Differential roles of the type I and II secretion systems for the intracellular ABC141 Acinetobacter baumannii infection, which elicits an atypical hypoxia response in endothelial cells
Acinetobacter baumannii poses a substantial global health threat, causing severe multi-drug-resistant infections in hospitalized patients. Circulating clinical isolates present remarkable diversity, with a proportion capable of establishing a transient intracellular niche suitable for persistence, multiplication, and spread. Yet, it remains unknown which bacterial factors mediate the formation and maintenance of this niche, especially within non-phagocytic cells, nor what host responses are elicited. This work demonstrates that the invasive A. baumannii ABC141 strain does not secrete ammonia in endothelial cells as previously shown for other A. baumannii strains multiplying within macrophages but resides in an acidic vacuole devoid of active lysosomal degradative enzymes. This compartment mediates bacterial egress and infection of neighboring cells, promoting dissemination. Using a Dual-RNAseq approach, we mapped the host and bacterial gene expression during the replicative stage of the infection. An atypical hypoxia cell response was observed without significant induction of the HIF1 pathway, with no metabolic shift or disturbance of mitochondria. Surprisingly, ABC141 efficiently grew in hypoxic conditions in culture and within host cells. In addition, we found a bacterial signature reflective of an adaptation to a nutrient-deprived environment. Our work also highlights a differential role for ABC141 secretion systems, with the T1SS assisting intracellular multiplication and the T2SS required for host cell invasion, implicating for the first time the T2SS in the intracellular lifecycle of invasive ABC141 in endothelial cells. Author summaryAcinetobacter baumannii is a major burden for healthcare facilities worldwide, causing severe infections in hospitalized and intensive care patients that are challenging to treat because of extremely high levels of resistance to most antimicrobials. Traditionally seen as extracellular, recent studies have highlighted that a proportion of clinical isolates currently circulating in clinics can invade and multiply inside epithelial and endothelial cells. Yet, how these strains establish an intracellular protected niche remains unknown. Here, we undertook a detailed characterization of the intracellular compartment enclosing multiplying bacteria, the induced host response, and the bacterial genes that contribute to the adaptation to this intracellular environment. We found that the A. baumannii ABC141 clinical strain is well equipped to resist acidic, low oxygen, and low nutrient environments. Furthermore, we discovered it uses its type II secretion system to invade human endothelial cells, the first step of this intracellular lifecycle. This study opens new avenues of research to help develop new antimicrobial strategies to combat this bacteria when shielded within intracellular compartments in the host.