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- Lindblad, Anna, 1987-, et al.
(författare)
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IL-1RA is part of the inflammasome-regulated immune response in bladder epithelial cells and influences colonization of uropathogenic E. coli
- 2019
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Ingår i: Cytokine. - : Academic Press. - 1043-4666 .- 1096-0023. ; 123
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Tidskriftsartikel (refereegranskat)abstract
- The NLRP3 inflammasome, IL-1β release and pyroptosis (cell lysis) have recently been proposed to be essential for the progression of urinary tract infection (UTI) and elimination of intracellular bacterial niches. However, the effects of IL-1R antagonist (IL-1RA) on immune responses during UTI, except for its ability to disrupt IL-1β signalling, are not well understood. The aim of this study was to investigate the role of IL-1RA in UPEC colonization of bladder epithelial cells and the subsequent host inflammatory response. Human bladder epithelial cells (5637) and CRISPR/Cas9 generated NLRP3 and caspase-1 knockdown cells and IL-1RA knockout cells were stimulated with the UPEC isolate CFT073. The results showed that the UPEC virulence factor α-hemolysin is essential for IL-1RA release, and that the inflammasome-associated proteins caspase-1 and NLRP3 affect the release of IL-1RA. IL-1RA deficient cells showed a reduced adherence and invasion by CFT073 compared to wild-type cells, suggesting that IL-1RA may oppose mechanisms that protects against bacterial colonization. A targeted protein analysis of inflammation-related proteins showed that the basal expression of 23 proteins and the UPEC-induced expression of 10 proteins were significantly altered in IL-1RA deficient bladder epithelial cells compared to Cas9 control cells. This suggests that IL-1RA has a broad effect on the inflammatory response in bladder epithelial cells.
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| 3. |
- Lindblad, Anna, 1987-, et al.
(författare)
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The role of caspase-1, caspase-4 and NLRP3 in regulating the host cell response evoked by uropathogenic Escherichia coli
- 2022
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Ingår i: Scientific Reports. - : Nature Publishing Group. - 2045-2322. ; 12:1
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Tidskriftsartikel (refereegranskat)abstract
- The inflammasome-associated proteins caspase-1, caspase-4 and NLRP3 have been emphasised to be essential in the host cell response during urinary tract infection (UTI) by regulating IL-1β release. Our aim was to investigate how the inflammasome-associated proteins regulate the cell response of bladder epithelial cells during infection with uropathogenic Escherichia coli (UPEC). Human bladder epithelial cells (5637) and CRISPR/Cas9 generated caspase-1, caspase-4 and NLRP3 knockdown cells were stimulated with the UPEC strain CFT073. Using Olink proteomics and real time RT-PCR, we showed that caspase-1, caspase-4 and NLRP3 are vital for the expression of many inflammatory genes and proteins from bladder epithelial cells. When investigating the effect of inflammasome-associated proteins on neutrophils, we found that conditioned medium from UPEC-infected caspase-4 knockdown cells significantly increased phagocytosis of CFT073 and significantly decreased ROS production from neutrophils. In contrast, conditioned medium from UPEC-infected NLRP3 knockdown cells significantly decreased the phagocytosis of CFT073 and significantly increased the ROS production from neutrophils. In conclusion, we showed that the inflammasome-associated proteins contribute to the host cell response during UPEC infection.
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| 4. |
- Lindblad, Anna, 1987-
(författare)
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The role of caspase-1, caspase-4, NLRP3 and IL-1RA in bladder epithelial cells infected by uropathogenic Escherichia coli
- 2023
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Urinary tract infection is one of the most common infections and is mostlycaused by uropathogenic Escherichia coli (UPEC). The inflammasomeassociatedproteins caspase-1, caspase-4 and NLRP3 are essential in the hostcell response during urinary tract infection by regulating IL-1β release. Thepro-inflammatory effects of IL-1β can be inhibited by binding of the IL-1receptor antagonist (IL-1RA) to the IL-1 receptor. The aim of this thesis is toinvestigate what role caspase-1, caspase-4, NLRP3 and IL-1RA have on the proinflammatoryhost response evoked by UPEC and their role in recurrent UTI.The results showed that the inflammasome-associated proteinscaspase-1, caspase-4 and NLRP3 are involved in cytokine and chemokinerelease and in antimicrobial activities of neutrophils during UTI. Weconclude that IL-1RA influences the release of various inflammatoryproteins during a UPEC infection from bladder epithelial cells. In addition,deficiency in IL-1RA led to decreased UPEC colonization and invasion ofbladder epithelial cells. Our results also show that NLRP3 has a regulativefunction on estrogen signalling and the expression of antimicrobialpeptides. Additionally, we found that capsase-1 and caspase-4 can regulatethe gene expression of important immune regulators, including TLR4,antimicrobial peptides, cytokines and chemokines.Together, our results show that that the inflammasome-associatedproteins caspase-1, caspase-4, NLRP3 and IL-IRA are important immuneregulatorsduring UPEC infection in bladder epithelial cells. They regulateUPEC colonization, cytokines and chemokines release, antimicrobialactivities of neutrophils and estrogen signalling.
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| 5. |
- Lindblad, Anna, 1987-, et al.
(författare)
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The Role of NLRP3 in Regulation of Antimicrobial Peptides and Estrogen Signaling in UPEC-Infected Bladder Epithelial Cells
- 2023
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Ingår i: Cells. - : MDPI. - 2073-4409. ; 12:18
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Tidskriftsartikel (refereegranskat)abstract
- The NLRP3 inflammasome, estrogen and antimicrobial peptides have all been found to have a vital role in the protection of the bladder urothelium. However, the interdependence between these protective factors during a bladder infection is currently unknown. Our aim was to investigate the role of NLRP3 in the regulation of antimicrobial peptides and estrogen signaling in bladder epithelial cells during a UPEC infection. Human bladder epithelial cells and CRISPR/Cas9-generated NLRP3-deficient cells were stimulated with the UPEC strain CFT073 and estradiol. The gene and protein expression were evaluated with microarray, qRT-PCR, western blot and ELISA. Microarray results showed that the expression of most antimicrobial peptides was reduced in CFT073-infected NLRP3-deficient cells compared to Cas9 control cells. Conditioned medium from NLRP3-deficient cells also lost the ability to suppress CFT073 growth. Moreover, NLRP3-deficient cells had lower basal release of Beta-defensin-1, Beta-defensin-2 and RNase7. The ability of estradiol to induce an increased expression of antimicrobial peptides was also abrogated in NLRP3-deficient cells. The decreased antimicrobial peptide expression might be linked to the observed reduced expression and activity of estradiol receptor beta in NLRP3-deficient cells. This study suggests that NLRP3 may regulate the release and expression of antimicrobial peptides and affect estrogen signaling in bladder epithelial cells.
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