Suppression of nuclear factor kappa B and CD18-mediated leukocyte adhesion to the corneal endothelium by dexamethasone

Masaru Shimoyama, Shigeto Shimmura, Kazuo Tsubota, Yoshihisa Oguchi

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Purpose. To demonstrate that leukocyte adhesion to cultured corneal endothelial cells is mediated by the CD18 antigen, and to determine whether dexamethasone directly suppresses adhesion by inhibiting activation of nuclear factor kappa B (NFκB). Methods. Cultured bovine corneal endothelium was stimulated for 6 hours by 40 u/ml tumor necrosis factor alpha (TNFα). Dexamethasone was added 1 hour before TNFα stimulation in the dexamethasone group. After stimulation, neutrophils separated from a healthy human volunteer were added with or without anti-CD18 antibody. The culture plate was settled for 15 minutes at 37°C, and then neutrophils were activated by N-formylmethionyl-leucyl-phenylalanine for 5 minutes. Nonadherent neutrophils were removed by sealing and inverting the culture well. The intracellular localization of NFkB after TNFα simulation was determined by confocal immunocytochemistry using an anti-p65 antibody. Results. Neutrophil adhesion to cultured corneal endothelial cells increased significantly on exposure to TNFa (451.4 ±45.4 cells/mm2, n = 16) compared to control (156.7 ± 27.3 cells/mm2 n = 16, P <0.01). This increased adhesion was suppressed by the addition of anti-CD18 antibody (157.6 ± 25.1 cells/mm2, n = 8, P < 0.01) and by pretreatment with 10-7 M dexamethaSone (207.9 ± 31.5 cells/mm2, n = 10, P < 0.01). Immunocytochemistry 60 minutes after stimulation revealed that NFκB was located in the cytoplasm in unstimulated cells; however, the addition of TNFα caused NFκB to translocate into the nucleus. Pretreatment with dexamethasone tapered NFκB translocation into the nucleus. Conclusions. Leukocyte adhesion to the corneal endothelium was shown to be mediated by CD18 expressed on activated leukocytes. Pretreatment of the endothelium with dexamethasone inhibited leukocyte adhesion; this may be due in part to the suppression of NFκB entry into the nucleus.

Original languageEnglish
Pages (from-to)2427-2431
Number of pages5
JournalInvestigative Ophthalmology and Visual Science
Volume38
Issue number11
Publication statusPublished - 1997 Oct

Fingerprint

Corneal Endothelium
NF-kappa B
Dexamethasone
Leukocytes
Neutrophils
Tumor Necrosis Factor-alpha
Anti-Idiotypic Antibodies
CD18 Antigens
Endothelial Cells
Immunohistochemistry
N-Formylmethionine Leucyl-Phenylalanine
Endothelium
Healthy Volunteers
Cytoplasm

Keywords

  • CD18
  • Corneal endothelium
  • Nuclear factor kappa B
  • Polymorphonucleocyte
  • Tumor necrosis factor alpha

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Suppression of nuclear factor kappa B and CD18-mediated leukocyte adhesion to the corneal endothelium by dexamethasone. / Shimoyama, Masaru; Shimmura, Shigeto; Tsubota, Kazuo; Oguchi, Yoshihisa.

In: Investigative Ophthalmology and Visual Science, Vol. 38, No. 11, 10.1997, p. 2427-2431.

Research output: Contribution to journalArticle

@article{b44ff39b35034e19a3b5b7ce67dbcf12,
title = "Suppression of nuclear factor kappa B and CD18-mediated leukocyte adhesion to the corneal endothelium by dexamethasone",
abstract = "Purpose. To demonstrate that leukocyte adhesion to cultured corneal endothelial cells is mediated by the CD18 antigen, and to determine whether dexamethasone directly suppresses adhesion by inhibiting activation of nuclear factor kappa B (NFκB). Methods. Cultured bovine corneal endothelium was stimulated for 6 hours by 40 u/ml tumor necrosis factor alpha (TNFα). Dexamethasone was added 1 hour before TNFα stimulation in the dexamethasone group. After stimulation, neutrophils separated from a healthy human volunteer were added with or without anti-CD18 antibody. The culture plate was settled for 15 minutes at 37°C, and then neutrophils were activated by N-formylmethionyl-leucyl-phenylalanine for 5 minutes. Nonadherent neutrophils were removed by sealing and inverting the culture well. The intracellular localization of NFkB after TNFα simulation was determined by confocal immunocytochemistry using an anti-p65 antibody. Results. Neutrophil adhesion to cultured corneal endothelial cells increased significantly on exposure to TNFa (451.4 ±45.4 cells/mm2, n = 16) compared to control (156.7 ± 27.3 cells/mm2 n = 16, P <0.01). This increased adhesion was suppressed by the addition of anti-CD18 antibody (157.6 ± 25.1 cells/mm2, n = 8, P < 0.01) and by pretreatment with 10-7 M dexamethaSone (207.9 ± 31.5 cells/mm2, n = 10, P < 0.01). Immunocytochemistry 60 minutes after stimulation revealed that NFκB was located in the cytoplasm in unstimulated cells; however, the addition of TNFα caused NFκB to translocate into the nucleus. Pretreatment with dexamethasone tapered NFκB translocation into the nucleus. Conclusions. Leukocyte adhesion to the corneal endothelium was shown to be mediated by CD18 expressed on activated leukocytes. Pretreatment of the endothelium with dexamethasone inhibited leukocyte adhesion; this may be due in part to the suppression of NFκB entry into the nucleus.",
keywords = "CD18, Corneal endothelium, Nuclear factor kappa B, Polymorphonucleocyte, Tumor necrosis factor alpha",
author = "Masaru Shimoyama and Shigeto Shimmura and Kazuo Tsubota and Yoshihisa Oguchi",
year = "1997",
month = "10",
language = "English",
volume = "38",
pages = "2427--2431",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "11",

