Enterotoxigenic Escherichia coli Flagellin Inhibits TNF-Induced NF-κB Activation in Intestinal Epithelial Cells

Wang, Gaochan; Geisbrecht, Brian V.; Rueter, Christian; Hardwidge, Philip R.

doi:10.3390/pathogens6020018

Cited by 7 publications

(5 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…TNF-α is multifunctional in that it promotes cell survival by NF-κB signaling activation or initiates programmed cell death; as such, TNF-α plays a prominent role in the homeostatic functions of the gut [62]. Moreover, anti-TNF-α treatments in humans and mice have been associated with the expansion of Escherichia in the microbiome, suggesting a correlation between Escherichia and reduction in TNF-α [63,64]. Thus, we propose that the HF diet is likely contributing to the increased baseline expression of TNF-α in the intestines of the CON group in this study.…”

Section: Discussionmentioning

confidence: 99%

Probiotics Function as Immunomodulators in the Intestine in C57Bl/6 Male Mice Exposed to Inhaled Diesel Exhaust Particles on a High-Fat Diet

Phillippi

Daniel

Nguyen

et al. 2022

Cells

View full text Add to dashboard Cite

Epidemiological studies reveal a correlation between air pollution exposure and gastrointestinal (GI) diseases, yet few studies have investigated the role of inhaled particulate matter on intestinal integrity in conjunction with a high-fat (HF) diet. Additionally, there is currently limited information on probiotics in mitigating air-pollutant responses in the intestines. Thus, we investigated the hypothesis that exposure to inhaled diesel exhaust particles (DEP) and a HF diet can alter intestinal integrity and inflammation, which can be attenuated with probiotics. 4–6-w-old male C57Bl/6 mice on a HF diet (45% kcal fat) were randomly assigned to be exposed via oropharyngeal aspiration to 35 µg of DEP suspended in 35 µL of 0.9% sterile saline or sterile saline (CON) only twice a week for 4 w. A subset of mice was treated with 0.3 g/day of Winclove Ecologic® barrier probiotics (PRO) in drinking water throughout the duration of the study. Our results show that DEP exposure ± probiotics resulted in increased goblet cells and mucin (MUC)-2 expression, as determined by AB/PAS staining. Immunofluorescent quantification and/or RT-qPCR showed that DEP exposure increases claudin-3, occludin, zona occludens (ZO)-1, matrix metalloproteinase (MMP)-9, and toll-like receptor (TLR)-4, and decreases tumor necrosis factor (TNF)-α and interleukin (IL)-10 expression compared to CON. DEP exposure + probiotics increases expression of claudin-3, occludin, ZO-1, TNF-α, and IL-10 and decreases MMP-9 and TLR-4 compared to CON + PRO in the small intestine. Collectively, these results show that DEP exposure alters intestinal integrity and inflammation in conjunction with a HF diet. Probiotics proved fundamental in understanding the role of the microbiome in protecting and altering inflammatory responses in the intestines following exposure to inhaled DEP.

show abstract

Section: Discussionmentioning

confidence: 99%

Probiotics Function as Immunomodulators in the Intestine in C57Bl/6 Male Mice Exposed to Inhaled Diesel Exhaust Particles on a High-Fat Diet

Phillippi

Daniel

Nguyen

et al. 2022

Cells

View full text Add to dashboard Cite

show abstract

“…These findings suggest that cAMP signaling transduces the inhibitory effect of ETEC on AA uptake in Caco-2 cells. Elevated levels of cAMP activate both PKA-and NF-B (PKA-independent mechanism)-mediated signaling pathways (43,44). To determine the contribution of these signaling pathways to the inhibitory effects of ETEC on AA uptake, we pretreated Caco-2 cells with either H-89 (a PKA inhibitor) or celastrol (an NF-B inhibitor) before exposure of ETEC, followed by determination of AA uptake.…”

Section: Role Of Camp-mediated Nf-b Signaling Pathway In the Inhibitory Effect Of Etec Infection On Aa Uptakementioning

confidence: 99%

“…The LT is structurally and functionally related to cholera toxin, and LT activates adenylate cyclase, which leads to an increase the production of intracellular cAMP (25,26,34). This then leads to activation of downstream targets like protein kinase A (PKA) (26,34) and the nuclear factor--light chain enhancer of activated B cells (NF-B) in intestinal epithelial cells (43,44).…”

Section: Introductionmentioning

confidence: 99%

EnterotoxigenicEscherichia coliheat labile enterotoxin inhibits intestinal ascorbic acid uptake via a cAMP-dependent NF-κB-mediated pathway

