Expression and release of tumor necrosis factor-alpha by explants of mouse cornea

Maki Sekine-Okano, Rudolf Lucas, Duri Rungger, Toon De Kesel, Georges E. Grau, Peter M. Leuenberger, Elisabeth Rungger-Brändle

Research output: Contribution to journalArticle

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Abstract

Purpose. To elucidate a possible target of immunosuppressive agents widely used in the treatment of corneal disorders, the authors determined whether corneal cells are capable of expressing and releasing tumor necrosis factor- alpha (TNFα) on lipopolysaccharide (LPS) stimulation, and they investigated whether TNFα production can be modulated by pharmacologic agents. Methods. Trephined central corneas from C57BL/6 mice were kept in culture for 3 days. Release of TNFα after a 24-hour stimulation with LPS (1 μg/ml) into the culture medium was determined both by bioassay and by enzyme-linked immunosorbent assay. Expression of TNFα mRNA after 6-hour stimulation was examined by polymerase chain reaction. Immunofluorescent staining on cryostat sections of cultured corneas was performed to localize TNFα in the tissue. Corneal explants were retreated with immunosuppressive agents (prednisolone, budesonide, cyclosporin A) for 48 hours, followed by 6- or 24-hour stimulation with LPS in the continuous presence of the agents. Results. Lipopolysaccharide stimulated TNF∅ release into the culture medium. The addition of budesonide (10-7M) or prednisolone (10-6 M) significantly inhibited LPS-induced TNFσ release, whereas cyclosporin A (10-7-10-5 M) had no marked effect. levels of TNFα mRNA in corneal explants increased fivefold after stimulation with LPS. Immunohistochemical staining revealed that TNFα was expressed in the epithelia cells. Budesonide markedly decreased mRNA expression and abolished immunostaining of TNFα stimulated by LPS. Conclusions. TNFα is produced and release by the epithelial cells of mouse central cornea in response to LPS. Contrary to cyclosporin A corticosteroids such as prednisolone and budesonide potently inhibit TNFα production.

Original languageEnglish (US)
Pages (from-to)1302-1310
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number7
StatePublished - Jun 1 1996

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Cornea
Tumor Necrosis Factor-alpha
Lipopolysaccharides
Budesonide
Prednisolone
Cyclosporine
Immunosuppressive Agents
Messenger RNA
Culture Media
Staining and Labeling
Inbred C57BL Mouse
Biological Assay
Adrenal Cortex Hormones
Epithelium
Epithelial Cells
Enzyme-Linked Immunosorbent Assay
Polymerase Chain Reaction

Keywords

  • corneal epithelium
  • corticosteroids
  • cyclosporin
  • cytokine
  • tumor necrosis factor-alpha (TNFα)

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Sekine-Okano, M., Lucas, R., Rungger, D., De Kesel, T., Grau, G. E., Leuenberger, P. M., & Rungger-Brändle, E. (1996). Expression and release of tumor necrosis factor-alpha by explants of mouse cornea. Investigative Ophthalmology and Visual Science, 37(7), 1302-1310.

Expression and release of tumor necrosis factor-alpha by explants of mouse cornea. / Sekine-Okano, Maki; Lucas, Rudolf; Rungger, Duri; De Kesel, Toon; Grau, Georges E.; Leuenberger, Peter M.; Rungger-Brändle, Elisabeth.

In: Investigative Ophthalmology and Visual Science, Vol. 37, No. 7, 01.06.1996, p. 1302-1310.

