RESUMO
Autophagy is an evolutionarily ancient and highly conserved eukaryotic mechanism that targets cytoplasmic material for degradation. Autophagic flux involves the formation of autophagosomes and their degradation by lysosomes. The process plays a crucial role in maintaining cellular homeostasis and responds to various environmental conditions. While autophagy had previously been thought to be a non-selective process, it is now clear that it can also selectively target cellular organelles, such as mitochondria (referred to as mitophagy) and/or invading pathogens (referred to as xenophagy). Selective autophagy is characterized by specific substrate recognition and requires distinct cellular adaptor proteins. Here we review xenophagic mechanisms involved in the recognition and autolysosomal or autophagolysosomal degradation of different intracellular bacteria. In this context, we also discuss a recently discovered cellular self-defense pathway, termed mito-xenophagy, which occurs during bacterial infection of dendritic cells and depends on a TNF-α-mediated metabolic switch from oxidative phosphorylation to glycolysis.
Assuntos
Autofagossomos/microbiologia , Autofagia , Bactérias/imunologia , Bactérias/patogenicidade , Lisossomos/microbiologia , Animais , Autofagossomos/metabolismo , Citoplasma/microbiologia , Células Dendríticas/metabolismo , Células Dendríticas/microbiologia , Humanos , Lisossomos/metabolismo , Mitocôndrias/metabolismo , Mitocôndrias/microbiologiaRESUMO
Flow cytometry enables the analysis of cells by labeling them with fluorescent probes. We describe a novel flow cytometric approach permitting reliable analysis of Coxiella (C.) burnetii-infected cells. The method quantifies infection-forming units (IFUs) in a dose-dependent manner and allows for the specific detection of infection/replication-competent coxiella in cell cultures.
Assuntos
Coxiella burnetii/isolamento & purificação , Citometria de Fluxo , Animais , Técnicas de Cultura de Células , Células Cultivadas , Contagem de Colônia Microbiana , Coxiella burnetii/metabolismo , Interações Hospedeiro-Patógeno , CamundongosRESUMO
Chlamydiae are bacterial pathogens that grow in vacuolar inclusions. Dendritic cells (DCs) disintegrate these compartments, thereby eliminating the microbes, through auto/xenophagy, which also promotes chlamydial antigen presentation via MHC I. Here, we show that TNF-α controls this pathway by driving cytosolic phospholipase (cPLA)2-mediated arachidonic acid (AA) production. AA then impairs mitochondrial function, which disturbs the development and integrity of these energy-dependent parasitic inclusions, while a simultaneous metabolic switch towards aerobic glycolysis promotes DC survival. Tubulin deacetylase/autophagy regulator HDAC6 associates with disintegrated inclusions, thereby further disrupting their subcellular localisation and stability. Bacterial remnants are decorated with defective mitochondria, mito-aggresomal structures, and components of the ubiquitin/autophagy machinery before they are degraded via mito-xenophagy. The mechanism depends on cytoprotective HSP25/27, the E3 ubiquitin ligase Parkin and HDAC6 and promotes chlamydial antigen generation for presentation on MHC I. We propose that this novel mito-xenophagic pathway linking innate and adaptive immunity is critical for effective DC-mediated anti-bacterial resistance.