Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy

Benjamin Dale, Gregory P. McNerney, Deanna L. Thompson, Wolfgang Hübner, Thomas R Huser, Benjamin K. Chen

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

By fusing the green fluorescent protein to their favorite proteins, biologists now have the ability to study living complex cellular processes using fluorescence video microscopy. To track the movements of the human immunodeficiency virus core protein during cell-to-cell transmission of human immunodeficiency virus, we have GFP-tagged the Gag protein in the context of an infectious molecular clone of HIV, called HIV Gag-iGFP. We study this viral clone using video confocal microscopy. In the following visualized experiment, we transfect a human T cell line with HIV Gag-iGFP, and we use fluorescently labeled uninfected CD4+ T cells to serve as target cells for the virus. Using the different fluorescent labels we can readily follow viral production and transport across intercellular structures called virological synapses. Simple gas permeable imaging chambers allow us to observe synapses with live confocal microscopy from minutes to days. These approaches can be used to track viral proteins as they move in from one cell to the next.

Original languageEnglish (US)
Article numbere2061
JournalJournal of Visualized Experiments
Issue number44
DOIs
StatePublished - Oct 2010

Fingerprint

T-cells
Confocal microscopy
Viruses
Confocal Microscopy
Clone Cells
HIV
Viral Core Proteins
Proteins
gag Gene Products
Video Microscopy
Viral Proteins
Green Fluorescent Proteins
Synapses
Labels
Microscopic examination
Gases
Fluorescence
Human Immunodeficiency Virus Proteins
T-Lymphocytes
Imaging techniques

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

Dale, B., McNerney, G. P., Thompson, D. L., Hübner, W., Huser, T. R., & Chen, B. K. (2010). Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy. Journal of Visualized Experiments, (44), [e2061]. https://doi.org/10.3791/2061

Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy. / Dale, Benjamin; McNerney, Gregory P.; Thompson, Deanna L.; Hübner, Wolfgang; Huser, Thomas R; Chen, Benjamin K.

In: Journal of Visualized Experiments, No. 44, e2061, 10.2010.

Research output: Contribution to journalArticle

Dale, Benjamin ; McNerney, Gregory P. ; Thompson, Deanna L. ; Hübner, Wolfgang ; Huser, Thomas R ; Chen, Benjamin K. / Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy. In: Journal of Visualized Experiments. 2010 ; No. 44.
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