Fluorescence-based quantitative methods for detecting human immunodeficiency virus type 1-induced syncytia.
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.
Department of Internal Medicine, The University of Iowa, Iowa City, Iowa 52242, USA.
S
S
Wünschmann
J T
JT
Stapleton
D004279:DNA, Viral / Q000378:metabolism; D005434:Flow Cytometry; D015726:Giant Cells / Q000502:physiology*; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D008856:Microscopy, Fluorescence; D011419:Propidium; D013194:Staining and Labeling; D014407:Tumor Cells, Cultured
D004279:DNA, Viral; D011419:Propidium