Optimization of assay conditions for leukotriene B4 synthesis by neutrophils or platelets isolated from peripheral blood of monogastric animals.
Abstract: Neutrophils are involved in inflammation through leukotriene (LT) production. The predominant proinflammatory leukotriene released from neutrophils is LTB4, which serves as a biological marker of inflammation. The purpose of this study was to optimize the conditions ex vivo for LTB4 production by neutrophils from horses and dogs, and platelets from chickens. Optimal production of LTB4 was characterized by incubation time (2.5, 5, 10, 15 or 20 min), temperature (25 or 37 degrees C), and calcium ionophore A23187 concentration (0.1, 1, 10 or 20 microM). Incubation longer than 2.5 min did not increase production of LTB4 in chickens or horses; in dogs, incubation for 2.5 and 10 min resulted in the highest concentrations of LTB4 (P<or=0.05). In all species, incubation at 37 degrees C resulted in optimal LTB4 production (P<or=0.05). Production of LTB4 was highest when neutrophils or platelets were stimulated with 20 microM calcium ionophore in all species (P<or=0.05). These results indicate that neutrophils isolated from horses and dogs, and platelets isolated from chickens, are capable of producing LTB4. Optimum conditions for LTB4 production are similar in all three species.
Publication Date: 2005-04-20 PubMed ID: 15919611DOI: 10.1016/j.plefa.2005.03.001Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research focused on determining the best conditions for the production of a proinflammatory compound, Leukotriene B4 (LTB4), by neutrophils and platelets from dogs, horses and chickens.
Background
- Neutrophils are blood cells that are part of the immune system. These cells produce leukotrienes, specifically Leukotriene B4 (LTB4), as part of the body’s inflammatory response to injury or infection.
- It’s essential to optimize the conditions for LTB4 production in the lab as it can serve as a biological marker for inflammation, useful in both clinical and research settings.
Methods and Procedure
- The study was carried out on neutrophils isolated from the blood of horses and dogs, and platelets from chickens.
- Various conditions were tested to determine the optimal environment for LTB4 production. These conditions included varying incubation times (2.5, 5, 10, 15 or 20 min), temperatures (25 or 37 degrees C), and different concentrations of a substance known as calcium ionophore A23187 (0.1, 1, 10 or 20 microM).
- Calcium ionophore A23187 stimulates the cells to produce LTB4.
Results
- The results indicated that incubation for more than 2.5 minutes did not increase LTB4 production in chickens or horses. For dogs, however, both 2.5 and 10 minutes incubation resulted in the highest LTB4 concentrations.
- A temperature of 37 degrees C was found optimal for LTB4 production across all tested species.
- The highest production of LTB4 was observed when the cells were stimulated with 20 microM of calcium ionophore A23187, again across all species.
Implications
- This research provides valuable information on the ideal conditions for LTB4 production from neutrophils and platelets in these three monogastric animal species. This process facilitates easier and more effective measurement of inflammation in these animals, thus contributing to better health monitoring and management.
Cite This Article
APA
Jha S, Hall JA, Cherian G, Henry LR, Schlipf JW.
(2005).
Optimization of assay conditions for leukotriene B4 synthesis by neutrophils or platelets isolated from peripheral blood of monogastric animals.
Prostaglandins Leukot Essent Fatty Acids, 72(6), 423-430.
https://doi.org/10.1016/j.plefa.2005.03.001 Publication
Researcher Affiliations
- Department of Animal Sciences, College of Agricultural Sciences, Oregon State University, Corvallis, Oregon 97331-4802, USA.
MeSH Terms
- Animals
- Blood Platelets / metabolism
- Calcium / pharmacology
- Cells, Cultured
- Chickens
- Dogs
- Horses
- Ionophores / pharmacology
- Leukotriene B4 / biosynthesis
- Neutrophils / metabolism
- Temperature
- Time Factors
Citations
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