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Human renal epithelial cells express iNOS in response to cytokines but not bacteria.

Poljakovic, Mirjana LU ; Karpman, Diana LU ; Svanborg, Catharina LU and Persson, Katarina LU (2002) In Kidney International 61(2). p.444-455
Abstract
BACKGROUND: Epithelial cells form the mucosal barriers that prevent the entry of mucosal pathogens, and respond to bacterial infections by producing various host defense molecules. In this study, we examined the inducible nitric oxide synthase (iNOS) response of primary human renal tubular epithelial cells (HRTEC) following infection with uropathogenic Escherichia coli Hu734, or stimulation with lipopolysaccharide (LPS) or cytokines. METHODS: Induction of iNOS was examined by RT-PCR, Western blot, immunohistochemistry and nitrite measurements. The effects of endogenously produced nitric oxide (NO), and exogenously applied DETA/NO, SIN-1 and H2O2 on cell viability were analyzed using a respiration assay. RESULTS: HRTEC did not produce NO... (More)
BACKGROUND: Epithelial cells form the mucosal barriers that prevent the entry of mucosal pathogens, and respond to bacterial infections by producing various host defense molecules. In this study, we examined the inducible nitric oxide synthase (iNOS) response of primary human renal tubular epithelial cells (HRTEC) following infection with uropathogenic Escherichia coli Hu734, or stimulation with lipopolysaccharide (LPS) or cytokines. METHODS: Induction of iNOS was examined by RT-PCR, Western blot, immunohistochemistry and nitrite measurements. The effects of endogenously produced nitric oxide (NO), and exogenously applied DETA/NO, SIN-1 and H2O2 on cell viability were analyzed using a respiration assay. RESULTS: HRTEC did not produce NO following infection with E. coli Hu734, LPS alone, or in combination with interferon-gamma (IFN-gamma), even though these agents caused a marked increase in iNOS expression by RAW 264.7, a macrophage cell line. In contrast, iNOS protein and mRNA expression by HRTEC increased after exposure to a cytokine mixture consisting of interleukin (IL)-1beta, tumor necrosis factor-alpha (TNF-alpha) and IFN-gamma. This was due to the combination of IL-1beta and IFN-gamma, but the individual cytokines had no effect. Inducible NOS-expressing cell cultures showed reduced viability, and this effect was inhibited with the NOS inhibitor L-NMMA in RAW 264.7 cells, but not in HRTEC. HRTEC were more sensitive to oxidative stress induced by H2O2 than to nitrogen stress induced by DETA/NO. CONCLUSIONS: We conclude that uropathogenic E. coli that attach to HRTEC fail to directly activate iNOS expression, and that iNOS expression during bacterial infection is more likely to result from stimulation by local cytokines such as IL-1beta and IFN-gamma. (Less)
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RNA, Oxidative Stress/drug effects/physiology, Messenger/analysis, Tumor Necrosis Factor/pharmacology, Urinary Tract Infections/metabolism, Lipopolysaccharides/pharmacology, Kidney Tubules/*cytology, Interleukin-1/pharmacology, Interferon Type II/pharmacology, Hydrogen Peroxide/pharmacology, Human, Enzymologic/immunology, Gene Expression Regulation, Escherichia coli Infections/*metabolism, Cell Line, Cell Survival/drug effects/physiology, Child, Cytokines/pharmacology, Antineoplastic Agents/pharmacology, Epithelial Cells/drug effects/*enzymology/microbiology, Oxidants/pharmacology, Nitrites/metabolism, Nitric-Oxide Synthase/*genetics/metabolism, Nitric Oxide Donors/pharmacology, Nitric Oxide/metabolism, Molsidomine/*analogs & derivatives/pharmacology, Macrophages/cytology/immunology, Mice, Animal
in
Kidney International
volume
61
issue
2
pages
444 - 455
publisher
Nature Publishing Group
external identifiers
  • wos:000173446600008
  • pmid:11849384
  • scopus:0036156606
ISSN
1523-1755
DOI
10.1046/j.1523-1755.2002.00138.x
language
English
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yes
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f3c27746-567a-4d4b-9af9-29a3edb236d6 (old id 105969)
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http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11849384&dopt=Abstract
date added to LUP
2007-07-06 07:16:13
date last changed
2017-01-01 07:03:21
@article{f3c27746-567a-4d4b-9af9-29a3edb236d6,
  abstract     = {BACKGROUND: Epithelial cells form the mucosal barriers that prevent the entry of mucosal pathogens, and respond to bacterial infections by producing various host defense molecules. In this study, we examined the inducible nitric oxide synthase (iNOS) response of primary human renal tubular epithelial cells (HRTEC) following infection with uropathogenic Escherichia coli Hu734, or stimulation with lipopolysaccharide (LPS) or cytokines. METHODS: Induction of iNOS was examined by RT-PCR, Western blot, immunohistochemistry and nitrite measurements. The effects of endogenously produced nitric oxide (NO), and exogenously applied DETA/NO, SIN-1 and H2O2 on cell viability were analyzed using a respiration assay. RESULTS: HRTEC did not produce NO following infection with E. coli Hu734, LPS alone, or in combination with interferon-gamma (IFN-gamma), even though these agents caused a marked increase in iNOS expression by RAW 264.7, a macrophage cell line. In contrast, iNOS protein and mRNA expression by HRTEC increased after exposure to a cytokine mixture consisting of interleukin (IL)-1beta, tumor necrosis factor-alpha (TNF-alpha) and IFN-gamma. This was due to the combination of IL-1beta and IFN-gamma, but the individual cytokines had no effect. Inducible NOS-expressing cell cultures showed reduced viability, and this effect was inhibited with the NOS inhibitor L-NMMA in RAW 264.7 cells, but not in HRTEC. HRTEC were more sensitive to oxidative stress induced by H2O2 than to nitrogen stress induced by DETA/NO. CONCLUSIONS: We conclude that uropathogenic E. coli that attach to HRTEC fail to directly activate iNOS expression, and that iNOS expression during bacterial infection is more likely to result from stimulation by local cytokines such as IL-1beta and IFN-gamma.},
  author       = {Poljakovic, Mirjana and Karpman, Diana and Svanborg, Catharina and Persson, Katarina},
  issn         = {1523-1755},
  keyword      = {RNA,Oxidative Stress/drug effects/physiology,Messenger/analysis,Tumor Necrosis Factor/pharmacology,Urinary Tract Infections/metabolism,Lipopolysaccharides/pharmacology,Kidney Tubules/*cytology,Interleukin-1/pharmacology,Interferon Type II/pharmacology,Hydrogen Peroxide/pharmacology,Human,Enzymologic/immunology,Gene Expression Regulation,Escherichia coli Infections/*metabolism,Cell Line,Cell Survival/drug effects/physiology,Child,Cytokines/pharmacology,Antineoplastic Agents/pharmacology,Epithelial Cells/drug effects/*enzymology/microbiology,Oxidants/pharmacology,Nitrites/metabolism,Nitric-Oxide Synthase/*genetics/metabolism,Nitric Oxide Donors/pharmacology,Nitric Oxide/metabolism,Molsidomine/*analogs & derivatives/pharmacology,Macrophages/cytology/immunology,Mice,Animal},
  language     = {eng},
  number       = {2},
  pages        = {444--455},
  publisher    = {Nature Publishing Group},
  series       = {Kidney International},
  title        = {Human renal epithelial cells express iNOS in response to cytokines but not bacteria.},
  url          = {http://dx.doi.org/10.1046/j.1523-1755.2002.00138.x},
  volume       = {61},
  year         = {2002},
}