Defensin gene expression in the cornea

John D Gottsch, Q. Li, M. F. Ashraf, T. P. O'Brien, W. J. Stark, S. H. Liu

Research output: Contribution to journalArticle

Abstract

Purpose. To determine whether defensin genes are expressed in human corneas and bovine corneal keratocytes. Methods. In situ hybridization and immunohistochemistry were used to localize defensin mRNA and protein in normal and diseased human corneas. Cultured bovine keratocytes were stimulated with IL-1α or TNFα to determine whether defensin mRNA production occurred. Reverse transcription polymerase chain reaction (RT-PCR) was performed to amplify defensin cDNA from cytokine-induced keratocytes, and Southern blots were used to verify the specificity of RT-PCR amplification products. Results. Defensin mRNA and protein were not detected in normal human corneal stroma, but were readily detectable in the corneal stroma in cases of rejected transplants and postinfectious keratitis. IL-1α was a potent inducer of defensin gene expression in keratocytes, which began 12 h after challenge and peaked at 18 to 24 h. TNFα weakly induced defensin mRNA in keratocytes at about 18 h. Southern blots of the RT-CR products probed with an oligonucleotide complementary to internal sequences of defensin demonstrated the appropriately sized products (198 bp) specific for defensin. Conclusions. This report demonstrates the presence of defensin in the human cornea and the capacity of corneal keratocytes to produce defensin mRNA in response to IL-1α and TNFα. Release of defensins by keratocytes in response to cytokines elaborated in corneal inflammation may contribute to the host defense response in microbial keratitis.

Original languageEnglish (US)
Pages (from-to)1082-1086
Number of pages5
JournalCurrent Eye Research
Volume17
Issue number11
DOIs
StatePublished - 1998

Fingerprint

Defensins
Cornea
Gene Expression
Corneal Keratocytes
Messenger RNA
Interleukin-1
Corneal Stroma
Keratitis
Southern Blotting
Reverse Transcription
Cytokines
Polymerase Chain Reaction
Oligonucleotides
In Situ Hybridization
Proteins

Keywords

  • Cornea
  • Cytokines
  • Defensin
  • Helminthiasis
  • Neutrophils

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

Gottsch, J. D., Li, Q., Ashraf, M. F., O'Brien, T. P., Stark, W. J., & Liu, S. H. (1998). Defensin gene expression in the cornea. Current Eye Research, 17(11), 1082-1086. https://doi.org/10.1076/ceyr.17.11.1082.5235

Defensin gene expression in the cornea. / Gottsch, John D; Li, Q.; Ashraf, M. F.; O'Brien, T. P.; Stark, W. J.; Liu, S. H.

In: Current Eye Research, Vol. 17, No. 11, 1998, p. 1082-1086.

Research output: Contribution to journalArticle

Gottsch, JD, Li, Q, Ashraf, MF, O'Brien, TP, Stark, WJ & Liu, SH 1998, 'Defensin gene expression in the cornea', Current Eye Research, vol. 17, no. 11, pp. 1082-1086. https://doi.org/10.1076/ceyr.17.11.1082.5235
Gottsch JD, Li Q, Ashraf MF, O'Brien TP, Stark WJ, Liu SH. Defensin gene expression in the cornea. Current Eye Research. 1998;17(11):1082-1086. https://doi.org/10.1076/ceyr.17.11.1082.5235
Gottsch, John D ; Li, Q. ; Ashraf, M. F. ; O'Brien, T. P. ; Stark, W. J. ; Liu, S. H. / Defensin gene expression in the cornea. In: Current Eye Research. 1998 ; Vol. 17, No. 11. pp. 1082-1086.
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