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首页> 外文期刊>Journal of Clinical Microbiology >Fluorescence-Based Quantitative Methods for Detecting Human Immunodeficiency Virus Type 1-Induced Syncytia
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Fluorescence-Based Quantitative Methods for Detecting Human Immunodeficiency Virus Type 1-Induced Syncytia

机译：基于荧光的定量方法检测人类免疫缺陷病毒1型引起的合胞体

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Cell fusion induced by human immunodeficiency virus type 1 (HIV-1) is usually assessed by counting multinucleated giant cells (syncytia) visualized by light microscopy. Currently used methods do not allow quantification of syncytia, nor do they estimate the number of cells involved in cell fusion. We describe two fluorescence-based methods for the detection and quantification of HIV-1-induced in vitro syncytium formation. The lymphoblastoid cell lines MT-2 and SupT1 were infected with syncytium-inducing (SI) HIV-1 isolates. Syncytia were detected by DNA staining with propidium iodide using flow cytometry to determine cell size or by two-color cytoplasmic staining of infected cell populations by using fluorescence microscopy. Both methods were able to detect and quantify HIV-induced syncytia. The methods could distinguish between SI and non-SI HIV isolates and could be used with at least two separate types of CD4+ T-cell lines. Small syncytia can be readily identified by the two-color cytoplasmic staining method. Both methods were also shown to be useful for evaluating antiretroviral compounds, as demonstrated by the accurate assessment of HIV inhibition by azidothymidine (zidovudine), dideoxycytidine (zalcytibine), and hydroxyurea. These fluorescence-based assays allow a rapid and practical method for measuring HIV replication and anti-HIV activity of potential inhibitory compounds.
机译：通常通过对光学显微镜下可见的多核巨细胞（合胞体）计数来评估由1型人类免疫缺陷病毒（HIV-1）诱导的细胞融合。当前使用的方法不能定量合胞体，也不能估计参与细胞融合的细胞数量。我们描述了两种基于荧光的方法来检测和定量HIV-1诱导的体外合胞体形成。淋巴母细胞细胞系MT-2和SupT1被合胞体诱导（SI）HIV-1分离株感染。合胞体通过流式细胞术用碘化丙啶进行DNA染色来确定细胞大小，或者通过使用荧光显微镜对感染的细胞群进行双色细胞质染色来检测。两种方法都能够检测和量化HIV引起的合胞体。该方法可以区分SI和非SI HIV分离株，并且可以与至少两种不同类型的CD4 + T细胞系一起使用。通过两种颜色的细胞质染色方法可以很容易地识别出小的合胞体。这两种方法还显示出可用于评估抗逆转录病毒化合物，如通过叠氮胸苷（齐多夫定），双脱氧胞苷（扎西替丁）和羟基脲对HIV抑制作用的准确评估所证明的那样。这些基于荧光的测定法提供了一种快速而实用的方法来测量潜在抑制性化合物的HIV复制和抗HIV活性。

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