Analysis of the effects of lipopolysaccharide on macrophages: differential phagocytic responses of C3H/HeN and C3H/HeJ macrophages in vitro

Marshall, S T; Rosenstreich, David L.

doi:10.1128/iai.25.1.328-336.1979

Cited by 43 publications

(14 citation statements)

References 12 publications

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“…Antibody-mediated uptake of S-LPS. High concentrations of nonopsonized LPS are toxic to LPS-responsive macrophages (24). This observation was confirmed in this study (data not shown) and limited our ability to study the metabolism of larger amounts of cell-associated R-LPS.…”

Section: Resultssupporting

confidence: 66%

“…(C3H/HeN mice), or from Jackson Laboratories, Bar Harbor, Maine (C3H/HeJ mice), and were stored in the animal facility of the University of Texas Health Science Center, Dallas. The mice were injected intraperitoneally with 2.0 ml of sterile Brewer thioglycolate broth (Difco Laboratories, Detroit, Mich.), and peritoneal cells were harvested 5 days later (24). In each experiment, C3H/HeN and C3H/HeJ mice of comparable ages (within 1 week) were used; all experiments were performed when the mice were 5 to 12 weeks old.…”

Section: Methodsmentioning

confidence: 99%

“…In some experiments, the amount of DNA increased or decreased by as much as 30% over the 48-to 72-h incubation period. A decrease in the amount of cell DNA was assumed to represent cell loss (cytotoxicity was usually seen only when C3H/HeN cells were incubated with higher concentrations of LPS [24], and the calculation for the amount of cell-associated LPS (micrograms of LPS per milligram of cell DNA) was adjusted accordingly. An increase in the amount of cell DNA over time was assumed to represent cell proliferation; when this change occurred during the chase period of an experiment, the amount of cell DNA at the end of the load period was used for calculating the amount of cell-associated LPS throughout the experiment.…”

Section: Methodsmentioning

confidence: 99%

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Uptake and deacylation of bacterial lipopolysaccharides by macrophages from normal and endotoxin-hyporesponsive mice

Munford¹,

Hall²

1985

Infect Immun

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Macrophages are thought to play a central role in the responses of animals to gram-negative bacterial lipopolysaccharides (LPS). Since nothing is known about the metabolism of LPS by these cells, we studied the uptake and deacylation of radiolabeled LPS by thioglycolate-elicited peritoneal macrophages from normal (C3H/HeN) and LPS-hyporesponsive (C3H/HeJ) mice. Macrophages from both kinds of mice took up and deacylated LPS that were added to the culture medium. Opsonization of the LPS with anti-LPS immunoglobulin G antibodies greatly increased LPS uptake; the opsonized LPS also underwent deacylation at rates that were directly related to the amount of cell-associated LPS. An analysis of the fatty acid composition of the cell-associated LPS indicated that the cells have one or more acyloxyacyl hydrolases that remove the non-hydroxylated fatty acids that are normally substituted to the hydroxyl groups of (glucosamine-linked) 3-hydroxytetradecanoate residues in lipid A; we also found evidence for deacylation of 3-hydroxytetradecanoate from the glucosamine backbone. LPS deacylation by macrophages from C3H/HeN and C3H/HeJ mice was qualitatively and quantitatively similar. Nonopsonized LPS are able to stimulate LPS-responsive cells; in these studies we established that animal cells can deacylate nonopsonized LPS, thus raising the possibility that LPS metabolism may play a role in modulating cellular stimulation.

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Section: Resultssupporting

confidence: 66%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Uptake and deacylation of bacterial lipopolysaccharides by macrophages from normal and endotoxin-hyporesponsive mice

Munford¹,

Hall²

1985

Infect Immun

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“…According to Vogel et al (16), LPS induces a profound inhibition of Fc-mediated phagocytosis in LPS-responsive M4 of C3H/HeN mice. Furthermore, low concentrations of LPS stimulate phagocytosis in M4X of nonresponder C3H/HeJ mice.…”

Section: Discussionmentioning

confidence: 99%

Contributions of C1q, bacterial lipopolysaccharide, and porins during attachment and ingestion phases of phagocytosis by murine macrophages

