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Fluorescence-based quantitative methods for detecting human immunodeficiency virus type 1-induced syncytia.

S Wünschmann1, J T Stapleton

  • 1Department of Internal Medicine, The University of Iowa, Iowa City, Iowa 52242, USA.

Journal of Clinical Microbiology
|August 2, 2000
PubMed
Summary

New fluorescence assays accurately detect and quantify human immunodeficiency virus type 1 (HIV-1) induced cell fusion (syncytia). These methods enable precise measurement of HIV replication and anti-HIV drug efficacy.

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Area of Science:

  • Virology
  • Cell Biology
  • Immunology

Background:

  • Cell fusion, or syncytia formation, is a hallmark of human immunodeficiency virus type 1 (HIV-1) infection.
  • Current methods for assessing syncytia lack quantification and do not estimate the number of cells involved in fusion.

Purpose of the Study:

  • To develop and validate novel fluorescence-based methods for detecting and quantifying HIV-1-induced syncytia in vitro.
  • To assess the utility of these assays for evaluating antiretroviral compounds.

Main Methods:

  • Two fluorescence-based methods were employed: DNA staining with propidium iodide for flow cytometry analysis of cell size, and two-color cytoplasmic staining for fluorescence microscopy.
  • Lymphoblastoid cell lines (MT-2, SupT1) were infected with syncytium-inducing (SI) HIV-1 isolates.

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Main Results:

  • Both fluorescence assays successfully detected and quantified HIV-induced syncytia.
  • The methods differentiated between SI and non-SI HIV isolates and were applicable to multiple CD4(+) T-cell lines.
  • Small syncytia were identifiable using the two-color cytoplasmic staining method.
  • The assays accurately assessed the inhibitory effects of antiretroviral drugs like zidovudine and zalcitabine.

Conclusions:

  • Fluorescence-based assays provide a rapid, practical, and quantitative approach to measure HIV-1 syncytia formation.
  • These assays are valuable tools for studying HIV replication and screening potential anti-HIV compounds.