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Characterizing the Duck NLRP3 Inflammasome
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- Author / Creator
- Lee, Michelle
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Ducks, as the natural reservoir host of influenza A virus (IAV), do not exhibit the same detrimental symptoms when infected by IAV as other susceptible host species, such as chickens and humans. A dysregulated NLRP3 inflammasome response activated by IAV has been linked to severe host outcomes, potentially leading to death. It is not known whether dampening of the NLRP3 inflammasome is a mechanism by which ducks avoid damage due to IAV. Here, I have cloned the duck NLRP3 inflammasome components, assessed their function, and examined their expression in duck tissues following an IAV infection. I cloned and expressed recombinant proteins of the duck NLRP3 inflammasome to examine their interactions in vitro using confocal microscopy and immunoprecipitation. I created expression constructs for NLRP3, Caspase-1, and Interleukin-1beta. I was unable to identify ASC, the adaptor molecule apoptosis-associated speck-like protein containing a CARD, one component of the inflammasome, suggesting that the NLRP3 inflammasome may be incomplete in ducks. To activate the duck NLRP3 inflammasome and examine whether the proteins interacted, I used polyinosinic: polycytidylic acid (poly I:C) and nigericin, a known NLRP3 inflammasome agonist. qPCR and RNA-seq were used to investigate the priming step of the duck NLRP3 inflammasome and how it differed from the priming responses of other species. I found little evidence that the TLR3 transcriptional priming pathway that is activated by poly I:C triggered the downstream NLRP3 inflammasome. This suggests that a lower level of activity is exhibited by the NLRP3 inflammasome during an IAV infection in ducks. This may be a mechanism by which the natural reservoir host of IAV could avoid detrimental damage induced by hyper-inflammation and cytokine storms.
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- Subjects / Keywords
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- Graduation date
- Fall 2021
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- Type of Item
- Thesis
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- Degree
- Master of Science
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- License
- This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.