}

TY - JOUR

T1 - Suppression of nuclear factor kappa B and CD18-mediated leukocyte adhesion to the corneal endothelium by dexamethasone

AU - Shimoyama, Masaru

AU - Shimmura, Shigeto

AU - Tsubota, Kazuo

AU - Oguchi, Yoshihisa

PY - 1997/10

Y1 - 1997/10

N2 - Purpose. To demonstrate that leukocyte adhesion to cultured corneal endothelial cells is mediated by the CD18 antigen, and to determine whether dexamethasone directly suppresses adhesion by inhibiting activation of nuclear factor kappa B (NFκB). Methods. Cultured bovine corneal endothelium was stimulated for 6 hours by 40 u/ml tumor necrosis factor alpha (TNFα). Dexamethasone was added 1 hour before TNFα stimulation in the dexamethasone group. After stimulation, neutrophils separated from a healthy human volunteer were added with or without anti-CD18 antibody. The culture plate was settled for 15 minutes at 37°C, and then neutrophils were activated by N-formylmethionyl-leucyl-phenylalanine for 5 minutes. Nonadherent neutrophils were removed by sealing and inverting the culture well. The intracellular localization of NFkB after TNFα simulation was determined by confocal immunocytochemistry using an anti-p65 antibody. Results. Neutrophil adhesion to cultured corneal endothelial cells increased significantly on exposure to TNFa (451.4 ±45.4 cells/mm2, n = 16) compared to control (156.7 ± 27.3 cells/mm2 n = 16, P <0.01). This increased adhesion was suppressed by the addition of anti-CD18 antibody (157.6 ± 25.1 cells/mm2, n = 8, P < 0.01) and by pretreatment with 10-7 M dexamethaSone (207.9 ± 31.5 cells/mm2, n = 10, P < 0.01). Immunocytochemistry 60 minutes after stimulation revealed that NFκB was located in the cytoplasm in unstimulated cells; however, the addition of TNFα caused NFκB to translocate into the nucleus. Pretreatment with dexamethasone tapered NFκB translocation into the nucleus. Conclusions. Leukocyte adhesion to the corneal endothelium was shown to be mediated by CD18 expressed on activated leukocytes. Pretreatment of the endothelium with dexamethasone inhibited leukocyte adhesion; this may be due in part to the suppression of NFκB entry into the nucleus.

AB - Purpose. To demonstrate that leukocyte adhesion to cultured corneal endothelial cells is mediated by the CD18 antigen, and to determine whether dexamethasone directly suppresses adhesion by inhibiting activation of nuclear factor kappa B (NFκB). Methods. Cultured bovine corneal endothelium was stimulated for 6 hours by 40 u/ml tumor necrosis factor alpha (TNFα). Dexamethasone was added 1 hour before TNFα stimulation in the dexamethasone group. After stimulation, neutrophils separated from a healthy human volunteer were added with or without anti-CD18 antibody. The culture plate was settled for 15 minutes at 37°C, and then neutrophils were activated by N-formylmethionyl-leucyl-phenylalanine for 5 minutes. Nonadherent neutrophils were removed by sealing and inverting the culture well. The intracellular localization of NFkB after TNFα simulation was determined by confocal immunocytochemistry using an anti-p65 antibody. Results. Neutrophil adhesion to cultured corneal endothelial cells increased significantly on exposure to TNFa (451.4 ±45.4 cells/mm2, n = 16) compared to control (156.7 ± 27.3 cells/mm2 n = 16, P <0.01). This increased adhesion was suppressed by the addition of anti-CD18 antibody (157.6 ± 25.1 cells/mm2, n = 8, P < 0.01) and by pretreatment with 10-7 M dexamethaSone (207.9 ± 31.5 cells/mm2, n = 10, P < 0.01). Immunocytochemistry 60 minutes after stimulation revealed that NFκB was located in the cytoplasm in unstimulated cells; however, the addition of TNFα caused NFκB to translocate into the nucleus. Pretreatment with dexamethasone tapered NFκB translocation into the nucleus. Conclusions. Leukocyte adhesion to the corneal endothelium was shown to be mediated by CD18 expressed on activated leukocytes. Pretreatment of the endothelium with dexamethasone inhibited leukocyte adhesion; this may be due in part to the suppression of NFκB entry into the nucleus.

KW - CD18

KW - Corneal endothelium

KW - Nuclear factor kappa B

KW - Polymorphonucleocyte

KW - Tumor necrosis factor alpha

UR - http://www.scopus.com/inward/record.url?scp=0030821336&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030821336&partnerID=8YFLogxK

M3 - Article

VL - 38

SP - 2427

EP - 2431

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 11

ER -