Abstract
The packaging of molecular constituents inside extracellular vesicles (EVs) allows them to participate in intercellular communication and the transfer of biological molecules, however the role of EVs during bacterial infection is poorly understood. The goal of this study was to examine the effects of Pseudomonas aeruginosa (P. aeruginosa) infection on the biogenesis and composition of EVs derived from the mouse microglia cell line, BV-2. BV-2 cells were cultured in exosome-free media and infected with 0, 1.3 × 104, or 2.6 × 104 colony forming units per milliliter P. aeruginosa for 72 h. The results indicated that compared with the control group, BV-2 cell viability significantly decreased after P. aeruginosa infection and BV-2-derived EVs concentration decreased significantly in the P. aeruginosa-infected group. P. aeruginosa infection significantly decreased chemokine ligand 4 messenger RNA in BV-2-derived infected EVs, compared with the control group (p ≤ 0.05). This study also revealed that heat shock protein 70 (p ≤ 0.05) and heat shock protein 90β (p ≤ 0.001) levels of expression within EVs increased after P. aeruginosa infection. EV treatment with EVs derived from P. aeruginosa infection reduced cell viability of BV-2 cells. P. aeruginosa infection alters the expression of specific proteins and mRNA in EVs. Our study suggests that P. aeruginosa infection modulates EV biogenesis and composition, which may influence bacterial pathogenesis and infection.
Highlights
Pseudomonas aeruginosa (P. aeruginosa) is a Gram-negative, opportunistic pathogen that contributes to chronic airway infections in cystic fibrosis patients [1]
This study examined the cytokine content packaged within microglia-derived extracellular vesicles (EVs) after P. aeruginosa infection; the findings further supported this phenomenon
The trypan blue exclusion assay was performed to quantitate the numbers of viable cells present in BV-2 cell suspensions after P. aeruginosa infection
Summary
Pseudomonas aeruginosa (P. aeruginosa) is a Gram-negative, opportunistic pathogen that contributes to chronic airway infections in cystic fibrosis patients [1]. Pathogens 2019, 8, 297 implicated as the cause of life-threatening illnesses among immunocompromised individuals and burn victims who reside in healthcare facilities (e.g., hospitals, nursing homes [2], and rehabilitation centers [2]). According to the US Centers for Disease Control and Prevention, more than 6000 healthcare-associated multidrug-resistant P. aeruginosa infections occur annually; approximately 400 of these infections result in death. The bacterium can invade the central nervous system from the inner ear or paranasal sinus region. It can be directly inoculated into the brain during head trauma, neurosurgery, or an invasive diagnostic procedure [3]. Because P. aeruginosa has become increasingly drug resistant, recent studies have dissected how P. aeruginosa disturbs immune cells and their ability to communicate with other cells using extracellular vesicles (EVs) [4,5,6,7]
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