Nitric oxide synthase: MRNA expression of different isoforms in human monocytes/macrophages

Reiling, Norbert; Ulmer, Artur J.; Duchrow, M.; Ernst, Martin; Flad, Hans‐Dieter; Hauschildt, Sunna

doi:10.1002/eji.1830240836

Cited by 200 publications

(91 citation statements)

References 24 publications

(21 reference statements)

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“…The production of NO by monocytes in alcoholic liver disease [13], and the stimulation of NO release in vitro by human monocytes, when brought into contact with tumor cells, has also been reported [14]. More recently, expression of iNOS mRNA in human monocytes stimulated with LPS/IFN-Z has been demonstrated [15]. However, doubts regarding the release of NO by human monocytes still remain, since other workers have been unable to detect products related to this nitrogen intermediate in cultures of these cells with cytokines or LPS [VVI6].…”

Section: Discussionmentioning

confidence: 88%

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Inducible nitric oxide synthase in human lymphomononuclear cells activated by synthetic peptides derived from extracellular matrix proteins

et al. 1995

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Synthetic peptides with sequences present in extracellular matrix proteins are capable of causing the expression of the inducible form of nitric oxide synthase (iNOS), detected by immunocytochemistry, and the release of NO by human lymphomononuclear cells incubated in their presence. Active peptides are 15-mers containing a characteristic 2-6-11 motif in which the amino acid residue at position 2 is Leu, Ile, Val, Gly, Ala or Lys; the residue at position 6 is always Pro; and residue 11 is Glu or Asp. The induction of iNOS in human monocytes and macrophages could be involved in the eytotoxicity against tumor cell lines also elicited by these peptides.

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Section: Discussionmentioning

confidence: 88%

“…It is now widely accepted that NO synthesized from L-arginine in a reaction catalyzed by NOS is an antimicrobial and antitumoral effector system of mononuclear cells from rodents [7][8][9]. However, it is still controversial whether an analogous system is also present in human macrophages [10][11][12][13][14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Inducible nitric oxide synthase in human lymphomononuclear cells activated by synthetic peptides derived from extracellular matrix proteins

et al. 1995

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“…Its biological effect in regulating cell survival has been extensively studied (18 eNOS and the neuronal or type I NOS are constitutively expressed (19), and their catalytic activity is regulated by transient calcium increases through the formation of Ca 2+ /calmodulin (CaM) complex (20,21). The presence of eNOS, which was initially cloned from endothelial cells (22,23), has also been detected in human monocytes/ macrophages and subpopulations of T and B lymphocytes (24,25 (14) and mesangial cells (26). eNOS is a calcium-dependent NADPH oxidase with a C-terminal reductase domain that binds NADPH, FAD, and FMN, and an N-terminal oxygenase domain that binds a heme moiety, tetrahydrobiopterin (BH4), and L-arginine (27)(28)(29).…”

Section: Discussionmentioning

confidence: 99%

“…Total RNA isolation and analysis of mRNA expression by real-time RT-PCR or RT-PCR U937 cells were collected at 6,12,18,24,30,36, and 42 h after co-cultured with isogenic H. pylori strains. Total RNA was isolated using Trizol reagent (Life Technologies) according to the manufacturer's instructions.…”

Section: Transfection and Construction Of Stably Transfected Cell Linementioning

confidence: 99%

H. pylori Escape Host Immunoreaction Through Inhibiting ILK Expression by VacA

Yuan

Tao

et al. 2009

Cell Mol Immunol

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Helicobacter pylori (H. pylori) persistently colonizes the gastric mucosa despite a vigorous immune response. Vacuolating cytotoxin secreted by H. pylori has turned out to be a potent immunomodulatory toxin, but the signal transduction pathways involved has not been studied in macrophages. We observed in this study that vacA-deficient H. pylori induced significantly higher expression of integrin-linked kinase (ILK) and endothelial nitric oxygen synthase (eNOS), and significantly more production of reactive oxygen species (

