Downregulation of Tumor Necrosis Factor Expression in the Human Mono-Mac-6 Cell Line by Lipopolysaccharide

Haas, Jürgen; Thiel, Corinna; Blömer, Kathi; Weiß, Elisabeth H.; Riethmüller, Gert; Ziegler‐Heitbrock, H. W. L.

doi:10.1002/jlb.46.1.11

Cited by 41 publications

(18 citation statements)

References 8 publications

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“…Thus, regulatory mechanisms must exist to limit Mbk responses to LPS during infection. The adaptive responses ofMb to LPS that we and others have demonstrated provide a way for regulating cytokine release during endotoxemia (20,(33)(34)(35)(36). In the present studies we have shown that LBP promotes LPSinduced adaptation, as M4 adapted using LPS-LBP com- (37) showed that LPS partial structures could block cell responses to LPS without interfering with CD 14-dependent uptake of LPS.…”

Section: Monocytessupporting

confidence: 59%

Lipopolysaccharide (LPS) recognition in macrophages. Participation of LPS-binding protein and CD14 in LPS-induced adaptation in rabbit peritoneal exudate macrophages.

Mathison¹,

Wolfson²,

Steinemann³

et al. 1993

J. Clin. Invest.

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Exposure of rabbit peritoneal exudate macrophages (PEM) or whole blood to picomolar concentrations ofLPS induces adaptation or hyporesponsiveness to LPS. Because of the importance of plasma LPS-binding protein (LBP) and the macrophage cell membrane protein CD14 in recognition of LPS, we examined the effect of LBP on LPS-induced adaptation in PEM. PEM exposed to LPS in the presence of LBP for 8 h were markedly less responsive to subsequent stimulation by LPS than monocytes/macrophages (M+) adapted in the absence of LBP. LPS-induced expression of TNF was sharply reduced in LBP-LPS-adapted PEM, but in contrast these cells remained fully responsive to Staphylococcus aureus peptidoglycan. We considered that specific hyporesponsiveness in LPS-adapted My or in blood monocytes could be due to decreased expression of CD14 or diminished binding of LBP-LPS complexes to CD14. However, flow cytometry analysis revealed only minimal reduction of CD14 expression or CD14-dependent binding of a fluorescent LPS derivative when normo-and hyporesponsive cells were compared. These results show that complexes of LPS and LBP are more effective than LPS alone in inducing adaptation to LPS, and LPS-induced hyporesponsiveness probably results from changes in cellular elements distinct from CD14 that are involved in either LPS recognition or LPS-specific signal transduction. (J. Clin.

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Section: Monocytessupporting

confidence: 59%

Lipopolysaccharide (LPS) recognition in macrophages. Participation of LPS-binding protein and CD14 in LPS-induced adaptation in rabbit peritoneal exudate macrophages.

Mathison¹,

Wolfson²,

Steinemann³

et al. 1993

J. Clin. Invest.

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“…Studies of endotoxin tolerance induced in vitro (26)(27)(28)(29) and in vivo (24,30,31 ) in animals have shown a decrease in the production of several cytokines by macrophages, including dotoxin-induced synthesis of TNFa, IL-I:, and IL-6 of blood monocytes obtained from humans with the sepsis syndrome was recently reported (32). Another recent study found tolerance to endotoxin-induced synthesis of putative IL-1 bioactivity in monocytes from two children with Hemophilus influenza meningitis (33).…”

Section: Introductionmentioning

confidence: 99%

Tolerance to endotoxin-induced expression of the interleukin-1 beta gene in blood neutrophils of humans with the sepsis syndrome.

