Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Virus Mimetic Poly (I:C)-Primed Airway Exosomes Enter Brain and Induce Mitochondrial Reactive Oxygen Species in Microglia

Version 1 : Received: 13 July 2021 / Approved: 15 July 2021 / Online: 15 July 2021 (11:12:49 CEST)

How to cite: Kulakauskienė, D.; Narauskaitė, D.; Gečys, D.; Juknaitė, O.; Jankauskaitė, L.; Masaitytė, A.; Šventoraitienė, J.; Inokaitis, H.; Miknienė, Z.; Sadauskienė, I.; Balion, Z.; Morkūnienė, R.; Paužienė, N.; Pauža, D.; Jekabsone, A. Virus Mimetic Poly (I:C)-Primed Airway Exosomes Enter Brain and Induce Mitochondrial Reactive Oxygen Species in Microglia. Preprints 2021, 2021070356 (doi: 10.20944/preprints202107.0356.v1). Kulakauskienė, D.; Narauskaitė, D.; Gečys, D.; Juknaitė, O.; Jankauskaitė, L.; Masaitytė, A.; Šventoraitienė, J.; Inokaitis, H.; Miknienė, Z.; Sadauskienė, I.; Balion, Z.; Morkūnienė, R.; Paužienė, N.; Pauža, D.; Jekabsone, A. Virus Mimetic Poly (I:C)-Primed Airway Exosomes Enter Brain and Induce Mitochondrial Reactive Oxygen Species in Microglia. Preprints 2021, 2021070356 (doi: 10.20944/preprints202107.0356.v1).

Abstract

Viral infections induce exosomes containing viral material and inflammatory factors. During respiratory tract infection, exosomes can easily cross the blood-brain barrier and transmit the inflammatory signal to the brain; however, such a hypothesis has no experimental evidence. The study investigated whether exosomes 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. Exosomes 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, exosomes were loaded with Alexa Fluor 555-labelled RNA. Intracellular reactive oxygen species (ROS) were evaluated by DCFDA fluo-rescence and mitochondrial superoxide - by MitoSOX. The exosomes 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 exosomes from airway cells untreated with poly (I:C). The data indicate that virus-primed airway cell exosomes might enter the brain and induce the activation of microglial cells.

Subject Areas

airway cell exosomes; viral infection; microglia; mitochondria; reactive oxygen species

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