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J Microbiol Immunol Infect ; 56(2): 257-266, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36127231

RESUMO

BACKGROUND: The exploration of virology knowledge was limited by the optical technology for the observation of virus. Previously, a three-dimensional multi-resolution real-time microscope system (3D-MRM) was developed to observe the uptake of HIV-1-tat peptide-modified nanoparticles in cell membrane. In this study, we labeled HIV-1 virus-like particles (VLPs) with passivated giant quantum dots (gQDs) and recorded their interactive trajectories with human Jurkat CD4 cells through 3D-MRM. METHODS: The labeled of gQDs of the HIV-1 VLPs in sucrose-gradient purified viral lysates was first confirmed by Cryo-electronic microscopy and Western blot assay. After the infection with CD4 cells, the gQD-labeled VLPs were visualized and their extracellular and intracellular trajectories were recorded by 3D-MRM. RESULTS: A total of 208 prime trajectories was identified and classified into three distinct patterns: cell-free random diffusion pattern, directional movement pattern and cell-associated movement pattern, with distributions and mean durations were 72.6%/87.6 s, 9.1%/402.7 s and 18.3%/68.7 s, respectively. Further analysis of the spatial-temporal relationship between VLP trajectories and CD4 cells revealed the three stages of interactions: (1) cell-associated (extracellular) diffusion stage, (2) cell membrane surfing stage and (3) intracellular directional movement stage. CONCLUSION: A complete trajectory of HIV-1 VLP interacting with CD4 cells was presented in animation. This encapsulating method could increase the accuracy for the observation of HIV-1-CD4 cell interaction in real time and three dimensions.


Assuntos
Linfócitos T CD4-Positivos , Membrana Celular , HIV-1 , Microscopia Eletrônica , Pontos Quânticos , Produtos do Gene tat do Vírus da Imunodeficiência Humana , Humanos , Linfócitos T CD4-Positivos/fisiologia , Linfócitos T CD4-Positivos/ultraestrutura , Linfócitos T CD4-Positivos/virologia , HIV-1/fisiologia , HIV-1/ultraestrutura , Imageamento Tridimensional/métodos , Produtos do Gene tat do Vírus da Imunodeficiência Humana/fisiologia , Peptídeos Penetradores de Células/fisiologia , Membrana Celular/fisiologia , Membrana Celular/ultraestrutura , Membrana Celular/virologia , Nanopartículas/ultraestrutura , Nanopartículas/virologia , Partículas Artificiais Semelhantes a Vírus/fisiologia , Microscopia Eletrônica/métodos
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