Complement-induced equine neutrophil adhesiveness and aggregation.
Abstract: Equine neutrophils (PMN) were isolated from citrated normal blood by density gradient separation on Ficoll-Hypaque to greater than 96% purity and 98% viability and an average of 3.78 x 10(7) PMN/ml. The agonist C5a des Arg was used in serial dilutions of whole zymosan-activated equine plasma (ZAP) or was partially purified from ZAP by column chromatography. Purified equine PMN exhibited rapid aggregation following incubation with C5a des Arg which was further dependent on the availability of divalent cations, especially Mg++. The microfilament disruptive agent cytochalasin B (5 micrograms/50 microliters) greatly augmented aggregation responses to C5a des Arg. Subaggregating doses of C5a des Arg promoted PMN adhesiveness as assayed on 0.5 x 10 cm borosilicate glass columns containing a 2.0 cm bed of Sephadex G-25. This C5a des Arg-induced increased adhesiveness was inhibitable by prior incubation of the PMN with either non-steroidal (0.065 M phenylbutazone) or steroidal (0.005 M dexamethasone) anti-inflammatory agents. Ultrastructural studies correlated well with functional assays and revealed marked organelle-free lamellipodia formation without PMN-PMN contact at subaggregating doses of the agonist and progressive PMN-PMN contact at aggregating doses. Equine PMN are responsive to C5a des Arg, and induced adhesiveness responses can be manipulated by anti-inflammatory agents.
Publication Date: 1987-05-01 PubMed ID: 3603963DOI: 10.1177/030098588702400308Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research studied the bringing together and stickiness of equine neutrophils – a type of white blood cell in horses, under the influence of the cell stimulating molecule C5a des Arg. The study further examined how these cell behaviours implied by C5a des Arg could be tempered by using anti-inflammatory drugs.
Methodology and Results
- The researchers first isolated equine neutrophils from normal horse blood using a density gradient separation method, achieving a high purity and cell viability.
- The cell stimulating molecule, C5a des Arg, was derived from zymosan-activated equine plasma. Subsequently, both whole and partially purified variants of this plasma were used in subsequent experiments.
- Observations showed that the presence of C5a des Arg prompted an accelerated clumping response from the equine neutrophils. Moreover, the bioavailability of divalent cations, particularly Mg++, was found to further enhance this agglutinating reaction.
- When cytochalasin B, a microfilament disruptive agent, was introduced, it was found to notably boost the neutrophil’s aggregative responses to C5a des Arg.
- Utilising comparative low doses of C5a des Arg demonstrated an enhancing effect on the adhesiveness of neutrophils, as tested via borosilicate glass column assays.
- The increase in stickiness due to C5a des Arg could be diminished through the prior treatment of neutrophils with either non-steroidal (phenylbutazone) or steroidal (dexamethasone) anti-inflammatory agents.
- Ultrastructural examinations revealed the formation of clear lamellipodia (protrusions that support cell movement) with minimal cell-to-cell contact at low dosages of the cell stimulating factor. In contrast, higher dosages led to significantly increased cell-to-cell contact.
Conclusion
- In conclusion, the researchers found that equine neutrophils are responsive to the cell stimulating molecule C5a des Arg, which leads to increased aggregation and adhesiveness of these cells.
- Notably, the study demonstrated that these responses could be mitigated through the application of anti-inflammatory agents.
Cite This Article
APA
Slauson DO, Skrabalak DS, Neilsen NR, Zwahlen RD.
(1987).
Complement-induced equine neutrophil adhesiveness and aggregation.
Vet Pathol, 24(3), 239-249.
https://doi.org/10.1177/030098588702400308 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Calcium / pharmacology
- Cell Adhesion
- Cell Aggregation
- Chromatography, Gel
- Complement C5 / analogs & derivatives
- Complement C5 / immunology
- Complement C5a, des-Arginine
- Cytochalasin B / pharmacology
- Dexamethasone / pharmacology
- Female
- Horses / blood
- Horses / immunology
- Magnesium / pharmacology
- Male
- Microscopy, Electron
- Neutrophils / immunology
- Neutrophils / physiology
- Neutrophils / ultrastructure
- Phenylbutazone / pharmacology
- Zymosan / pharmacology
Citations
This article has been cited 6 times.- Abou-El-Hassan H, Zaraket H. Viral-derived complement inhibitors: current status and potential role in immunomodulation. Exp Biol Med (Maywood) 2017 Feb;242(4):397-410.
- Holden W, Slauson DO, Zwahlen RD, Suyemoto MM, Doré M, Neilsen NR. Alterations in complement-induced shape change and stimulus-specific superoxide anion generation by neonatal calf neutrophils. Inflammation 1989 Dec;13(6):607-20.
- Zwahlen RD, Roth DR, Wyder-Walther M. In vitro aggregation of bovine neonatal neutrophils. A comparative study with adult cattle. Inflammation 1990 Aug;14(4):375-87.
- Bochsler PN, Doré M, Neilsen NR, Slauson DO. A monoclonal-antibody-defined adhesion-related antigen on bovine neutrophils is required for neutrophil aggregation. Inflammation 1990 Oct;14(5):499-508.
- McEwen BJ, Wilcock BP, Eyre P. The effect of leukotriene B4, leukotriene C4, zymosan activated serum, histamine, tabanid extract and N-formyl-methionyl-leucyl-phenylalanine on the in vitro migration of equine eosinophils. Can J Vet Res 1990 Oct;54(4):400-4.
- Bochsler PN, Neilsen NR, Dean DF, Slauson DO. Stimulus-dependent actin polymerization in bovine neutrophils. Inflammation 1992 Aug;16(4):383-92.
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