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Evaluation of limbal stem cell deficiency: a new diagnostic tool based on keratin 13 immunostaining in corneal impression cytology
Author(s) -
POLI M,
JUSTIN V,
DAMOUR O,
BURILLON C
Publication year - 2012
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2012.t041.x
Subject(s) - medicine , immunostaining , cytology , pathology , ophthalmology , corneal diseases , cornea , immunohistochemistry
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. Conclusion 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.