A noncanonical autophagy is involved in the transfer of Plasmodium-microvesicles to astrocytes

Autophagy Pathways in the Genesis of Plasmodium-Derived Microvesicles: A Double-Edged Sword?

“…LAP is an intermediary pathway between autophagy and phagocytosis, involving ATG proteins and the recruitment of LC3-II directly on the single-membrane LAPosome formed around the engulfed microbe, prior to its fusion with lysosomes [ 32 ]. The LAP pathway was also found to be an inducer of astrocyte pro-inflammatory responses that play a major role in the pathogenesis of CM [ 28 ].…”

“…Experiments were performed in agreement with the ethics of animal experimentation, and were approved by the French animal welfare committee “Ministère de l’Agriculture et de la Pêche” n°A 75485. All experiments carried out on CM R and TLR3KO mice were conducted as described in the previous work published in CM S [ 28 ]. Primary cultures of astrocytes and GFP- Pb A (1.49L clone, gift of Dr D. Walliker, Institute of Genetics, Edinburgh, UK) were used as previously described in [ 28 ].…”

“…All experiments carried out on CM R and TLR3KO mice were conducted as described in the previous work published in CM S [ 28 ]. Primary cultures of astrocytes and GFP- Pb A (1.49L clone, gift of Dr D. Walliker, Institute of Genetics, Edinburgh, UK) were used as previously described in [ 28 ].…”

“…Purified astrocytes from CM R and TLR3KO mice were stimulated with GFP- Pb A-iRBCs (10:1) at 37 °C for 6 h and removed, then further incubated for 24 h or 48 h. For confocal microscopy, cells were plated on glass slides, and for TEM, on coverslips in 35 mm glass bottom dishes (No 1.5, MatTek, P35G-1.5-10-C), as previously described [ 28 ].…”

“…Gene expression quantification was performed using RT-qPCR RT-qPCR on 1 × 10 6 cells. Cell treatment and processing were conducted as described by Leleu et al [ 28 ].…”