Limbal conjunctival Langerhans cell density in ocular cicatricial pemphigoid: an indirect immunofluorescence study on Dispase-split conjunctiva

Purpose. To evaluate limbal conjunctival Langerhans cell density in ocular cicatricial pemphigoid patients versus normal controls. Langerhans cells obviously play a major role in T-cell activation and are involved in corneal and conjunctival inflammatory diseases. Methods. We used a protease (Dispase II(R)) on inferior limbal conjunctival biopsies to separate the epithelium from the substantia propria and performed indirect immunofluorescence to analyze CD1a+ (a specific Langerhans cell surface antigen) cell density on flat-mounted epithelial sheets obtained from 30 normal controls and 11 patients presenting with ocular cicatricial pemphigoid. Results. This technique was quick and reproducible. The mean Langerhans cell density in normal limbal conjunctiva was 272+/-37 cells/mm(2). It was significantly higher in ocular cicatricial pemphigoid patients: 386+/-43 cells/mm(2) (p = 0.001). Conclusions. Conjunctival Langerhans cell density in ocular surface inflammatory diseases can best be evaluated by indirect immunofluorescence, following epithelial sheet separation from the substantia propria, using Dispase II(R).