Position: PHD student
research center : IPBS CNRS UMR 5089, toulouse TEAM: Phagocyte architecture and dynamics website: http://www.ipbs.fr/phagocyte-architecture-and-dynamics E-MAIL: [email protected] TEL: +33 561175961 expertiseImmunology ; Cell Biology ; Virology ; HIV ; Macrophages
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project : HIV-1 cell-to-cell transfer from infected T cells to macrophages
Description Project :
- A long standing problem in the fight against HIV-1 is the establishment of viral reservoirs that support persistence of the virus in infected patients. Among these reservoirs, macrophages are a key cell population in the early steps of transmission and dissemination of the virus, but also during disease progression. Macrophages can be infected either with cell-free viral particles, but also via viral transfer from a previously infected cell. This latter mode of infection is more rapid and more efficient, and is thought to play a major role in dissemination of the virus through the organism. Using a model of coculture between macrophages and infected T cells, we show that cell fusion is the most efficient mechanism of transfer toward macrophages. This process is relevant in different types of resident macrophages, including synovial, decidual and placental macrophages. It is mediated by the establishment of a specific adhesion structure between infected T cells and macrophages that we characterize thanks to state of the art microscopy techniques!, We also investigate the role of molecular actors such as integrins, tetraspanins and the remodelling of actin cytoskeleton during Tcell/macrophage fusion. Finally, we evaluate the ability of macrophages to fuse with infected T cells in different contexts of activation and polarization. Altogether, this project aims at giving access to a better understanding of the mechanisms involved in myeloid cell fusion, but also bring crucial information for the role of these cells in HIV-1 pathogenesis.
ANIMAL AND CELLULAR MODELS:
- Human primary cells (macrophages, T cells) and cell lines (T cells)
- Human tissue explants (synovial membranes, decidua, placenta…)
TECHNIQUES AND METHODS:
- Coculture of HIV-infected T cells and target macrophages
- Human macrophages activation and polarization
- Flow cytometry
- siRNA-mediated gene silencing in human primary monocyte-derived macrophages
- High resolution flurescence microscopy (confocal, SIM)
- Fluorescence live cell imaging of HIV-infected cells (spinning disk)
- Electron microscopy (SEM, TEM)
- Imaging flow cytometry (ImageStreamX) analysis of macrophages/T cells conjugates
- Ex vivo tissue culture and histological analysis