Investigation of beta defensin gene expression in the ocular anterior segment by semiquantitative RT-PCR

Br J Ophthalmol. 2000 May;84(5):523-6. doi: 10.1136/bjo.84.5.523.

Abstract

Aim: To determine if beta defensins are expressed in the anterior segment of the eye and to determine the temporal pattern of expression using a real time semiquantitative reverse transcription polymerase chain reaction (RT-PCR).

Methods: Ocular tissue (corneal epithelium, conjunctiva, iris, and lens capsule) was collected from 23 patients undergoing surgery. Serial corneal or conjunctival impression cytology was performed on a separate group of 10 patients undergoing corneal tunnel phacoemulsification or trabeculectomy. The samples were analysed for beta defensin mRNA by semiquantitative RT-PCR and the mRNA standardised for cell numbers.

Results: RT-PCR amplified beta defensin 1 mRNA from all lens capsule (six) and corneal (five) samples and all but one of the conjunctival (six) and iris samples (six). beta Defensin 2 mRNA was amplified from three of five corneal, two of six conjunctival, and none of the iris or capsule samples. The impression cytology samples demonstrated a decline in defensin expression over the three time points studied. There were no false positive results from either the no-RT or negative control samples.

Conclusions: This preliminary study confirms that natural antibacterial peptides are expressed in the anterior segment of the eye. There appears to be a pattern to the expression with inducible beta defensin 2 not expressed intraocularly and higher levels of beta defensin 1 than beta defensin 2 expressed in extraocular tissue. The implication is that beta defensin 1 is constitutively produced in ocular tissues and represents a key component of the innate immune system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anterior Eye Segment / metabolism*
  • Conjunctiva / metabolism
  • Defensins
  • Epithelium, Corneal / metabolism
  • Gene Expression
  • Humans
  • Iris / metabolism
  • Lens Capsule, Crystalline / metabolism
  • Proteins / genetics
  • Proteins / metabolism*
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Defensins
  • Proteins
  • RNA, Messenger