Ganapathy

Sabui

Marchant

et al. 2019

American Journal of Physiology-Gastrointestinal and Liver Physi

View full text Add to dashboard Cite

Vitamin C is an antioxidant and acts as a cofactor for many enzymatic reactions. Humans obtain vitamin C from dietary sources via intestinal absorption, a process that involves the sodium-dependent vitamin C transporters-1 and -2 (SVCT1 and SVCT2). Enterotoxigenic Escherichia coli (ETEC) infection impacts intestinal absorption/secretory functions, but nothing is known about its effect on ascorbic acid (AA) uptake. Here we demonstrate that infection of Caco-2 cells with ETEC led to a significant inhibition in intestinal AA uptake. This inhibition was associated with a marked reduction in hSVCT1 and hSVCT2 protein, mRNA, and heterogeneous nuclear RNA (hnRNA) expression levels as well as significant inhibition in the activity of both the SLC23A1 and SLC23A2 promoters. Similarly, exposure of mice to ETEC led to a significant inhibition in intestinal AA uptake and reduction in mSVCT1 and mSVCT2 protein, mRNA, and hnRNA expression levels. Inhibition was caused by the action of heat labile enterotoxin (LT), since infecting Caco-2 cells with LT-deficient ETEC (ΔLT) failed to impact AA uptake. Because LT activates adenylate cyclase, we also examined the effect of dibutyryl-cAMP in AA uptake by Caco-2 cells and observed a significant inhibition. Furthermore, treating the cells with celastrol, a specific NF-κB inhibitor, significantly blocked the inhibition of AA uptake caused by ETEC infection. Together, these data demonstrate that ETEC infection impairs intestinal AA uptake through a cAMP-dependent NF-κB-mediated pathway that regulates both SLC23A1 and SLC23A2 transcription. NEW & NOTEWORTHY Our findings demonstrate that heat-labile enterotoxin produced by enterotoxigenic Escherichia coli inhibits AA uptake in intestinal epithelial cells and mouse intestine. This effect is mediated through transcriptional repression of SLC23A1 (SVCT1) and SLC23A2 (SVCT2) via a cAMP-dependent NF-κB signaling pathway.

show abstract

“…To date, most of the published studies with ClearColi® BL21(DE3) have been restricted to complex medium formulations and shake flask cultures to produce enzymes (Liang et al 2015 ; An et al 2019 ; Jobin et al 2019 ; Song et al 2019 ), antigenic proteins (Martínez-Donato et al 2016 ; González-Miro et al 2017 ; Viranaicken et al 2017 ; Watkins et al 2017b , 2017a ; Abdellrazeq et al 2019 , 2020 ), and therapeutic products (Mamat et al 2015 ; Planesse et al 2015 ; Ueda et al 2016 ; Pooe et al 2017 ; Tomczak et al 2017 ; Wang et al 2017 ; Hunt et al 2019 ; Wilding et al 2019a , 2019b ; López et al 2021 ; Nguyen et al 2021 ). ClearColi® BL21(DE3) has also been used to produce diagnostic biomolecules (Masarapu et al 2017 ; Yoo et al 2017 ; Park et al 2021 ) and in specific functional studies (Moon et al 2018 ; Bong et al 2019 ; Hayashi et al 2019 ; Kobayashi et al 2019 ; Qiao et al 2019 ; Carratalá et al 2021 ; Hsu et al 2021 ; Segovia-Trinidad et al 2021 ), among other uses (Ran et al 2019 ; Sankaran et al 2019 ; Sankaran and del Campo 2019 ; Gnopo et al 2020 ).…”

Section: Introductionmentioning

confidence: 99%

ClearColi as a platform for untagged pneumococcal surface protein A production: cultivation strategy, bioreactor culture, and purification

Cardoso

Paredes

Campani

et al. 2022

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Several studies have searched for new antigens to produce pneumococcal vaccines that are more effective and could provide broader coverage, given the great number of serotypes causing pneumococcal diseases. One of the promising subunit vaccine candidates is untagged recombinant pneumococcal surface protein A (PspA4Pro), obtainable in high quantities using recombinant Escherichia coli as a microbial factory. However, lipopolysaccharides (LPS) present in E. coli cell extracts must be removed, in order to obtain the target protein at the required purity, which makes the downstream process more complex and expensive. Endotoxin-free E. coli strains, which synthesize a nontoxic mutant LPS, may offer a cost-effective alternative way to produce recombinant proteins for application as therapeutics. This paper presents an investigation of PspA4Pro production employing the endotoxin-free recombinant strain ClearColi® BL21(DE3) with different media (defined, auto-induction, and other complex media), temperatures (27, 32, and 37 °C), and inducers. In comparison to conventional E. coli cells in a defined medium, ClearColi presented similar PspA4Pro yields, with lower productivities. Complex medium formulations supplemented with salts favored PspA4Pro yields, titers, and ClearColi growth rates. Induction with isopropyl-β- d -thiogalactopyranoside (0.5 mM) and lactose (2.5 g/L) together in a defined medium at 32 °C, which appeared to be a promising cultivation strategy, was reproduced in 5 L bioreactor culture, leading to a yield of 146.0 mg PspA4Pro/g dry cell weight. After purification, the cell extract generated from ClearColi led to 98% purity PspA4Pro, which maintained secondary structure and biological function. ClearColi is a potential host for industrial recombinant protein production. Key points • ClearColi can produce as much PspA4Pro as conventional E. coli BL21(DE3) cells. • 10.5 g PspA4Pro produced in ClearColi bioreactor culture using a defined medium. • Functional PspA4Pro (98% of purity) was obtained in ClearColi bioreactor culture. Graphical abstract Supplementary Information The online version contains supplementary material available at 10.1007/s00253-022-11758-9.

show abstract

Enterotoxigenic Escherichia coli Flagellin Inhibits TNF-Induced NF-κB Activation in Intestinal Epithelial Cells

Cited by 7 publications

References 22 publications

Probiotics Function as Immunomodulators in the Intestine in C57Bl/6 Male Mice Exposed to Inhaled Diesel Exhaust Particles on a High-Fat Diet

Probiotics Function as Immunomodulators in the Intestine in C57Bl/6 Male Mice Exposed to Inhaled Diesel Exhaust Particles on a High-Fat Diet

EnterotoxigenicEscherichia coliheat labile enterotoxin inhibits intestinal ascorbic acid uptake via a cAMP-dependent NF-κB-mediated pathway

ClearColi as a platform for untagged pneumococcal surface protein A production: cultivation strategy, bioreactor culture, and purification

Contact Info

Product

Resources

About