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) 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/mm2. It was significantly higher in ocular cicatricial pemphigoid patients: 386 +/- 43 cells/mm2 (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.
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) 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/mm2. It was significantly higher in ocular cicatricial pemphigoid patients: 386 +/- 43 cells/mm2 (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.