ABSTRACT
The formation of long-lived, multicellular clusters is a fundamental step in the physiopathology of many disease-causing bacteria. Experiments on abiotic surfaces suggest that bacterial colonization, including initial cluster formation, requires (1) irreversible adhesion, (2) cell proliferation, and (3) a phenotypic transition. However, here we show that on infection of a polarized MDCK epithelium, Pseudomonas aeruginosa (PA) forms long-lived - i.e., permanent - bacterial clusters without requiring irreversible adhesion, cell proliferation, or a phenotypic transition. By combining experiments and a mathematical model, we reveal that the cluster formation process is mediated by type IV pili (T4P). Furthermore, we unveil how T4P quantitatively operate during adhesion, finding that it is a stochastic process that involves an activation time, requires the retraction of pili, and results in reversible attachment. We explain how such reversible attachment process leads to the formation of permanent bacterial clusters and quantify the cluster growth dynamics.
ABSTRACT
For opportunistic pathogens such as Pseudomonas aeruginosa, the mucosal barrier represents a formidable challenge. Infections develop only in patients with altered epithelial barriers. Here, we showed that P. aeruginosa interacts with a polarized epithelium, adhering almost exclusively at sites of multi-cellular junctions. In these sites, numerous bacteria attach to an extruded apoptotic cell or apoptotic body. This dead cell tropism is independent of the type of cell death, as P. aeruginosa also binds to necrotic cells. We further showed that P. aeruginosa is internalized through efferocytosis, a process in which surrounding epithelial cells engulf and dispose of extruded apoptotic cells. Intracellularly, along with apoptotic cell debris, P. aeruginosa inhabits an efferocytic phagosome that acquires lysosomal features, and is finally killed. We propose that elimination of P. aeruginosa through efferocytosis is part of a host defense mechanism. Our findings could be relevant for the study of cystic fibrosis, which is characterized by an exacerbated number of apoptotic cells and ineffective efferocytosis.