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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Detection of Antiendothelial Cell Antibodies by an Enzyme-Linked Immunosorbent Assay Using Antigens from Cell Lysate: Minimal Interference with Antinuclear Antibodies and Rheumatoid Factors
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Detection of Antiendothelial Cell Antibodies by an Enzyme-Linked Immunosorbent Assay Using Antigens from Cell Lysate: Minimal Interference with Antinuclear Antibodies and Rheumatoid Factors

机译：使用细胞裂解液中的抗原通过酶联免疫吸附法检测抗内皮细胞抗体：对抗核抗体和类风湿因子的干扰最小

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摘要

The objective of the present work was to set up a routine test adapted to screening for antiendothelial cell antibodies (AECAs) in serum samples with minimal interference from antinuclear antibodies (ANAs) or rheumatoid factors (RFs). We compared the titers of AECAs titrated following two enzyme-linked immunosorbent assays (ELISAs): (i) an ELISA with ethanol-fixed EA.hy926 monolayers as the antigenic substrate and (ii) an ELISA with nucleus-depleted lysates prepared from EA.hy926 cells and normalized for protein (1.0 to 1.7 mg/ml) and DNA (≤0.1 μg/ml) contents as a surrogate substrate (postnuclear supernatant ELISA [PNS-ELISA]). The AECA titers in 51 serum samples, including 28 samples containing ANAs, were compared. A significantly positive correlation (r = 0.77; P < 0.001) between the two series was shown only for the ANA-negative serum samples. Conversely, ANAs or RFs in samples were shown not to interfere in tests for AECAs by the PNS-ELISA. AECAs recognize their antigenic targets in postnuclear supernatants, which is representative of the endothelial antigenic content, with improvement of the reliability of the assay, a prerequisite to application of the assay for their evaluation in clinical practice.
机译：本工作的目的是建立一种常规测试，以筛查血清样品中的抗内皮细胞抗体（AECA），而不受抗核抗体（ANA）或类风湿因子（RF）的干扰最小。我们比较了通过两种酶联免疫吸附测定（ELISA）滴定的AECA的效价：（i）以乙醇固定的EA.hy926单层作为抗原底物的ELISA和（ii）用从EA制备的去核裂解物的ELISA。 hy926细胞，将蛋白质（1.0至1.7 mg / ml）和DNA（≤0.1μg/ ml）含量标准化为替代底物（核后上清ELISA [PNS-ELISA]）。比较了51个血清样品（包括28个含ANAs样品）的AECA滴度。仅对于ANA阴性血清样品，两个系列之间显示出显着正相关（ r = 0.77; P <0.001）。相反地​​，样品中的ANAs或RFs不会干扰PNS-ELISA对AECA的测试。 AECA在核后上清液中识别其抗原靶标，这是内皮抗原含量的代表，并提高了测定的可靠性，这是在临床实践中应用测定进行评估的先决条件。

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