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首页> 外文期刊>Journal of Virological Methods >Measles virus fusion protein- and hemagglutinin-transfected cell lines are a sensitive tool for the detection of specific antibodies by a FACS-measured immunofluorescence assay.
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Measles virus fusion protein- and hemagglutinin-transfected cell lines are a sensitive tool for the detection of specific antibodies by a FACS-measured immunofluorescence assay.

机译：麻疹病毒融合蛋白和血凝素转染的细胞系是通过FACS测量的免疫荧光检测来检测特定抗体的灵敏工具。

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摘要

A FACS-measured immunofluorescence assay was developed for the detection of antibodies directed against the hemagglutinin (H) and fusion (F) glycoproteins of measles virus (MV). Human melanoma cell lines transfected with either the MV H or F genes, which showed a high surface expression of the respective proteins in their native conformation, were used as target cells. The cells were incubated with diluted plasma samples, and stained subsequently with FITC-conjugated secondary antibodies. The FACS-measured fluorescence signals correlated directly with the amount of specific immunoglobulins over a wide concentration range. The use of different conjugates enabled the separate detection of MV-specific IgG, IgM, IgA and IgG subclasses, with relatively low backgrounds. Hemagglutinin-specific IgG, IgM and IgA fluorescence signals were shown to correlate well with MV-specific IgG ELISA titers and MV-specific IgM or IgA capture ELISA OD450-values, respectively. The polyclonal conjugates with specificity for human immunoglobulins offered sufficient cross-reactivity to detect MV-specific IgG, IgM and IgA in plasma samples of cynomolgus macaques, making this technique a useful tool for studying serological responses in vaccination and challenge experiments in non-human primate models.
机译：开发了一种FACS测量的免疫荧光测定法，用于检测针对麻疹病毒（MV）的血凝素（H）和融合（F）糖蛋白的抗体。将以MV H或F基因转染的人黑素瘤细胞系（以其天然构象显示各自蛋白的高表面表达）用作靶细胞。将细胞与稀释的血浆样品一起孵育，并随后用缀有FITC的二抗染色。 FACS测量的荧光信号与很宽的浓度范围内特异性免疫球蛋白的量直接相关。使用不同的结合物可以分别检测背景相对较低的MV特异性IgG，IgM，IgA和IgG亚类。已显示血凝素特异性IgG，IgM和IgA荧光信号分别与MV特异性IgG ELISA滴度和MV特异性IgM或IgA捕获ELISA OD450值密切相关。对人免疫球蛋白具有特异性的多克隆偶联物提供了足够的交叉反应性，可检测食蟹猕猴血浆样品中的MV特异性IgG，IgM和IgA，从而使该技术成为研究疫苗接种和非人灵长类动物攻击实验中血清学反应的有用工具。楷模。

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