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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:40117*  Cite 
Description

A single computed slice through a tomographic reconstruction of a virological synapse formed between naturally HTLV-1 infected CD4+ T cells and autologous uninfected CD4+ target cells. HTLV-1 particles in the synaptic cleft were identified by immunolabeling using anti-Gag p19 labeling and peroxidase-DAB detection. This image has been downsampled from the raw data image which can be accessed using the link provided to the Cell Centered Database. For more information see: Majorovits et al. (2008) Human T-Lymphotropic Virus-1 visualized at the virological synapse by electron tomography. PLoS ONE 3: e2251, doi:10.1371/journal.pone.0002251, PMID 18509526

Technical Details

CD4+ T cells were isolated and cultured overnight to allow spontaneous expression of HTLV-1 proteins, and subsequently prepared for EM. To detect HTLV-1 Gag matrix protein in infected cells the samples were labelled against HTLV-1 Gag p19 (Tanaka et al., 1986) using the DAB kit from Molecular Probes (Invitrogen Ltd, Paisley, UK ). After the PFA/glutaraldehyde fixation, the samples were incubated in PBS containing 1% BSA, 0.1% Triton X-100 and 2% H2O2, for 1 h at 37uC to block non-specific antibody binding and to quench the endogenous peroxidase activity of the cells. To detect the HTLV-1 complexes, the cells were incubated with 5 mg/ml of anti-Gag p19 (GIN7 mAb) [33] diluted in PBS 1% BSA, for 1 h at 37uC. Then the second antibody, anti-Mouse mAb conjugated to HRP (Invitrogen Ltd, Paisley, UK ), was added at a final dilution of 1 mg/ml, and incubated for 30 minutes at 37uC. The cells were washed three times with PBS containing 1% BSA between each step. For detailed methods of the preparation, refer to the CCDB link provided.

Biological Sources
NCBI Organism Classification
Homo sapiens
HTLV-1 subtype A
Cell Type
mature CD4 single-positive thymocyte
T cell
Biological Context
Biological Process
initiation of viral infection
Attribution
Names
Endre Majorovits
Charles Bangham
Stephen Fuller
Mohamed Nejmeddine
Link
CCDB 3947
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL40117
Archival Resource Key (ARK)
ark:/b7295/w9cil40117
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
transmission electron microscopy (TEM)
Parameters Imaged
electron density
Data Qualifiers
processed data
Sample Preparation
Methods
osmium tetroxide fixed tissue
glutaraldehyde fixed tissue
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 512px ——
Y 512px ——