scholarly journals Modulation of Porcine β-Defensins 1 and 2 upon Individual and Combined Fusarium Toxin Exposure in a Swine Jejunal Epithelial Cell Line

2013 ◽  
Vol 79 (7) ◽  
pp. 2225-2232 ◽  
Author(s):  
Murphy Lam-Yim Wan ◽  
Chit-Shing Jackson Woo ◽  
Kevin J. Allen ◽  
Paul C. Turner ◽  
Hani El-Nezami
Keyword(s):  
Epithelial Cell ◽  
Mrna Expression ◽  
Cell Line ◽  
Defense Mechanisms ◽  
Fumonisin B1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Epithelial Cell Line ◽  
Content Type ◽  
Antimicrobial Effects ◽  
Food And Feed

ABSTRACTDefensins are small antimicrobial peptides (AMPs) that play an important role in the innate immune system of mammals. Since the effect of mycotoxin contamination of food and feed on the secretion of intestinal AMPs is poorly understood, the aim of this study was to elucidate the individual and combined effects of four commonFusariumtoxins, deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEA), and fumonisin B1 (FB1), on the mRNA expression, protein secretion, and corresponding antimicrobial effects of porcine β-defensins 1 and 2 (pBD-1 and pBD-2) using a porcine jejunal epithelial cell line, IPEC-J2. In general, upregulation of pBD-1 and pBD-2 mRNA expression occurred following exposure toFusariumtoxins, individually and in mixtures (P< 0.05). However, no significant increase in secreted pBD-1 and pBD-2 protein levels was observed, as measured by enzyme-linked immunosorbent assay (ELISA). Supernatants from IPEC-J2 cells exposed to toxins, singly or in combination, however, possessed significantly less antimicrobial activity againstEscherichia colithan untreated supernatants. When single toxins and two-toxin combinations were assessed, toxicity effects were shown to be nonadditive (including synergism, potentiation, and antagonism), suggesting interactive toxin effects when cells are exposed to mycotoxin combinations. The results show thatFusariumtoxins, individually and in mixtures, activate distinct antimicrobial defense mechanisms possessing the potential to alter the intestinal microbiota through diminished antimicrobial effects. Moreover, by evaluating toxin mixtures, this improved understanding of toxin effects will enable more effective risk assessments for common mycotoxin combinations observed in contaminated food and feed.

scholarly journals Differential Response of the Human Renal Proximal Tubular Epithelial Cell Line HK-2 to Shiga Toxin Types 1 and 2

2011 ◽  
Vol 79 (9) ◽  
pp. 3527-3540 ◽  
Author(s):  
Erin K. Lentz ◽  
Dinorah Leyva-Illades ◽  
Moo-Seung Lee ◽  
Rama P. Cherla ◽  
Vernon L. Tesh
Keyword(s):  
Epithelial Cell ◽  
Cell Line ◽  
Shiga Toxin ◽  
Renal Damage ◽  
Diarrheal Disease ◽  
Shigella Dysenteriae ◽  
Parp Cleavage ◽  
Epithelial Cell Line ◽  
Cytotoxic Action ◽  
Content Type

ABSTRACTShiga toxins (Stxs) are expressed by the enteric pathogensShigella dysenteriaeserotype 1 and certain serotypes ofEscherichia coli. Stx-producing bacteria cause bloody diarrhea with the potential to progress to acute renal failure. Stxs are potent protein synthesis inhibitors and are the primary virulence factors responsible for renal damage that may follow diarrheal disease. We explored the use of the immortalized human proximal tubule epithelial cell line HK-2 as anin vitromodel of Stx-induced renal damage. We showed that these cells express abundant membrane Gb3and are differentially susceptible to the cytotoxic action of Stxs, being more sensitive to Shiga toxin type 1 (Stx1) than to Stx2. At early time points (24 h), HK-2 cells were significantly more sensitive to Stxs than Vero cells; however, by 72 h, Vero cell monolayers were completely destroyed while some HK-2 cells survived toxin challenge, suggesting that a subpopulation of HK-2 cells are relatively toxin resistant. Fluorescently labeled Stx1 B subunits localized to both lysosomal and endoplasmic reticulum (ER) compartments in HK-2 cells, suggesting that differences in intracellular trafficking may play a role in susceptibility to Stx-mediated cytotoxicity. Although proinflammatory cytokines were not upregulated by toxin challenge, Stx2 selectively induced the expression of two chemokines, macrophage inflammatory protein-1α (MIP-1α) and MIP-1β. Stx1 and Stx2 differentially activated components of the ER stress response in HK-2 cells. Finally, we demonstrated significant poly(ADP-ribose) polymerase (PARP) cleavage after exposure to Stx1 or Stx2. However, procaspase 3 cleavage was undetectable, suggesting that HK-2 cells may undergo apoptosis in response to Stxs in a caspase 3-independent manner.

