Abstract
Bordetella pertussis, the causative agent of whopping cough, produces an adenylate cyclase toxin (CyaA) that plays a key role in the host colonization by targeting innate immune cells which express CD11b/CD18, the cellular receptor of CyaA. CyaA is also able to invade non-phagocytic cells, via a unique entry pathway consisting in a direct translocation of its catalytic domain across the cytoplasmic membrane of the cells. Within the cells, CyaA is activated by calmodulin to produce high levels of cyclic adenosine monophosphate (cAMP) and alter cellular physiology. In this study, we explored the effects of CyaA toxin on the cellular and molecular structure remodeling of A549 alveolar epithelial cells. Using classical imaging techniques, biochemical and functional tests, as well as advanced cell mechanics method, we quantify the structural and functional consequences of the massive increase of intracellular cyclic AMP induced by the toxin: cell shape rounding associated to adhesion weakening process, actin structure remodeling for the cortical and dense components, increase in cytoskeleton stiffness, and inhibition of migration and repair. We also show that, at low concentrations (0.5 nM), CyaA could significantly impair the migration and wound healing capacities of the intoxicated alveolar epithelial cells. As such concentrations might be reached locally during B. pertussis infection, our results suggest that the CyaA, beyond its major role in disabling innate immune cells, might also contribute to the local alteration of the epithelial barrier of the respiratory tract, a hallmark of pertussis.
Highlights
The adenylate cyclase (CyaA) is a major toxin secreted by Bordetella pertussis, the causative agent of whopping cough
We showed that CyaA can invade A549 alveolar epithelial cells and trigger a significant remodeling of their molecular adhesion systems [16] and, more recently, Hasan et al (2018) analyzed the impact of CyaA on the functional integrity of human bronchial epithelial cells cultured at the air-liquid interface [17]
A549 cells were exposed for 1 hr to various concentrations of CyaA and the F-actin structures were visualized by phalloidin staining and focal adhesions were visualized with anti-phosphotyrosin PY99 antibodies (Fig 1)
Summary
The adenylate cyclase (CyaA) is a major toxin secreted by Bordetella pertussis, the causative agent of whopping cough. This toxin plays a key role in the early stage of colonization of the respiratory tract by Bordetella pertussis. Functional and structural consequences of epithelial cell invasion by B. pertussis adenylate cyclase toxin frm.org The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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