系统性血管炎抗内皮细胞抗体与抗中性粒细胞胞质抗体的相关性初探

Analysis of the correlation between anti-endothelial cell antibodies and antineutrophil cytoplasmic antibodies in systemic vasculitides

目的以来源于大小血管内皮细胞的两种永生细胞株为底物,检测系统性血管炎血清中抗内皮细胞抗体(AECA),分析其与抗中性粒细胞胞质抗体(ANCA)的相关性。方法细胞酶联免疫吸附试验(Cyto-ELISA)法检测血清中AECA;间接免疫荧光法(IIF)及抗抗体结合内皮细胞表面的蛋白酶3(PR3)、抗MPO-ELISA检测血清中ANCA;IIF及抗PR3、抗MPO-ELISA检测细胞株中PR3及MPO;反转录—聚合酶链反应(RT-PCR)法检测细胞株中PR3及MPOmRNA。结果AECAEA(EA.hy926为底物所测AECA)阳性率33.6%(41/122),AECAHMEC(HMEC-1为底物所测AECA)37.7%(46/122),ANCA35.3%(43/122),AECAEA或AECAHMEC与ANCA串联诊断系统性血管炎敏感性分别为59.8%(73/122)或60.7%(74/122)。AECAEA与ANCA,AECAHMEC与ANCA分别行配对字2检验,差异均无显著性(P>0.05),符合率仅分别为49.2%及51.6%。EA.hy926和HMEC-1中蛋白水平及mRNA水平均无PR3和MPO的表达。结论EA.hy926与HMEC-1中无PR3、MPO蛋白水平的表达,ANCA与AECA可能是两种相互独立的抗体,串联检测可提高诊断敏感性。

Objective To study the presence of anti-endothelial cell antibodies (AECA) in systemic vasculitides by cyto-ELISA in two immortalized cell line substrates, EA.hy926 and HMEC-1, which were derived from macro- and micro-vessel endothelial cellsthe correlation between AECA and antineutrophil cytoplasmic antibodies (ANCA) was assessed. Methods AECA was detected by cyto-ELISA, ANCA by IIF, anti-PR3 and anti-MPO by ELISA. PR3 and MPO in EA.hy926 and HMEC-1 were tested by IIF and anti-PR3 while anti-MPO was examined by ELISA. PR3 and MPO mRNA in EA.hy926 and HMEC-1 were detected by RT-PCR. Results The positivity of AECAEA, AECAHMEC and ANCA was 33.6%(41/122), 37.7%(46/122), 35.3%(43/122) respectively. No significant correlaion between AECA and ANCA was observed (P>0.05). The matching rate between AECAEA and ANCA was only 49.2%, and 51.6% betwere AECAHMEC and ANCA.EA.hy926 and HMEC-1 did not express neither PR3, MPO protein nor their mRNA. Conclusion EA.hy926 and HMEC-1 do not express PR3 or MPO protein or their mRNA. ANCA and AECA are two independent antibodies. Combining the detection of these two antibodies can raise the diagnostic sensitivity of systemic vasculitides.