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“…In response to the difficulties in demonstrating NOS production and NO release by human cells, we focused our studies on more sensitive methods to identify NOS in human AEC II by analyzing mRNA expression of NOS2 and NOS3 by RT-PCR (1,28). A critical point of this approach is the purity of the cells, since the high sensitivity of the assay may detect mRNA of contaminating cells.…”

Section: Characteristics Of Isolated Aec IImentioning

confidence: 99%

Pattern of NOS2 and NOS3 mRNA expression in human A549 cells and primary cultured AEC II

Pechkovsky

Zissel

Goldmann

et al. 2002

American Journal of Physiology-Lung Cellular and Molecular Phys

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The human alveolar type II epithelium-like cell line A549 expresses nitric oxide synthase type 2 (NOS2), but not NOS3, and produces nitric oxide (NO) upon appropriate stimulation. However, relatively little is known regarding the NOS2 and NOS3 expression of type II human alveolar epithelial cells (AEC II) in primary culture. We detected NOS3 mRNA in freshly isolated AEC II and after 24 h of culture. NOS3 mRNA levels were much higher in AEC II cultured for 24 h with or without interferon-␥, interleukin-1␤, and tumor necrosis factor-␣, compared with freshly isolated cells. Cytokine stimulation did not change the NOS3 mRNA expression level in AEC II compared with unstimulated cells. NOS3 protein expression was verified by Western blot, and measuring nitrate/nitrite revealed that the protein is active. In contrast, neither NOS2 mRNA nor protein could be detected in freshly isolated, unstimulated or cytokine-stimulated human AEC II in 24-or 72-h primary cultures, whereas A549 cells expressed NOS2 message and protein upon stimulation with proinflammatory cytokines. In situ hybridization confirmed that AEC II express NOS3, but not NOS2 mRNA in vivo. These data demonstrate that there are significant differences between primary AEC II and A549 cells in NOS mRNA expression pattern.nitric oxide synthase; mRNA; RT-PCR; interferon-␥; tumor necrosis factor-␣; interleukin-1␤; type II human alveolar epithelial cells; A549 cell line NITRIC OXIDE SYNTHASES (NOS) constitute a family with at least three distinct isoforms; neuronal (nNOS or NOS1), inducible (iNOS or NOS2) and endothelial (eNOS or NOS3) (20, 21). NOS1 and NOS3 are constitutively active and produce small amounts of nitric oxide (NO), whereas NOS2 is induced by inflammatory cytokines and produces a larger amount of NO. All three types of NOS are expressed in human lung, and NOS mRNA expression and enzyme activity have been described in human bronchial epithelial cells, macrophages, endothelial cells, and vascular smooth muscle cells (18,34). However, relatively little is known regarding the expression of NOS2 and NOS3 in type II human alveolar epithelial cells (AEC II) and how these enzymes are regulated. Although several laboratories have reported that rat AEC II express NOS2 mRNA and produce NO in vitro (12,26) and that human alveolar epithelium exhibits an NOS-like enzymatic activity in vivo (18), there has been neither molecular nor biochemical confirmation of the expression of NOS2 and NOS3 in human primary cultured AEC II. Asano et al. (4) and Kwon and George (19) have demonstrated that the human type II alveolar epithelial cell line (A549) expresses NOS2 and produces NO in response to proinflammatory cytokines such as interferon-␥ (IFN-␥), interleukin-1␤ (IL-1␤), and tumor necrosis factor-␣ (TNF-␣). Therefore, the aims of the present study were 1) to determine whether human AEC II express NOS2 and NOS3 mRNA in primary culture, 2) to characterize the regulation of mRNA expression of NOS isoforms in these cells by proinflammatory cytokines, and 3) to compare these...

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Nitric oxide synthase: MRNA expression of different isoforms in human monocytes/macrophages

Cited by 200 publications

References 24 publications

Inducible nitric oxide synthase in human lymphomononuclear cells activated by synthetic peptides derived from extracellular matrix proteins

Inducible nitric oxide synthase in human lymphomononuclear cells activated by synthetic peptides derived from extracellular matrix proteins

H. pylori Escape Host Immunoreaction Through Inhibiting ILK Expression by VacA

Pattern of NOS2 and NOS3 mRNA expression in human A549 cells and primary cultured AEC II

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