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Abstract #9640 Published in IGR 5-3
Flow cytometric analysis of HLA-DR, IL-6 and IL-8 expression by conjunctival epithelial cells from patients with prolonged topical antiglaucoma treatments
Bensoussan L; Blondin C; Baudouin C; Hamard P; Sabeh Afaki G; Creuzot-Garcher C; Warnet JM; Brignole-Baudouin FJournal Français d'Ophtalmologie 2003; 26: 782-789
AIM: To investigate the expression of three inflammation markers, HLA-DR, IL-6, and IL-8, by conjunctival epithelial cells obtained using impression cytology (IC) samples from long-term treated glaucoma patients. PATIENTS AND METHODS: IC samples were obtained from the 60 following individuals: 45 patients suffering from primary open-angle glaucoma and receiving topical treatments for at least one year, and 15 subjects with no ophthalmological disease (controls). Membrane expression of HLA-DR and intracellular expression of IL-6 and IL-8 were quantified by indirect and direct immunofluorescence techniques, respectively. Fluorescence levels were quantified using calibrated fluorescent beads. RESULTS: The percentage of HLA-DR-positive cells was significantly higher on IC samples from multi-treated glaucoma patients and from patients treated by either preserved beta blocker or preserved prostaglandin analogue than on IC samples from control individuals. Interestingly, the percentage of HLA-DR-positive cells was not significantly increased upon treatment with unpreserved beta blocker. However, the percentage of IL-6- and IL-8-positive cells, as well as IL-6 and IL-8 expression levels, was significantly higher in patients than in controls, regardless of the treatment type and the presence of preservative. A significant positive correlation was found between HLA-DR and cytoplasmic IL-6 expression, HLA-DR and cytoplasmic IL-8, as well as between IL-6 and IL-8 cytoplasmic expressions. CONCLUSIONS: The present study confirms that there is an increased expression of HLA-DR in treated glaucoma patients compared to controls and demonstrates that antiglaucoma treatments lead to increased IL-6 and IL-8 expressions. It also indicates that benzalkonium-preserved eye drops and preserved multitherapy may induce stronger inflammatory responses than do unpreserved eye drops, although further studies are needed to determine the respective inflammatory role of preservative and therapeutic molecules. In this study, flow cytometry was used to detect the intracellular pro-inflammatory cytokines in conjunctival cells obtained by impression cytology. This standardized and reliable technique was a useful tool for assessing inflammatory and allergic ocular surface disorders. LA: French
Dr. L. Bensoussan, Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, EA 3123 Universite Pierre et Marie Curie, Paris 6, France
Classification:
2.1 Conjunctiva (Part of: 2 Anatomical structures in glaucoma)