Transporter mRNA Expression in a Conditionally Immortalized Rat Small Intestine Epithelial Cell Line (TR-SIE)

2004 ◽  
Vol 19 (4) ◽  
pp. 264-269 ◽  
Author(s):  
Ken-ichi Hosoya ◽  
Masatoshi Tomi ◽  
Megumi Takayama ◽  
Yuko Komokata ◽  
Daisuke Nakai ◽  
...  
Keyword(s):  
Small Intestine ◽  
Epithelial Cell ◽  
Mrna Expression ◽  
Cell Line ◽  
Epithelial Cell Line ◽  
Rat Small Intestine

Zinc transporter mRNA expression in the RWPE-1 human prostate epithelial cell line

BioMetals ◽  
2007 ◽  
Vol 21 (4) ◽  
pp. 405-416 ◽  
Author(s):  
Amy L. Albrecht ◽  
Seema Somji ◽  
Mary Ann Sens ◽  
Donald A. Sens ◽  
Scott H. Garrett
Keyword(s):  
Epithelial Cell ◽  
Mrna Expression ◽  
Cell Line ◽  
Zinc Transporter ◽  
Human Prostate ◽  
Epithelial Cell Line ◽  
Prostate Epithelial Cell ◽  
Human Prostate Epithelial Cell

scholarly journals The Tyrosine Kinase BceF and the Phosphotyrosine Phosphatase BceD of Burkholderia contaminans Are Required for Efficient Invasion and Epithelial Disruption of a Cystic Fibrosis Lung Epithelial Cell Line

2014 ◽  
Vol 83 (2) ◽  
pp. 812-821 ◽  
Author(s):  
Ana S. Ferreira ◽  
Inês N. Silva ◽  
Fábio Fernandes ◽  
Ruth Pilkington ◽  
Máire Callaghan ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Epithelial Cell ◽  
Tyrosine Kinase ◽  
Cell Line ◽  
Parental Strain ◽  
Lung Epithelial Cell ◽  
Epithelial Cell Line ◽  
Content Type ◽  
Burkholderia Contaminans ◽  
Lung Epithelial

Bacterial tyrosine kinases and their cognate protein tyrosine phosphatases are best known for regulating the biosynthesis of polysaccharides. Moreover, their roles in the stress response, DNA metabolism, cell division, and virulence have also been documented. The aim of this study was to investigate the pathogenicity and potential mechanisms of virulence dependent on the tyrosine kinase BceF and phosphotyrosine phosphatase BceD of the cystic fibrosis opportunistic pathogenBurkholderia contaminansIST408. The insertion mutantsbceD::Tp andbceF::Tp showed similar attenuation of adhesion and invasion of the cystic fibrosis lung epithelial cell line CFBE41o- compared to the parental strainB. contaminansIST408. In the absence ofbceDorbceFgenes,B. contaminansalso showed a reduction in the ability to translocate across polarized epithelial cell monolayers, demonstrated by a higher transepithelial electrical resistance, reduced flux of fluorescein isothiocyanate-labeled bovine serum albumin, and higher levels of tight junction proteins ZO-1, occludin, and claudin-1 present in monolayers exposed to these bacterial mutants. Furthermore,bceD::Tp andbceF::Tp mutants induced lower levels of interleukin-6 (IL-6) and IL-8 release than the parental strain. In conclusion, although the mechanisms of pathogenicity dependent on BceD and BceF are not understood, these proteins contribute to the virulence ofBurkholderiaby enhancement of cell attachment and invasion, disruption of epithelial integrity, and modulation of the proinflammatory response.

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