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Vet Microbiol. 2019 Dec;239:108462. doi: 10.1016/j.vetmic.2019.108462. Epub 2019 Oct 18.

Mimicking the passage of avian influenza viruses through the gastrointestinal tract of chickens.

Author information

1
Freie Universität Berlin, Faculty of Veterinary Medicine, Institute of Virology, Robert-von-Ostertag-Str. 7-13, Berlin, 14163, Germany.
2
Freie Universität Berlin, Faculty of Veterinary Medicine, Institute of Virology, Robert-von-Ostertag-Str. 7-13, Berlin, 14163, Germany. Electronic address: mveit@zedat.fu-berlin.de.

Abstract

In contrast to human influenza viruses that replicate in the respiratory tract and are airborne transmitted, avian viruses also replicate in gut epithelial cells and are transmitted via the fecal-oral route. On this route, the virus is exposed to destructive fluids of the digestive tract, which are acidic and contain the proteases pepsin (gizzard) or chymotrypsin and trypsin (intestine). Only the latter enzyme activates virus by cleaving hemagglutinin (HA) into HA1 and HA2 subunits. We mimicked the passage of viruses through the gastrointestinal tract by treating them with digestive fluids from chicken and determined titers and integrity of HA by western-blot. Gizzard fluid completely inactivated virions and degrades HA even at a high dilution, but only if the pH was kept acidic. If the fluid is diluted with neutral buffer (mimicking virus uptake with seawater) particles were more resistant. Virions containing an uncleaved HA were even activated suggesting that gastric juice contains a trypsin-like protease. Undiluted intestinal fluid inactivated particles and destroyed HA, but diluted fluid activated virions. A virus isolated from the duck´s intestine is more tolerant against intestinal fluid compared to fowl plague virus suggesting that the former is better adapted to grow in the intestine. We also demonstrate that influenza viruses replicate to high titers in a novel chicken epithelial gut cell line. While viruses with a monobasic HA cleavage site require addition of trypsin, these cells effectively process HA with a polybasic cleavage site, which could be blocked with an inhibitor of the cellular furin protease.

KEYWORDS:

Acidic pH; Bird intestine; Chymotrypsin; Fecal-Oral route; Hemagglutinin; Inactivation; Influenza virus; Pepsin; Transmission; Trypsin

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