title: Immunocytochemical Diagnosis of Limbal Stem Cell Deficiency: Comparative Analysis of Current Corneal and Conjunctival Biomarkers.
abstract: OBJECTIVE
  To evaluate and compare corneal and conjunctival biomarkers for immunocytochemical diagnosis of limbal stem cell deficiency (LSCD).
  
  
  METHODS
  In accordance with the current literature, we selected K12 as the corneal biomarker and K7/K13/K19/MUC5AC as the conjunctival ones. The specificity and accuracy for each biomarker were assessed and compared on 10 healthy subjects and tissues of deceased donors. Twelve eyes of 9 patients clinically suspected of LSCD were enrolled. Epithelial cells (ECs) from the central cornea were collected using impression cytology (IC) and assessed for each biomarker. The presence of conjunctival cells in the central cornea was diagnostic proof of LSCD, whereas the detection of corneal residual cells would quantify the degree of LSCD.
  
  
  RESULTS
  K12 and K7/K13/MUC5AC are, respectively, highly specific of corneal and conjunctival differentiation, whereas K19 is not. Normal corneal ECs are not desquamative enough to be suitable for IC. Among 12 eyes with suspected LSCD, 84% (10 of 12) of IC samples were suitable for analysis. K3/K7/K19 immunostaining was positive in 100%, MUC5AC in 40%, and K12 was never observed.
  
  
  CONCLUSIONS
  Clinical examination can lead to misdiagnosis of LSCD. Immunocytochemical detection of K7/K13 on corneal ECs collected by IC is reproducible, noninvasive, and highly effective in this indication, but without any quantification of the degree of the disease. This time-consuming technique requires skilled technicians and laboratory facilities, reserving it for planned limbal reconstruction.
authors.affiliation: *Clinique Ophtalmologique, Pavillon C, Hôpital Edouard Herriot, Lyon, France; and †Laboratoire des Greffes et Substituts Cutanés, Pavillon I, Hôpital Edouard Herriot, Lyon, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Poli
authors.forename: Carole
authors.initials: C
authors.lastname: Burillon
authors.forename: Celine
authors.initials: C
authors.lastname: Auxenfans
authors.forename: Marie-Rose
authors.initials: MR
authors.lastname: Rovere
authors.forename: Odile
authors.initials: O
authors.lastname: Damour
mesh_terms: D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000378:metabolism*; D003228:Conjunctiva / Q000378:metabolism*; D003316:Corneal Diseases / Q000175:diagnosis* / Q000378:metabolism; D019573:Epithelium, Corneal / Q000378:metabolism*; D005136:Eye Proteins / Q000378:metabolism*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D053551:Keratin-12 / Q000378:metabolism; D053535:Keratin-13 / Q000378:metabolism; D053539:Keratin-19 / Q000378:metabolism; D053552:Keratin-7 / Q000378:metabolism; D016850:Limbus Corneae / Q000378:metabolism / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D055271:Mucin 5AC / Q000378:metabolism; D011446:Prospective Studies; D013234:Stem Cells / Q000378:metabolism / Q000473:pathology*; D055815:Young Adult
chemical_list: D015415:Biomarkers; D005136:Eye Proteins; C507494:KRT12 protein, human; C507426:KRT13 protein, human; C507497:KRT7 protein, human; D053551:Keratin-12; D053535:Keratin-13; D053539:Keratin-19; D053552:Keratin-7; C524678:MUC5AC protein, human; D055271:Mucin 5AC