Endotoxin-induced leukocyte accumulation in aqueous fluid of rats is decreased by a small molecule selectin antagonist

Ronald Tilton, S. J. Sherwood, R. J. Bjercke, P. J. Beck, B. Dupré, E. T H Yeh, T. P. Kogan

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Purpose. Selectins are a family of carbohydrate binding proteins expressed on surfaces of endothelium (E- and P-selectin), platelets (P-selectin) and leukocytes (L-selectin). Since binding of the sialyl LewisX oligosaccharides on leukocytes to selectins initiates cellular recruitment in response to inflammatory stimuli, these experiments investigated a possible role for selectins in leukocyte accumulation in aqueous fluid following exposure to endotoxin. Methods. A non-oligosaccharide selectin inhibitor, TBC265, was assayed for its ability to inhibit binding of HL60 cells (which express surface sialyl LewisX-bearing glycoproteins) to recombinant E-, P-, and L-selectin-IgG fusion proteins bound to magnetic beads. To induce uveitis, 200 μg LPS (Salmonella minnesota) was injected as a divided dose into both hind footpads of male, 175-200 g Lewis rats anesthetized with halothane. Vehicle (0.5 ml PBS) or TBC265 (30 mg/0.5 ml PBS) was injected s.c. 3, 5, and 7 hours post LPS injection, and rats were killed after 24 h. Total recoverable neutrophils (PMN) were calculated from a total cell count and differential cell determination using aqueous fluid pooled from both eyes. Results. IC50 values for TBC265 inhibition of HL60 cell binding to E-, P-, and L-selectin-IgG fusion proteins was 3, 2, and 2 mM, respectively. Endotoxin increased recoverable PMN from essentially 0 in controls to 9,400 ± 1,940 (mean ± SEM; n=26); TBC265 decreased recoverable PMN 80 % to 1790 ± 516 (n=14; p<0.001). Conclusions. Results from these experiments suggest that selectin adhesion molecules play a role in the response of the anterior uvea to endotoxin, and indicate that small molecule mimetics of sialyl LewisX oligosaccharides may have therapeutic potential in preventing sequelae of acute inflammation.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number3
StatePublished - Feb 15 1996
Externally publishedYes

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Selectins
P-Selectin
Endotoxins
Leukocytes
E-Selectin
Blood Platelets
HL-60 Cells
Oligosaccharides
Immunoglobulin G
Uvea
Uveitis
Halothane
Salmonella
Inhibitory Concentration 50
Endothelium
Glycoproteins
Proteins
Neutrophils
Cell Count
Inflammation

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Tilton, R., Sherwood, S. J., Bjercke, R. J., Beck, P. J., Dupré, B., Yeh, E. T. H., & Kogan, T. P. (1996). Endotoxin-induced leukocyte accumulation in aqueous fluid of rats is decreased by a small molecule selectin antagonist. Investigative Ophthalmology and Visual Science, 37(3).

Endotoxin-induced leukocyte accumulation in aqueous fluid of rats is decreased by a small molecule selectin antagonist. / Tilton, Ronald; Sherwood, S. J.; Bjercke, R. J.; Beck, P. J.; Dupré, B.; Yeh, E. T H; Kogan, T. P.

In: Investigative Ophthalmology and Visual Science, Vol. 37, No. 3, 15.02.1996.

Research output: Contribution to journalArticle

Tilton, Ronald ; Sherwood, S. J. ; Bjercke, R. J. ; Beck, P. J. ; Dupré, B. ; Yeh, E. T H ; Kogan, T. P. / Endotoxin-induced leukocyte accumulation in aqueous fluid of rats is decreased by a small molecule selectin antagonist. In: Investigative Ophthalmology and Visual Science. 1996 ; Vol. 37, No. 3.
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abstract = "Purpose. Selectins are a family of carbohydrate binding proteins expressed on surfaces of endothelium (E- and P-selectin), platelets (P-selectin) and leukocytes (L-selectin). Since binding of the sialyl LewisX oligosaccharides on leukocytes to selectins initiates cellular recruitment in response to inflammatory stimuli, these experiments investigated a possible role for selectins in leukocyte accumulation in aqueous fluid following exposure to endotoxin. Methods. A non-oligosaccharide selectin inhibitor, TBC265, was assayed for its ability to inhibit binding of HL60 cells (which express surface sialyl LewisX-bearing glycoproteins) to recombinant E-, P-, and L-selectin-IgG fusion proteins bound to magnetic beads. To induce uveitis, 200 μg LPS (Salmonella minnesota) was injected as a divided dose into both hind footpads of male, 175-200 g Lewis rats anesthetized with halothane. Vehicle (0.5 ml PBS) or TBC265 (30 mg/0.5 ml PBS) was injected s.c. 3, 5, and 7 hours post LPS injection, and rats were killed after 24 h. Total recoverable neutrophils (PMN) were calculated from a total cell count and differential cell determination using aqueous fluid pooled from both eyes. Results. IC50 values for TBC265 inhibition of HL60 cell binding to E-, P-, and L-selectin-IgG fusion proteins was 3, 2, and 2 mM, respectively. Endotoxin increased recoverable PMN from essentially 0 in controls to 9,400 ± 1,940 (mean ± SEM; n=26); TBC265 decreased recoverable PMN 80 {\%} to 1790 ± 516 (n=14; p<0.001). Conclusions. Results from these experiments suggest that selectin adhesion molecules play a role in the response of the anterior uvea to endotoxin, and indicate that small molecule mimetics of sialyl LewisX oligosaccharides may have therapeutic potential in preventing sequelae of acute inflammation.",
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AU - Tilton, Ronald

AU - Sherwood, S. J.

AU - Bjercke, R. J.

AU - Beck, P. J.

AU - Dupré, B.

AU - Yeh, E. T H

AU - Kogan, T. P.

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