Endotoxin-induced tumor necrosis factor activity production by equine peritoneal macrophages.
Abstract: A study was performed to determine whether equine macrophages produce tumor necrosis factor (TNF) activity in vitro in response to endotoxin and to study the effects of endotoxin concentration and incubation time on the amount of TNF produced. Equine peritoneal macrophages were isolated and cultured in vitro for 2, 6, 12, or 24 hr in tissue culture media containing 1) no additive (nonstimulated control), 2) endotoxin (0.5 ng/ml, 5 ng/ml, or 5 micrograms/ml), or 3) the calcium ionophore A23187 (0.95 microM). The supernatant media concentrations of TNF activity were determined by an in vitro cytotoxicity bioassay using the murine fibrosarcoma cell line WEHI 164 clone 13. At all incubation times, macrophages exposed to endotoxin produced significantly more TNF than did nonstimulated control cells (P less than 0.05). Among the three doses of endotoxin tested, there were no differences in concentrations of TNF activity in the supernatant media. The mean concentration of TNF activity in media from nonstimulated macrophages was significantly less after 24 hr incubation than after 2, 6 or 12 hr. Incubation time had no effect on TNF production by macrophages exposed to 0.5 ng/ml and 5 micrograms/ml endotoxin, but macrophages exposed to 5 ng/ml endotoxin produced significantly more TNF after 6 and 12 hr than after 2 hr. To address the reason for the decrease in TNF activity in media from nonstimulated macrophages over time, a second study was designed to determine the effect of incubation on the activity of TNF under tissue culture conditions. Human recombinant TNF was added to media that was incubated 2-24 hr.(ABSTRACT TRUNCATED AT 250 WORDS)
Publication Date: 1990-03-01 PubMed ID: 2178798
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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The research article investigates if equine (horse) macrophages produce tumor necrosis factor (TNF) when exposed to endotoxins and analyzes how different levels of endotoxins and duration of exposure influence the amount of TNF produced.
Research Methodology
- Peritoneal macrophages cells were isolated from horses and cultured in vitro (outside their usual biological context) with different durations: 2, 6, 12, or 24 hours.
- These cells were exposed to three types of culture media: one without any additives (the control group), one containing endotoxin at different concentrations (0.5 ng/ml, 5 ng/ml, or 5 micrograms/ml), and one with a calcium ionophore (A23187).
- The TNF activity in the supernatant (liquid part of a culture medium) was assessed using cytotoxicity bioassay with a mouse cell line.
Results
- The macrophages exposed to endotoxins produced more TNF compared to the control group. This indicates that endotoxins stimulate TNF production.
- There was no difference in the amount of TNF activity observed between the macrophages exposed to the three different concentrations of endotoxins. This suggests that the level of endotoxin exposure does not significantly influence TNF production.
- The TNF activity in the control group decreased significantly after 24 hours, but there was no difference in TNF levels in macrophages exposed to the lowest and highest levels of endotoxins over time.
- The macrophages exposed to the mid-level endotoxin concentration produced more TNF after 6 and 12 hours compared to after 2 hours.
Secondary Study
- A subsequent investigation was conducted to understand the decline in TNF activity in the control group over time.
- Recombinant human TNF was added to culture media and incubated for varying durations to assess the effect of incubation on TNF activity.
- The findings of this secondary study were not included in the abstract, highlighting that it possibly focuses on the stability of TNF under tissue culture conditions.
Cite This Article
APA
Morris DD, Moore JN, Fischer K, Tarleton RL.
(1990).
Endotoxin-induced tumor necrosis factor activity production by equine peritoneal macrophages.
Circ Shock, 30(3), 229-236.
Publication
Researcher Affiliations
- Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens.
MeSH Terms
- Animals
- Cell Line
- Endotoxins / pharmacology
- Escherichia coli
- Female
- Horses
- Lipopolysaccharides / pharmacology
- Macrophages / drug effects
- Macrophages / metabolism
- Male
- Tumor Necrosis Factor-alpha / biosynthesis
Citations
This article has been cited 5 times.- Rütten S, Schusser GF, Abraham G, Schrödl W. Release kinetics of tumor necrosis factor-α and interleukin-1 receptor antagonist in the equine whole blood. BMC Vet Res 2016 Jun 17;12(1):117.
- Karagianni AE, Kapetanovic R, McGorum BC, Hume DA, Pirie SR. The equine alveolar macrophage: functional and phenotypic comparisons with peritoneal macrophages. Vet Immunol Immunopathol 2013 Oct 1;155(4):219-28.
- Jackman BR, Moore JN, Barton MH, Morris DD. Comparison of the effects of ketoprofen and flunixin meglumine on the in vitro response of equine peripheral blood monocytes to bacterial endotoxin. Can J Vet Res 1994 Apr;58(2):138-43.
- Morris DD, Crowe N, Moore JN. Reduced endotoxin-induced production of tumor necrosis factor activity by equine peritoneal macrophages exposed to the dual inhibitor of arachidonic acid metabolism, SK & F 86002. Can J Vet Res 1992 Apr;56(2):110-4.
- Bryant CE, Ouellette A, Lohmann K, Vandenplas M, Moore JN, Maskell DJ, Farnfield BA. The cellular Toll-like receptor 4 antagonist E5531 can act as an agonist in horse whole blood. Vet Immunol Immunopathol 2007 Apr 15;116(3-4):182-9.
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