Visualizing cell-to-cell transfer of HIV using fluorescent clones of HIV and live confocal microscopy.
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.
Division of Infectious Diseases, Department of Medicine, Immunology Institute, Mount Sinai School of Medicine.
Benjamin
B
Dale
Gregory P
GP
McNerney
Deanna L
DL
Thompson
Wolfgang
W
Hübner
Thomas
T
Huser
Benjamin K
BK
Chen
D015496:CD4-Positive T-Lymphocytes / Q000502:physiology / Q000821:virology; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000378:metabolism*; D006678:HIV / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015658:HIV Infections / Q000635:transmission / Q000821:virology*; D006801:Humans; D019169:Jurkat Cells; D018613:Microscopy, Confocal / Q000379:methods*; D018715:Microscopy, Video / Q000379:methods; D014162:Transfection; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000032:analysis / Q000378:metabolism*
D054301:gag Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins