Chen Jie, Chen Min, Liu Zhang suo, Zhao Ming hui
Renal Division, Peking University First Hospital, Beijing 100034, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2008 Apr;40(2):155-9.
To investigate the inhibitory effects on myeloperoxidase (MPO) oxidation activity by affinity-purified anti-MPO antibodies from patients with microscopic polyangiitis (MPA) and to further investigate the interaction between MPO, ceruloplasmin and anti-MPO antibodies.
Human IgG fractions were purified from plasma of 11 patients with anti-MPO antibody positive MPA and sera of 12 normal controls. Anti-MPO antibodies were further purified from anti-MPO antibody containing IgG fractions using MPO affinity chromatography. The enzyme activity of MPO was measured, in the presence of anti-MPO antibodies and normal IgG preparations, using a classical MPO oxidation assay. Interaction between ceruloplasmin, MPO and anti-MPO antibodies was further investigated using ELISA.
Anti-MPO antibodies from 7/11 patients with MPA could inhibit the MPO activity as non-competitive inhibitors in a dose-dependent manner. Ceruloplasmin could competitively inhibit the oxidation activity of MPO in a dose-dependent and time-dependent manner. Anti-MPO antibodies could inhibit the binding between MPO and ceruloplasmin to a maximum of (75.4+/-11.6)%.
Anti-MPO antibodies, from the majority of patients with MPA, could inhibit the oxidation activity of MPO and interfere with the binding between MPO and ceruloplasmin.
研究显微镜下多血管炎(MPA)患者亲和纯化的抗髓过氧化物酶(MPO)抗体对MPO氧化活性的抑制作用,并进一步研究MPO、铜蓝蛋白与抗MPO抗体之间的相互作用。
从11例抗MPO抗体阳性的MPA患者血浆和12例正常对照者血清中纯化人IgG组分。使用MPO亲和层析从含抗MPO抗体的IgG组分中进一步纯化抗MPO抗体。在存在抗MPO抗体和正常IgG制剂的情况下,采用经典的MPO氧化试验测定MPO的酶活性。使用酶联免疫吸附测定法(ELISA)进一步研究铜蓝蛋白、MPO和抗MPO抗体之间的相互作用。
7/11例MPA患者的抗MPO抗体可作为非竞争性抑制剂以剂量依赖性方式抑制MPO活性。铜蓝蛋白可呈剂量依赖性和时间依赖性地竞争性抑制MPO的氧化活性。抗MPO抗体可将MPO与铜蓝蛋白之间的结合抑制至最大(75.4±11.6)%。
大多数MPA患者的抗MPO抗体可抑制MPO的氧化活性并干扰MPO与铜蓝蛋白之间的结合。