Virus Mimetic Poly (I:C)-Primed Airway Exosome-like Particles Enter Brain and Induce Inflammatory Cytokines and Mitochondrial Reactive Oxygen Species in Microglia

Simple Summary Upper respiratory tract viral infections are among the most common diseases. The blood-brain barrier protects the brain from direct invasion of pathogens. However, the cells share their content with other cells in small nanovesicles called exosomes that can travel long distances and cross biological barriers. Therefore, virus-infected cell extracellular vesicles (EVs) might transmit inflammatory signals or even viral particles to other cells. If they would carry such signals or particles to the central nervous system, it might cause neuroinflammation. However, the migration and impact of virus-primed airway cell EVs on the brain have not been studied yet. Therefore, the study aimed to track airway EVs from the respiratory tract to the brain and determine how infection-primed particles affect microglia—the cells responsible for immune response in the brain. The study revealed that airway cell EVs enter the brain within an hour and gather in microglia. Interestingly, many airway EVs were found in the hippocampus, the region most affected by Alzheimer’s disease. Moreover, EVs from virus-infected airway cells stimulated reactive oxygen species in microglia and induced other inflammation mediators in the brain. Thus, airway cells indeed might communicate inflammatory information to the brain during viral infection. Abstract Viral infections induce extracellular vesicles (EVs) containing viral material and inflammatory factors. Exosomes can easily cross the blood-brain barrier during respiratory tract infection and transmit the inflammatory signal to the brain; however, such a hypothesis has no experimental evidence. The study investigated whether exosome-like vesicles (ELVs) from virus mimetic poly (I:C)-primed airway cells enter the brain and interact with brain immune cells microglia. Airway cells were isolated from Wistar rats and BALB/c mice; microglial cell cultures—from Wistar rats. ELVs from poly (I:C)-stimulated airway cell culture medium were isolated by precipitation, visualised by transmission electron microscopy, and evaluated by nanoparticle analyser; exosomal markers CD81 and CD9 were determined by ELISA. For in vitro and in vivo tracking, particles were loaded with Alexa Fluor 555-labelled RNA. Intracellular reactive oxygen species (ROS) were evaluated by DCFDA fluorescence and mitochondrial superoxide—by MitoSOX. ELVs from poly (I:C)-primed airway cells entered the brain within an hour after intranasal introduction, were internalised by microglia and induced intracellular and intramitochondrial ROS production. There was no ROS increase in microglial cells was after treatment with ELVs from airway cells untreated with poly (I:C). In addition, poly (I:C)-primed airway cells induced inflammatory cytokine expression in the brain. The data indicate that ELVs secreted by virus-primed airway cells might enter the brain, cause the activation of microglial cells and neuroinflammation.