Endotoxin-induced production of interleukin 6 by equine peritoneal macrophages in vitro.
Abstract: A study was performed to determine whether equine peritoneal macrophages produce interleukin 6 (IL-6) in vitro in response to endotoxin. Peritoneal fluid was collected from 14 clinically normal adult horses and was used as the source of peritoneal macrophages. Macrophages from each horse were isolated and cultured separately in vitro in the absence or presence of various concentrations (0.5, 5, 500 ng/ml) of endotoxin (lipopolysaccharide from Escherichia coli 055:B5). Culture medium supernatants were collected after 3, 6, 12, and 24 hours' incubation and were frozen at -70 C until assayed for IL-6 activity. Supernatant IL-6 activity was determined by use of a modified colorimetric assay and the murine hybridoma cell line B 13.29 clone B.9, which is dependent on IL-6 for survival. Results indicated that equine peritoneal macrophages produce IL-6 in vitro and that supernatant medium IL-6 activity was significantly (P less than 0.05) increased by exposure to endotoxin. Significant (P less than 0.05) time and treatment effects on macrophage IL-6 production were apparent. The IL-6 activity peaked at 6 or 12 hours' incubation, then remained high through 24 hours' incubation, regardless of endotoxin exposure. Medium IL-6 activity during 3 and 6 hours' incubation was significantly (P less than 0.05) greater in macrophages exposed to 5 or 500 ng of endotoxin/ml than in those exposed to 0.5 ng of endotoxin/ml; however peak IL-6 activity was similar among all endotoxin concentrations. Endotoxin concentration did not have an effect on medium IL-6 activity from macrophages exposed to endotoxin for 12 or 24 hours.
Publication Date: 1992-08-01 PubMed ID: 1510301
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The study investigates the production of interleukin 6 (IL-6) by equine peritoneal macrophages stimulated by endotoxin in laboratory conditions.
Experiment Setup
- The study utilized peritoneal fluid obtained from 14 healthy adult horses to isolate peritoneal macrophages, a type of immune cell.
- The isolated macrophages were cultured separately and stimulated (or not) using three concentrations of endotoxin, a toxic substance produced by certain bacteria. The concentrations used were 0.5, 5, and 500 ng/ml.
- This cultured peritoneal macrophage cells were then incubated for 3, 6, 12, and 24 hours. The medium supernatants, or fluids, were collected and preserved for IL-6 activity assay.
Testing Process
- The level of IL-6 activity in the supernatants was analyzed using a modified colorimetric assay, a type of test that measures the concentration of a substance in a solution. The test employed the B 13.29 clone B.9 cell line, which needs IL-6 to survive.
Key Findings
- The results showcase that endotoxin causes a significant surge in the IL-6 activity in the supernatant medium, indicating that equine peritoneal macrophages produce IL-6 in vitro when exposed to endotoxin.
- The escalation in IL-6 activity, provoked by the exposure to endotoxin, showed significant time and treatment effects on macrophage IL-6 production.
- It peaked at either 6 or 12 hours of incubation, and continued to stay high until the 24 hours incubation point, regardless of endotoxin exposure.
- Macrophages stimulated by a higher concentration of endotoxin (5 or 500 ng/ml) had significantly greater IL-6 activity during the first 3 and 6 hours of incubation compared to those stimulated by the smaller 0.5 ng endotoxin/ml concentration. However, peak IL-6 activity was the same across all endotoxin concentrations.
- For macrophages exposed for 12 or 24 hours, the varying endotoxin concentration had no effect on medium IL-6 activity.
Cite This Article
APA
Morris DD, Crowe N, Moore JN, Moldawer LL.
(1992).
Endotoxin-induced production of interleukin 6 by equine peritoneal macrophages in vitro.
Am J Vet Res, 53(8), 1298-1301.
Publication
Researcher Affiliations
- Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens 30602-7385.
MeSH Terms
- Animals
- Cells, Cultured
- Endotoxins / toxicity
- Escherichia coli
- Horses
- Interleukin-6 / biosynthesis
- Lipopolysaccharides / toxicity
- Macrophages / metabolism
- Peritoneal Cavity / cytology
Grant Funding
- GM 40586 / NIGMS NIH HHS
Citations
This article has been cited 6 times.- Yassine F, Fostok SF, Al Deen NN, Talhouk RS. Endotoxin Triggers Tumor Initiation Events in Nontumorigenic Breast Epithelial Cells and Enhances Invasion-Related Phenotype in Pretumorigenic and Tumorigenic Breast Epithelial Cells.. Int J Inflam 2021;2021:4666380.
- Pyrgiotakis G, Luu W, Zhang Z, Vaze N, DeLoid G, Rubio L, Graham WAC, Bell DC, Bousfield D, Demokritou P. Development of high throughput, high precision synthesis platforms and characterization methodologies for toxicological studies of nanocellulose.. Cellulose (Lond) 2018 Mar 3;25:2303-2319.
- Zhang Y, Nguyen KC, Lefebvre DE, Shwed PS, Crosthwait J, Bondy GS, Tayabali AF. Critical experimental parameters related to the cytotoxicity of zinc oxide nanoparticles.. J Nanopart Res 2014;16(6):2440.
- 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.
- Riihimäki M, Raine A, Elfman L, Pringle J. Markers of respiratory inflammation in horses in relation to seasonal changes in air quality in a conventional racing stable.. Can J Vet Res 2008 Oct;72(5):432-9.
- Whalen KA, Legault H, Hang C, Hill A, Kasaian M, Donaldson D, Bensch GW, Bensch G, Baker J, Reddy PS, Wood N, Ramarao MK, Ellis DK, Csimma C, McKee C, Clark JD, Ryan J, Dorner AJ, O'Toole M. In vitro allergen challenge of peripheral blood induces differential gene expression in mononuclear cells of asthmatic patients: inhibition of cytosolic phospholipase A2alpha overcomes the asthma-associated response.. Clin Exp Allergy 2008 Oct;38(10):1590-605.
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