Research output: Contribution to journalArticle

Sekine-Okano, M, Lucas, R, Rungger, D, De Kesel, T, Grau, GE, Leuenberger, PM & Rungger-Brändle, E 1996, 'Expression and release of tumor necrosis factor-alpha by explants of mouse cornea', Investigative Ophthalmology and Visual Science, vol. 37, no. 7, pp. 1302-1310.
Sekine-Okano M, Lucas R, Rungger D, De Kesel T, Grau GE, Leuenberger PM et al. Expression and release of tumor necrosis factor-alpha by explants of mouse cornea. Investigative Ophthalmology and Visual Science. 1996 Jun 1;37(7):1302-1310.
Sekine-Okano, Maki ; Lucas, Rudolf ; Rungger, Duri ; De Kesel, Toon ; Grau, Georges E. ; Leuenberger, Peter M. ; Rungger-Brändle, Elisabeth. / Expression and release of tumor necrosis factor-alpha by explants of mouse cornea. In: Investigative Ophthalmology and Visual Science. 1996 ; Vol. 37, No. 7. pp. 1302-1310.
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abstract = "Purpose. To elucidate a possible target of immunosuppressive agents widely used in the treatment of corneal disorders, the authors determined whether corneal cells are capable of expressing and releasing tumor necrosis factor- alpha (TNFα) on lipopolysaccharide (LPS) stimulation, and they investigated whether TNFα production can be modulated by pharmacologic agents. Methods. Trephined central corneas from C57BL/6 mice were kept in culture for 3 days. Release of TNFα after a 24-hour stimulation with LPS (1 μg/ml) into the culture medium was determined both by bioassay and by enzyme-linked immunosorbent assay. Expression of TNFα mRNA after 6-hour stimulation was examined by polymerase chain reaction. Immunofluorescent staining on cryostat sections of cultured corneas was performed to localize TNFα in the tissue. Corneal explants were retreated with immunosuppressive agents (prednisolone, budesonide, cyclosporin A) for 48 hours, followed by 6- or 24-hour stimulation with LPS in the continuous presence of the agents. Results. Lipopolysaccharide stimulated TNF∅ release into the culture medium. The addition of budesonide (10-7M) or prednisolone (10-6 M) significantly inhibited LPS-induced TNFσ release, whereas cyclosporin A (10-7-10-5 M) had no marked effect. levels of TNFα mRNA in corneal explants increased fivefold after stimulation with LPS. Immunohistochemical staining revealed that TNFα was expressed in the epithelia cells. Budesonide markedly decreased mRNA expression and abolished immunostaining of TNFα stimulated by LPS. Conclusions. TNFα is produced and release by the epithelial cells of mouse central cornea in response to LPS. Contrary to cyclosporin A corticosteroids such as prednisolone and budesonide potently inhibit TNFα production.",
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AU - Leuenberger, Peter M.

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AB - Purpose. To elucidate a possible target of immunosuppressive agents widely used in the treatment of corneal disorders, the authors determined whether corneal cells are capable of expressing and releasing tumor necrosis factor- alpha (TNFα) on lipopolysaccharide (LPS) stimulation, and they investigated whether TNFα production can be modulated by pharmacologic agents. Methods. Trephined central corneas from C57BL/6 mice were kept in culture for 3 days. Release of TNFα after a 24-hour stimulation with LPS (1 μg/ml) into the culture medium was determined both by bioassay and by enzyme-linked immunosorbent assay. Expression of TNFα mRNA after 6-hour stimulation was examined by polymerase chain reaction. Immunofluorescent staining on cryostat sections of cultured corneas was performed to localize TNFα in the tissue. Corneal explants were retreated with immunosuppressive agents (prednisolone, budesonide, cyclosporin A) for 48 hours, followed by 6- or 24-hour stimulation with LPS in the continuous presence of the agents. Results. Lipopolysaccharide stimulated TNF∅ release into the culture medium. The addition of budesonide (10-7M) or prednisolone (10-6 M) significantly inhibited LPS-induced TNFσ release, whereas cyclosporin A (10-7-10-5 M) had no marked effect. levels of TNFα mRNA in corneal explants increased fivefold after stimulation with LPS. Immunohistochemical staining revealed that TNFα was expressed in the epithelia cells. Budesonide markedly decreased mRNA expression and abolished immunostaining of TNFα stimulated by LPS. Conclusions. TNFα is produced and release by the epithelial cells of mouse central cornea in response to LPS. Contrary to cyclosporin A corticosteroids such as prednisolone and budesonide potently inhibit TNFα production.

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