Van den Bergh Karolien, Hooijkaas Herbert, Blockmans Daniel, Westhovens René, Op De Beéck Katrijn, Verschueren Patrick, Dufour Diana, van de Merwe Joop P, Fijak Monika, Klug Jörg, Michiels Georges, Devogelaere Benoit, De Smedt Humbert, Derua Rita, Waelkens Etienne, Blanckaert Norbert, Bossuyt Xavier
Laboratory Medicine, Immunology, University Hospitals Leuven, Leuven, Belgium.
Clin Chem. 2009 May;55(5):946-54. doi: 10.1373/clinchem.2008.115626. Epub 2009 Mar 5.
Serum samples from patients with autoimmune connective tissue diseases that show a finely speckled antinuclear antibody (ANA) on indirect immune-fluorescence often have antibodies against unknown nuclear target antigens. To search for such autoantigens we applied a proteomic approach using sera from patients with a high ANA titer (>or=640) and finely speckled fluorescence but in whom no antibodies to extractable nuclear antigens (ENA) could be identified.
Using an immunoproteomics approach we identified heterogeneous nuclear ribonucleoprotein H1 (hnRNP H1) as a novel nuclear target of autoantibody response.
Recombinant rat hnRNP H1 reacted in Western blot analyses with 48% of 93 sera from patients with primary Sjögren syndrome and with 5.2% of 153 sera from patients with other connective tissue diseases (diseased controls). For comparison, the diagnostic sensitivity and specificity of anti-Sjögren syndrome A (SSA) antibodies for primary Sjögren syndrome in the same patient cohort were 88.2% and 76.3%, respectively. Interestingly, 5 of 11 primary Sjögren syndrome patients with no anti-SSA or anti-SSB antibodies had anti-hnRNP H1 antibodies. Anti-hnRNP H1 antibodies were preabsorbed by hnRNP H1, as demonstrated by indirect immunofluorescence. In an evaluation of the presence of anti-hnRNP H1 antibodies in 188 consecutive samples submitted to the clinical laboratory with positive ANA (titer >or=160), anti-hnRNP H1 antibodies were found in 3 of 7 (2 primary and 5 secondary) Sjögren syndrome patients and in 8.3% of the diseased controls.
HnRNP H1 is a newly discovered autoantigen that could become an additional diagnostic marker.
自身免疫性结缔组织病患者的血清样本在间接免疫荧光检测中显示出细颗粒状抗核抗体(ANA),这些样本中常常含有针对未知核靶抗原的抗体。为了寻找此类自身抗原,我们采用蛋白质组学方法,使用来自ANA滴度高(≥640)且荧光呈细颗粒状,但未检测到可提取核抗原(ENA)抗体的患者血清。
我们采用免疫蛋白质组学方法,将不均一核核糖核蛋白H1(hnRNP H1)鉴定为自身抗体反应的一种新的核靶标。
重组大鼠hnRNP H1在蛋白质印迹分析中与93例原发性干燥综合征患者血清中的48%发生反应,与153例其他结缔组织病患者(疾病对照)血清中的5.2%发生反应。作为比较,同一患者队列中抗干燥综合征A(SSA)抗体对原发性干燥综合征的诊断敏感性和特异性分别为88.2%和76.3%。有趣的是,11例无抗SSA或抗SSB抗体的原发性干燥综合征患者中有5例具有抗hnRNP H1抗体。间接免疫荧光显示,抗hnRNP H1抗体可被hnRNP H1预先吸附。在对188份ANA阳性(滴度≥160)并提交至临床实验室的连续样本进行抗hnRNP H1抗体存在情况评估时,在7例(2例原发性和5例继发性)干燥综合征患者中的3例以及8.3%的疾病对照中发现了抗hnRNP H1抗体。
HnRNP H1是一种新发现的自身抗原,可能成为一种额外的诊断标志物。