Euteneuer¹,

Störkel²,

Loos³

1986

Infect Immun

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In contrast to the S-form of Salmonella minnesota, its Re mutant binds to mouse peritoneal macrophages. The binding reaction triggers an oxidative burst, measured by a chemiluminescent reaction. The oxidative burst was abolished in the presence of either purified lipopolysaccharide or porins (outer membrane proteins) extracted from the Re mutant, suggesting that both components are involved in binding of the Re mutant to macrophages. In addition, Fc-recognizing membrane structures on the macrophage surface bind the Re mutant. Preincubation of macrophages with the Re mutant abolishes immunoglobulin G-sensitized erythrocyte-induced chemiluminescence. Macrophages preincubated with immunoglobulin G-sensitized erythrocytes had a low chemiluminescent signal, and after treatment of the cells with the Re mutant, there was an additional, higher signal. Binding of purified C1q to the Re mutant decreased the adherence of the Re mutant to macrophages, resulting in a diminished chemiluminescent signal. Blocking of endogenous macrophage membrane-associated C1q with a monoclonal antibody [F(ab')2 fragment] directed against mouse macrophages (recognizes the A and B chains of C1q) diminished the oxidative burst. Therefore, the endogenous C1q of macrophages also appears to be involved in attachment of the S. minnesota Re mutant.

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“…Most of the published studies used extreme pharmacologic concentrations of LPS, ranging between 50 and 1000 mg/mL, which exceed the physiologically achievable concentrations by 10 4to 10 7 -fold, to assess various biological responses. 23e27 At these extreme concentrations, LPS causes rapid cell death in various cell types studied, including intestinal and immune cells, 24,25,27,28 and does not provide accurate depiction of biological activity of LPS as of the immune response in the cells and the activation of many downstream transcription factors. In contrast, we previously showed, for the first time, that LPS at physiologically achievable concentrations (0 to 1000 pg/mL) and clinically achievable concentrations (0 to 10 ng/mL) does not cause acute cell death.…”

mentioning

confidence: 99%

Lipopolysaccharide-Induced Increase in Intestinal Epithelial Tight Permeability Is Mediated by Toll-Like Receptor 4/Myeloid Differentiation Primary Response 88 (MyD88) Activation of Myosin Light Chain Kinase Expression

Nighot

Al–Sadi

Guo

et al. 2017

The American Journal of Pathology

163

123

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Lipopolysaccharides (LPSs) are a major component of the Gram-negative bacterial cell wall and play an important role in mediating intestinal inflammatory responses in inflammatory bowel disease. Although recent studies suggested that physiologically relevant concentrations of LPS (0 to 1 ng/mL) cause an increase in intestinal epithelial tight junction (TJ) permeability, the mechanisms that mediate an LPS-induced increase in intestinal TJ permeability remain unclear. Herein, we show that myosin light chain kinase (MLCK) plays a central role in the LPS-induced increase in TJ permeability. Filter-grown Caco-2 intestinal epithelial monolayers and C57BL/6 mice were used as an in vitro and in vivo intestinal epithelial model system, respectively. LPS caused a dose- and time-dependent increase in MLCK expression and kinase activity in Caco-2 monolayers. The pharmacologic MLCK inhibition and siRNA-induced knock-down of MLCK inhibited the LPS-induced increase in Caco-2 TJ permeability. The LPS increase in TJ permeability was mediated by toll-like receptor 4 (TLR-4)/MyD88 signal-transduction pathway up-regulation of MLCK expression. The LPS-induced increase in mouse intestinal permeability also required an increase in MLCK expression. The LPS-induced increase in intestinal permeability was inhibited in MLCK and TLR-4 mice. These data show, for the first time, that the LPS-induced increase in intestinal permeability was mediated by TLR-4/MyD88 signal-transduction pathway up-regulation of MLCK. Therapeutic targeting of these pathways can prevent an LPS-induced increase in intestinal permeability.

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Analysis of the effects of lipopolysaccharide on macrophages: differential phagocytic responses of C3H/HeN and C3H/HeJ macrophages in vitro

Cited by 43 publications

References 12 publications

Uptake and deacylation of bacterial lipopolysaccharides by macrophages from normal and endotoxin-hyporesponsive mice

Uptake and deacylation of bacterial lipopolysaccharides by macrophages from normal and endotoxin-hyporesponsive mice

Contributions of C1q, bacterial lipopolysaccharide, and porins during attachment and ingestion phases of phagocytosis by murine macrophages

Lipopolysaccharide-Induced Increase in Intestinal Epithelial Tight Permeability Is Mediated by Toll-Like Receptor 4/Myeloid Differentiation Primary Response 88 (MyD88) Activation of Myosin Light Chain Kinase Expression

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