McCall¹,

Grosso-Wilmoth²,

LaRue³

et al. 1993

J. Clin. Invest.

180

126

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The induction of genes of host cells stimulated by microbial products such as endotoxin and the tolerance of cells to endotoxin excitation play critical roles in the pathogenesis of microbial-induced acute disseminated inflammation with multiorgan failure (the sepsis syndrome). One gene that is induced in phagocytic cells by endotoxin and that appears to play an essential role in the pathogenesis of the sepsis syndrome is IL-1,8. We report here that blood neutrophils (PMN) of patients with the sepsis syndrome (sepsis PMN) are consistently tolerant to endotoxin-induced expression of the IL-1,8 gene, as determined by decreased synthesis of the IL-113 protein and reductions in IL-1,8 mRNA. This down-regulation of the IL-1#j gene in sepsis PMN occurs concomitant with an upregulation in the constitutive expression of the type 2 IL-1 receptor (IL-1R2). These phenotypic changes do not persist in PMN of patients recovering from the sepsis syndrome. Tolerance has stimulus and response specificity since sepsis PMN tolerant to endotoxin can respond normally to Staphylococcus aureus stimulation of IL-1,B production and they normally secrete elastase. Uninfected patients with severe trauma or shock from causes are not tolerant to endotoxin and tolerance is not limited to patients infected with Gram-negative bacteria. The mechanism responsible for tolerance involves pretranslational events and is not due to loss of the CD14 surface protein, a receptor required for endotoxin induction of IL-1B in PMN. The physiological significance of the tolerance to endotoxin and increased expression of IL-1R2 on sepsis PMN is unknown, but may represent an attempt by the host to protect itself from the deleterious effects of disseminated inflammation. (J. Clin. Invest. 1993. 91:853-861.)

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“…It was suggested to be controlled at the cellular level (Greisman et al, 1966) (Freudenberg and Galanos, 1988). LPS tolerance was then adapted to in vitro treatment of monocyte/macrophage cell lines like Mono Mac 6 or RAW 264.7 (Haas et al, 1989;Virca et al, 1989) and here it was shown that the TNF gene was down-regulated at the mRNA level. While downregulation of TNF is the salient feature of LPS tolerance other products of monocytes may even be upregulated (Zhang and Marrison, 1993;Knopf et al, 1994;West et al, 1997) and this includes anti-inflammatory molecules like IL-10 and IL-1 receptor antagonist Randow et al, 1995).…”

Section: Discussionmentioning

confidence: 84%

“…When stimulated with LPS repeatedly, monocytes/ macrophages lose their ability to respond (Haas et al, 1989;Virca et al, 1989;reviewed by Ziegler-Heitbrock, 1995), a process termed desensitization or tolerance. In monocytes tolerance appears to be mediated by an upregulation of the p50 (NF-B1) gene product, which in the nucleus blocks transactivation of cytokine genes.…”

Section: Introductionmentioning

confidence: 99%

Role of p52 (NF-kappaB2) in LPS Tolerance in a Human B Cell Line

Wedel¹,

Frankenberger²,

Sulski³

et al. 1999

Biological Chemistry

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Cells of the weakly CD14 positive human B cell line RPMI 8226, clone 1, will mobilize NF-B (p50/p65 and p50/p50) proteins and produce TNF mRNA when stimulated with lipopolysaccharide (LPS). When such cells are precultured with a low amount of LPS (50 -250 ng/ ml) for 3 -4 days followed by a secondary stimulation with a high dose of LPS (1 g/ml) then the cytokine expression is strongly reduced, i. e. the cells have become tolerant.Western blot analysis of proteins of the NF-B/rel family demonstrates cytoplasmic p50 and p65 for naive B cells plus a low level of p52. While with tolerance induction the pattern of p50 and p65 proteins remains essentially unchanged, the LPS tolerant 8226 cells show a dramatic increase of both p52 protein and its p100 precursor in the cytosol. This p52 is found strongly upregulated in Western blots of extracts from purified nuclei of tolerant cells. Also, gelshift analysis with the -605 B motif of the human TNF 5 -region shows an additional high mobility complex in LPS tolerant cells -a complex that is supershifted with an anti-p52 antibody.Functional analysis with the -1064 TNF 5 -region in front of the luciferase reporter gene demonstrates that transactivation of the TNF promoter is strongly reduced in tolerant cells. Also, overexpression of p52 will suppress activity of TNF promoter reporter gene constructs.Taken together these data show that tolerance to LPS in the human RPMI 8226 B cell line involves upregulation of the p52 (NF-B2) gene, which appears to be instrumental in the blockade of TNF gene expression.

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Downregulation of Tumor Necrosis Factor Expression in the Human Mono-Mac-6 Cell Line by Lipopolysaccharide

Cited by 41 publications

References 8 publications

Lipopolysaccharide (LPS) recognition in macrophages. Participation of LPS-binding protein and CD14 in LPS-induced adaptation in rabbit peritoneal exudate macrophages.

Lipopolysaccharide (LPS) recognition in macrophages. Participation of LPS-binding protein and CD14 in LPS-induced adaptation in rabbit peritoneal exudate macrophages.

Tolerance to endotoxin-induced expression of the interleukin-1 beta gene in blood neutrophils of humans with the sepsis syndrome.

Role of p52 (NF-kappaB2) in LPS Tolerance in a Human B Cell Line

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