants. Results Both serum and monoclonal antibodies recognized protease 3 under non-reducing conditions. HZ1F12 can be inhibited by HZ1H3 by 74%. Of the 2 2 serums, 10 (46%) were completely or partially inhibited by HZ1F12, 9 (41%) were partially inhibited by HZ1H3 and 6 (27%) were inhibited by both HZ1F12 and HZ1H3. Monoclonal antibodies can also significantly inhibit the patient’s serum binding capacity. Conclusion The determinants of the autoantibodies of Wegener’s granulomatosis are three-dimensional configuration. Monoclonal antibodies HZ1F12 and HZ1H3 recognize similar or similar antigenic determinants. Antigen determinants recognized by anti-protease 3-positive sera from patients with Wegener’s granulomatosis were inconsistent.