March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Evaluation Of Limbal Stem Cell Deficiency: A New Diagnostic Tool Based On Keratin 13 Immunostaining In Corneal Impression Cytology
Author Affiliations & Notes
  • Muriel Poli
    Ophthalmology Dept,
    CHU Lyon, Lyon, France
  • Virginie Justin
    Laboratoire des Greffes et Substituts Cutanés, Pavillon I,
    CHU Lyon, Lyon, France
  • Odile Damour
    Laboratoire des Greffes et Substituts Cutanés, Pavillon I,
    CHU Lyon, Lyon, France
  • Carole Burillon
    Ophthalmology Dept,
    CHU Lyon, Lyon, France
  • Footnotes
    Commercial Relationships  Muriel Poli, None; Virginie Justin, None; Odile Damour, None; Carole Burillon, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 3515. doi:
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      Muriel Poli, Virginie Justin, Odile Damour, Carole Burillon; Evaluation Of Limbal Stem Cell Deficiency: A New Diagnostic Tool Based On Keratin 13 Immunostaining In Corneal Impression Cytology. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3515.

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Abstract

Purpose: : The aim of this study was to develop a validated, reliable and minimally invasive technique for diagnosing limbal stem cell deficiency (LSCD) using immunocytochemical detection of conjunctival and corneal keratins on epithelial cells collected by impression cytology (IC).

Methods: : After validation of the labelling techniques on a cohort of ten healthy control patients, keratins K12, K13 and K19 were labelled on corneal IC of ten eyes suspected of LSCD. Positive scores for the conjunctival markers K13/K19, combined with the rarity of the corneal marker K12, were diagnostic proof of LSCD

Results: : IC is a reliable and noninvasive technique for collecting epithelial cells. The labelling validation phase eliminated K3 labelling due to lack of corneal specificity.Among the patients with suspected LSCD, nine samples were diagnosed with LSCD, which was severe (K12 negative) in eight cases and mild (K12 positive) in one case. One sample could not be analysed due to insufficient cells.

Conclusions: : The immunocytochemical search for the K13/K19 pair using corneal IC provides a simple and reliable method for diagnosing LSCD, while the level of K12 provides a score of disease severity. On the other hand, we question the corneal specificity of K3 as conventionally established. Finally, to our knowledge, this is the first study detecting K13 on a conjunctival IC specimen as a marker of conjunctival differentiation.

Keywords: cornea: epithelium • clinical laboratory testing 
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