目的：在臨床中，存在部分患者有提示APS的臨床症狀而傳統抗磷脂抗體檢測持續陰性。對於這部分病例，我們稱之為血清陰性抗磷脂綜合征(SN-APS)。本文探索薄層色譜分析(thin-layerchromatography，TLC)免疫染色法在檢測SN-APS患者血清中抗體的應用意義。
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陽性患者的血清，以及32名健康對照的血清，應用TLC免疫染色法檢測抗磷脂抗體。應用ELISA方法檢測抗β2GP1抗體，抗膜聯蛋白A2，抗膜聯蛋白A5抗體，和抗凝血酶原抗體。Eahy926, 一個來自人體的內皮細胞係，用SN-APS患者的血清IgG成分孵育後，應用Western blot分析磷脂-白介素-1受體相關激酶(IRAK)和磷脂-NFκB，應用細胞熒光分析檢測分VCAM-1，應用ELISA檢測上清中組織因子(TF)。
結果：TLC免疫染色法在58.3%SN-APS患者中檢測到aPL：抗心磷脂抗體47.2%，抗溶血磷脂抗體41.7%，抗磷脂酰乙醇胺抗體30.5%。36名患者中有6人抗膜聯蛋白抗體陽性。 Eahy926細胞係經SN-APS患者血清IgG孵育後誘導IRAK磷酸化，NF-κB激活，VCAM-1表麵表達及TF釋放。
結論：TLC免疫染色法能夠識別SN-APS患者血清中的aPL。此外，研究結果提示這些抗體在體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.
