Modulation of an adhesion-related surface antigen on equine neutrophils by bacterial lipopolysaccharide and antiinflammatory drugs.
Abstract: The essential role of the CD11/CD18 family of leukocyte adhesion molecules (LeuCams) in neutrophil-substrate adhesion is well documented. We have found that a monoclonal antibody designated 60.3 (MoAb 60.3) that recognizes the common beta-subunit (CD18) on human neutrophils (PMN) also recognizes a surface antigen on equine PMN. Antigen expression as assessed by immunofluorescence flow cytometry was enhanced by zymosan-activated serum (ZAS) or phorbol 12-myristate 13-acetate (PMA) stimulation. Pretreatment of equine PMN with MoAb 60.3 inhibited ZAS-stimulated aggregation, indicating that the monoclonal recognized a functional epitope on equine PMN involved in adhesion-related functions. Cells pretreated only with bacterial lipopolysaccharide (LPS; 1 microgram/ml) exhibited moderate increased binding of MoAb 60.3 as determined by fluorescence intensity. Preincubation of PMN with LPS resulted in a slight increase in MoAb 60.3 binding after subsequent ZAS stimulation, greater than that with either LPS or ZAS as sole stimulus. Similarly, enhanced binding of MoAb 60.3 was observed with LPS preincubation when PMA was used as a stimulus, but this effect was dose dependent and was observed at only one of three PMA concentrations tested (1 ng/ml). In other experiments, preincubation of PMN with antiinflammatory drugs inhibited 41.5-45.1% of ZAS-stimulated PMN adhesion to monolayers of equine endothelial cells. To determine whether modulation of expression of the adhesion-related antigen recognized by MoAb 60.3 correlated with these observed adhesive responses of PMN, we used immunofluorescence flow cytometry to assess expression of the antigen on drug-treated PMN. Using 10% ZAS as a stimulus, phenylbutazone (PBZ; 100 micrograms/ml) pretreatment of PMN reduced subsequent MoAb 60.3 binding by only 12.3%, and dexamethasone (DEX; 10(-5) M) reduced binding by only 1.0%; reductions of 16.4% with PBZ and 9.3% with DEX occurred when PMA (10 ng/ml) was used as the PMN stimulant. These data suggest that equine PMN express a functional adhesion molecule similar to those found on human PMN and that LPS may enhance the expression of this surface antigen. Expression of this adhesion-related surface antigen on equine PMN does not correlate well with levels of drug-induced diminished adhesion of PMN to endothelium in vitro.
Publication Date: 1990-10-01 PubMed ID: 2394944DOI: 10.1002/jlb.48.4.306Google Scholar: Lookup
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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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This research examines how a specific cell surface antigen, related to cell adhesion, on horse immune cells is altered by bacterial components and anti-inflammatory drugs.
Objectives and Methods
- The research is centered around the CD11/CD18 family of leukocyte adhesion molecules (LeuCams) which are known to play an essential part in the adhesion of neutrophils, a type of white blood cell, to various surfaces.
- A monoclonal antibody named MoAb 60.3, known to recognize the common beta-subunit (CD18) on human neutrophils, has been observed to recognize a similar antigen on equine neutrophils.
- The researchers assessed this antigen expression using immunofluorescence flow cytometry, a technique which enables them to measure physical and chemical characteristics of cells or particles.
- Evaluations were performed on how interactions with various substances such as zymosan-activated serum (ZAS), phorbol 12-myristate 13-acetate (PMA), bacterial lipopolysaccharide (LPS) and different anti-inflammatory drugs, can modulate this antigen expression.
Key Findings
- Stimulation using ZAS or PMA was found to enhance the expression of the antigen which MoAb 60.3 recognizes.
- Inhibition of ZAS-stimulated aggregation was observed upon pre-treatment of equine neutrophils with MoAb 60.3, suggesting this antibody recognizes a functional feature of the cells related to adhesion.
- In experiments with bacterial LPS, a moderate increase in MoAb 60.3 binding was noticed, reflecting a possible enhancement in the expression of the antigen on the neutrophils. Pre-incubation with LPS also led to a slight increase in MoAb 60.3 binding after stimulation with ZAS.
- Pre-treatment of neutrophils with anti-inflammatory drugs obstructed some extent of ZAS-stimulated neutrophil adhesion onto layers of equine endothelium cells. But, it was observed that the effect of such drugs did not correlate well with the overall expression levels of the antigen recognized by MoAb 60.3.
Conclusion
- The results suggest that horse neutrophils express a functional adhesion molecule similar to those present on human neutrophils and that LPS might enhance the expression of this surface antigen.
- However, the expression of this adhesion-related surface antigen doesn’t correspond well with levels of drug-induced diminished adhesion of neutrophils to endothelium, suggesting more complex mechanisms may be at play in the immune response modulation by the drugs.
Cite This Article
APA
Bochsler PN, Slauson DO, Neilsen NR.
(1990).
Modulation of an adhesion-related surface antigen on equine neutrophils by bacterial lipopolysaccharide and antiinflammatory drugs.
J Leukoc Biol, 48(4), 306-315.
https://doi.org/10.1002/jlb.48.4.306 Publication
Researcher Affiliations
- Department of Pathology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853.
MeSH Terms
- Animals
- Anti-Inflammatory Agents / pharmacology
- Antibodies, Monoclonal
- Cell Adhesion
- Cells, Cultured
- Dimethyl Sulfoxide
- Endothelium / metabolism
- Flow Cytometry
- Horses
- Lipopolysaccharides / pharmacology
- Neutrophils / immunology
- Receptors, Leukocyte-Adhesion / immunology
- Tetradecanoylphorbol Acetate
- Up-Regulation
- Zymosan / pharmacology
Citations
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