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Investigation of β defensin gene expression in the ocular anterior segment by semiquantitative RT-PCR
  1. Ordan J Lehmanna,
  2. Imran R Hussainb,
  3. Peter J Wattc
  1. aMoorfields Eye Hospital, London, bUniversity Medicine, University of Southampton, cDepartment of Molecular Microbiology, University of Southampton
  1. Mr O J Lehmann, Glaucoma Research Unit, Moorfields Eye Hospital, City Road, London EC1V 2PD ojlehmann{at}yahoo.com

Abstract

AIM To determine if β 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 β defensin mRNA by semiquantitative RT-PCR and the mRNA standardised for cell numbers.

RESULTS RT-PCR amplified β defensin 1 mRNA from all lens capsule (six) and corneal (five) samples and all but one of the conjunctival (six) and iris samples (six). β 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 β defensin 2 not expressed intraocularly and higher levels of β defensin 1 than β defensin 2 expressed in extraocular tissue. The implication is that β defensin 1 is constitutively produced in ocular tissues and represents a key component of the innate immune system.

  • defensins
  • gene expression
  • polymerase chain reaction

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