Quantitative 3D Video Microscopy of HIV Transfer Across T Cell Virological Synapses

Hübner, Wolfgang; McNerney, Gregory P.; Chen, Ping; Dale, Benjamin M.; Gordon, Ronald E.; Chuang, Frank Y.S.; Li, Xiaodong; Asmuth, David M.; Huser, Thomas; Chen, Benjamin

doi:10.1126/science.1167525

Cited by 447 publications

(622 citation statements)

References 26 publications

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“…These two modes of adhesion-induced transmission can both contribute to efficient virus cell-to-cell transmission. The here observed behavior of HIV Gag in the context of an MLV Env/receptor synapse is consistent with the observation of lateral movement of HIV to the virological synapse in T cells (13,41). The ability of HIV to utilize surface movement may also explain independence from the cytoplasmic tail of the envelope, although a dependence has also been reported (6,10).…”

Section: Discussionsupporting

confidence: 72%

“…In the case of the human immunodeficiency virus (HIV) and the human T cell lymphoma virus these cell-cell contacts share features with immunological synapses and have been designated infectious or virological synapses (13,14,19,29,41). Using the murine leukemia virus (MLV) as a model system, we demonstrated that the interaction between the viral Env glycoprotein (Env), expressed on the infected cell and the viral receptor, expressed on the uninfected target cell, leads to the establishment of cell-cell contacts (32,45).…”

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confidence: 99%

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Viral Determinants of Polarized Assembly for the Murine Leukemia Virus

Jin

Mothes

2011

J Virol

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Retrovirus transmission via direct cell-cell contact is more efficient than diffusion through the extracellular milieu. This is believed to be due to the ability of viruses to efficiently coordinate several steps of the retroviral life cycle at cell-cell contact sites (D. Viruses exploit and manipulate cell-cell contacts for efficient spreading (12,18,32,37,43). In the case of the human immunodeficiency virus (HIV) and the human T cell lymphoma virus these cell-cell contacts share features with immunological synapses and have been designated infectious or virological synapses (13,14,19,29,41). Using the murine leukemia virus (MLV) as a model system, we demonstrated that the interaction between the viral Env glycoprotein (Env), expressed on the infected cell and the viral receptor, expressed on the uninfected target cell, leads to the establishment of cell-cell contacts (32, 45). Importantly, following the establishment of cellcell adhesion, assembly was polarized toward the cell-cell contact (16). These data suggested that the infected cell can "sense" the uninfected cell and direct the viral components needed to assemble viruses toward the uninfected cell. Sensing of the target cell turned out to be mediated by the accumulation of Env and receptor at the cell-cell interface and depending on the presence of a cytoplasmic tail (C-tail) of Env, MLV Gag was recruited toward the site of cell-cell contact. Here, we identify a tyrosine residue within the C-tail of Env and matrix within Gag as the viral determinants responsible for this process.C MATERIALS AND METHODSPlasmids and reagents. Mutant MLV envelope expression vectors were generated by site-directed mutagenesis based on the Friend MLV envelope expression vector previously described (16,46). MLV Gag-GFP expression vector was made by fusing enhanced green fluorescent protein (eGFP) to C-terminal of Gag and deleting a large fragment of Pol in full-length Friend57 MLV expression vector pLRB303 (23,35). Individual domains of MLV Gag-GFP were deleted or replaced by a standard overlapping PCR approach. MLV Gag-YFP expression vector was previously described (46). Chimeric MLV Gag with MA swapped with HIV MA was kindly provided by Marc Johnson, University of Missouri. HIV Gag-GFP expression vector and chimeric HIV Gag with MA swapped with MLV MA were described previously (17). HEK293 and XC cells were previously described (16).Visualizing virus cell-to-cell transmission. HEK293 cells were transfected with indicated plasmids by FuGENE 6 and cocultured 4 h posttransfection with XC cells expressing mCherry or cyan fluorescent protein (CFP)-tagged mCAT-1 as previously described (16). The formation of MLV synapses was reproducibly observed 2 to 6 postinitiation of cocultures for MLV generated by the tripartite system (MLV GagPol, Gag-YFP, MLV LTR, and MLV Env) or 4 to 10 h postinitiation of cocultures for MLV generated using proviral pLRB303-derived constructs. Imaging of fixed samples, as well as live-cell imaging, was performed using spinning disc confocal microscopy as ...

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Section: Discussionsupporting

confidence: 72%

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confidence: 99%

Viral Determinants of Polarized Assembly for the Murine Leukemia Virus

Jin

Mothes

2011

J Virol

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“…E.g. it is now well known that the human immunodeficiency virus (HIV) can spread very efficiently from an infected T cell to an uninfected T cell through direct cell-cell contact [1][2][3][4]. Imaging this process in living cells is, however, very tedious and time-consuming due to the fact that T cells are non-adherent and only a fraction of cells will contain the virus [4,5].…”

Section: Biophotonicsmentioning

confidence: 99%

Rapid 3D fluorescence imaging of individual optically trapped living immune cells

Wolfson

Steck

Persson

et al. 2014

Journal of Biophotonics

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We demonstrate an approach to rapidly characterize living suspension cells in 4 dimensions while they are immobilized and manipulated within optical traps. A single, high numerical aperture objective lens is used to separate the imaging plane from the trapping plane. This facilitates full control over the position and orientation of multiple trapped cells using a spatial light modulator, including directed motion and object rotation, while also allowing rapid 4D imaging. This system is particularly useful in the handling and investigation of the behavior of non‐adherent immune cells. We demonstrate these capabilities by imaging and manipulating living, fluorescently stained Jurkat T cells. (© 2015 WILEY‐VCH Verlag GmbH &Co. KGaA, Weinheim)

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“…In a separate report in this issue of JASN, Rosolowsky et al 8 did find a secular trend in a similar cohort of patients with type 1 diabetes from the Joslin Clinic. They found a rate of ESRD that was virtually identical to that of the FinnDiane cohort and likewise observed that progression to ESRD was the predominant outcome among their patients with type 1 diabetes and macroalbuminuria.…”

mentioning

confidence: 98%

“…7,8 Cell-to-cell transmission is described mostly between infected T cells or virus-laden antigen-presenting cells and uninfected T cells. This interaction at the site of cell-to-cell contact is called a virologic synapse with polarized T cells forming a protrusion called uropod, which mediates contact with the other cell.…”

mentioning

confidence: 99%