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Related Experiment Videos

Defensin gene expression in the cornea

J D Gottsch1, Q Li, M F Ashraf

  • 1Wilmer Ophthalmological Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA.

Current Eye Research
|December 10, 1998
PubMed
Summary

Defensins, antimicrobial peptides, are present in diseased human corneas and produced by corneal cells when stimulated by inflammatory cytokines. This suggests a role in fighting microbial keratitis.

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Area of Science:

  • Ophthalmology
  • Immunology
  • Molecular Biology

Background:

  • Defensins are antimicrobial peptides crucial for innate immunity.
  • Their role in corneal defense, particularly in disease states, is not fully understood.

Purpose of the Study:

  • To investigate the expression of defensin genes in human corneas.
  • To determine if bovine corneal keratocytes can produce defensins.

Main Methods:

  • In situ hybridization and immunohistochemistry for human corneal tissue.
  • Cytokine stimulation (IL-1alpha, TNFalpha) of cultured bovine keratocytes.
  • Reverse transcription polymerase chain reaction (RT-PCR) and Southern blotting for gene expression analysis.

Main Results:

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  • Defensin mRNA and protein were absent in normal human corneal stroma but detected in diseased corneas (rejected transplants, postinfectious keratitis).
  • Interleukin-1alpha (IL-1alpha) potently induced defensin gene expression in keratocytes; Tumor Necrosis Factor-alpha (TNFalpha) showed a weaker induction.
  • RT-PCR confirmed specific defensin gene amplification in cytokine-stimulated keratocytes.

Conclusions:

  • Human corneas express defensins, particularly during inflammatory conditions.
  • Corneal keratocytes possess the capability to produce defensin mRNA in response to inflammatory cytokines.
  • Defensin release by keratocytes may play a significant role in the host defense against microbial keratitis.