Prado, MonicaMonicaPradoEickel, NinaNinaEickelDe Niz Hidalgo, Mariana IsabelMariana IsabelDe Niz HidalgoHeitmann, AnnaAnnaHeitmannAgop Nersesian, CarolinaCarolinaAgop NersesianWacker, Rahel CorinaRahel CorinaWackerSchmuckli-Maurer, JacquelineJacquelineSchmuckli-MaurerCaldelari, RetoRetoCaldelariJanse, Chris JChris JJanseKhan, Shahid MShahid MKhanMay, JürgenJürgenMayMeyer, Christian GChristian GMeyerHeussler, VolkerVolkerHeussler0000-0001-8028-98252024-10-232024-10-232015-07-24https://boris-portal.unibe.ch/handle/20.500.12422/134659Plasmodium parasites are transmitted by Anopheles mosquitoes to the mammalian host and actively infect hepatocytes after passive transport in the bloodstream to the liver. In their target host hepatocyte, parasites reside within a parasitophorous vacuole (PV). In the present study it was shown that the parasitophorous vacuole membrane (PVM) can be targeted by autophagy marker proteins LC3, ubiquitin, and SQSTM1/p62 as well as by lysosomes in a process resembling selective autophagy. The dynamics of autophagy marker proteins in individual Plasmodium berghei-infected hepatocytes were followed by live imaging throughout the entire development of the parasite in the liver. Although the host cell very efficiently recognized the invading parasite in its vacuole, the majority of parasites survived this initial attack. Successful parasite development correlated with the gradual loss of all analyzed autophagy marker proteins and associated lysosomes from the PVM. However, other autophagic events like nonselective canonical autophagy in the host cell continued. This was indicated as LC3, although not labeling the PVM anymore, still localized to autophagosomes in the infected host cell. It appears that growing parasites even benefit from this form of nonselective host cell autophagy as an additional source of nutrients, as in host cells deficient for autophagy, parasite growth was retarded and could partly be rescued by the supply of additional amino acid in the medium. Importantly, mouse infections with P. berghei sporozoites confirmed LC3 dynamics, the positive effect of autophagy activation on parasite growth, and negative effects upon autophagy inhibition.enATGautophagy related Atg8(yeast) autophagy-related 8 CLSconfocal line scanning CSPcircumsporozoite protein CTSDcathepsin D Cytosolic immune responseLC3 DAPI4′6-diamidino-2-phenylindole DMEMDulbecco modified Eagle medium EBSSEarle balanced salt solution ECACCEuropean Collection of Cell Cultures EMelectron microscopy Exp1exported protein 1 FACSfluorescent-activated cell sorting FCSfetal calf serum GFPgreen fluorescent protein H&Ehematoxylin and eosin HepG2human hepatoma cells IFAimmunofluorescence assay LAMP1lysosomal-associated membrane protein 1 LAPLC3-associated phagocytosis LGALSgalectins MAP1LC3/LC3microtubule-associated protein 1 light chain 3 MEFsmouse embryonic fibroblasts MEMminimum essential medium MTORmechanistic target of rapamycin (serine/threonine kinase) PBSphosphate-buffered saline PEphosphatidylethanolamine PMparasite membrane PVparasitophorous vacuole PVMparasitophorous vacuole membrane PbPlasmodium berghei PbmCherryPlasmodium berghei expressing mCherry fluorescent protein Plasmodium liver-stage PtdIns3Pphosphatidylinositol-3-phosphate RFPred fluorescence protein SDstandard deviation SQSTM1sequestosome 1 STEDstimulated emission depletion UIS4upregulated in infectious sporozoites gene 4 WTwild type atg5−/−autophagy-related 5 knockout e-schzearly schizont hpihours postinfection l-schzlate schizont long term live imaging malaria selective autophagy spzsporozoite ubiquitin500 - Science::570 - Life sciences; biology600 - Technology::630 - Agriculture500 - Sciencearticle10.7892/boris.710172620877810.1080/15548627.2015.1067361