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

Dale B, McNerney GP, Thompson DL, Hübner W, Huser T, Chen BK (2010)
Journal of visualized experiments : JoVE (44): 2061.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
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.
Stichworte
Jurkat Cells; Microscopy; Humans; HIV Infections/virology; HIV Infections/transmission; HIV/physiology; HIV/metabolism; HIV/genetics; Green Fluorescent Proteins/metabolism; Green Fluorescent Proteins/analysis; CD4-Positive T-Lymphocytes/physiology; Confocal/methods; CD4-Positive T-Lymphocytes/virology; Human Immunodeficiency Virus/metabolism; gag Gene Products; Human Immunodeficiency Virus/analysis; gag Gene Products; Transfection; Microscopy; Video/methods
Erscheinungsjahr
2010
Zeitschriftentitel
Journal of visualized experiments : JoVE
Ausgabe
44
Seite(n)
2061
ISSN
1940-087X
eISSN
1940-087X
Page URI
https://pub.uni-bielefeld.de/record/2576779

Zitieren

Dale B, McNerney GP, Thompson DL, Hübner W, Huser T, Chen BK. Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy. Journal of visualized experiments : JoVE. 2010;(44):2061.
Dale, B., McNerney, G. P., Thompson, D. L., Hübner, W., Huser, T., & Chen, B. K. (2010). Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy. Journal of visualized experiments : JoVE(44), 2061. doi:10.3791/2061
Dale, B., McNerney, G. P., Thompson, D. L., Hübner, W., Huser, T., and Chen, B. K. (2010). Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy. Journal of visualized experiments : JoVE, 2061.
Dale, B., et al., 2010. Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy. Journal of visualized experiments : JoVE, (44), p 2061.
B. Dale, et al., “Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy”, Journal of visualized experiments : JoVE, 2010, pp. 2061.
Dale, B., McNerney, G.P., Thompson, D.L., Hübner, W., Huser, T., Chen, B.K.: Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy. Journal of visualized experiments : JoVE. 2061 (2010).
Dale, Benjamin, McNerney, Gregory P, Thompson, Deanna L, Hübner, Wolfgang, Huser, Thomas, and Chen, Benjamin K. “Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy”. Journal of visualized experiments : JoVE 44 (2010): 2061.

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