目的：在临床中，存在部分患者有提示APS的临床症状而传统抗磷脂抗体检测持续阴性。对于这部分病例，我们称之为血清阴性抗磷脂综合征(SN-APS)。本文探索薄层色谱分析(thin-layerchromatography，TLC)免疫染色法在检测SN-APS患者血清中抗体的应用意义。

        方法：收集36名SN-APS患者，19名APS患者，及18名SLE患者，20名抗HCV阳性患者的血清，以及32名健康对照的血清，应用TLC免疫染色法检测抗磷脂抗体。应用ELISA方法检测抗β2GP1抗体，抗膜联蛋白A2，抗膜联蛋白A5抗体，和抗凝血酶原抗体。Eahy926, 一个来自人体的内皮细胞系，用SN-APS患者的血清IgG成分孵育后，应用Western blot分析磷脂-白介素-1受体相关激酶(IRAK)和磷脂-NFκB，应用细胞荧光分析检测分VCAM-1，应用ELISA检测上清中组织因子(TF)。

        结果：TLC免疫染色法在58.3%SN-APS患者中检测到aPL：抗心磷脂抗体47.2%，抗溶血磷脂抗体41.7%，抗磷脂酰乙醇胺抗体30.5%。36名患者中有6人抗膜联蛋白抗体阳性。 Eahy926细胞系经SN-APS患者血清IgG孵育后诱导IRAK磷酸化，NF-κB激活，VCAM-1表面表达及TF释放。

        结论：TLC免疫染色法能够识别SN-APS患者血清中的aPL。此外，研究结果提示这些抗体在体外有促炎及促凝的作用。

        附全文：

        In clinical practice it is possible to find patients withclinical signs suggestive ofanti-phospholipidsyndrome(APS)who are persistently negative for the routinelyusedanti-phospholipidantibodies(aPL). Therefore, the termproposed for these cases wasseronegativeAPS (SN-APS). Weinvestigated the clinical usefulnessofthin-layerchromatography(TLC)immunostainingindetectingserumaPL in patients presenting clinical features of SN-APS. Sera from 36 patientswith SN-APS, 19 patients with APS, 18 patients with systemic lupuserythematosus (SLE), 20 anti-hepatitis C virus (HCV)-positive subjects and 32healthy controls were examined for aPL using TLCimmunostaining. Anti-β(2) -glycoprotein-I,anti-annexin II, anti-annexin V and anti-prothrombinantibodiesweretested by enzyme-linked immunosorbent assays (ELISA). Eahy926, a human-derivedendothelial cell line, was incubated with immunoglobulin (Ig)G fraction fromSN-APS patients and analysis of phospho-interleukin (IL)-1 receptor-associatedkinase (IRAK) and phospho-nuclear factor (NF)-κB wasperformed by Western blot, vascular cell adhesion molecule 1 (VCAM-1)expression by cytofluorimetric analysis and supernatants tissue factor (TF)levels by ELISA. TLCimmunostainingshowed aPL in 58·3% of SN-APS patients: anti-cardiolipin in 47·2%, anti-lyso(bis)phosphatidic acid in 41·7%and anti-phosphatidylethanolamine in 30·5%. Six of 36patients showed anti-annexin II. Incubation of Eahy926 cells with IgG fromSN-APS induced IRAK phosphorylation, NF-κB activation,VCAM-1 surface expression and TF cell release.TLCimmunostainingcould identify the presence of aPL in patientswith SN-APS. Moreover, the results suggest the proinflammatory and procoagulanteffects in vitro of theseantibodies.

        引自：Conti Fet al.,Thin-layerchromatographyimmunostainingindetectinganti-phospholipidantibodiesinseronegativeanti-phospholipidsyndrome.Clin Exp Immunol.2012Mar;167(3):429-37. doi: 10.1111/j.1365-2249.2011.04532.x.