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F. Baudouin, H. Liang, B. Dupas, C. Baudouin; Live Cell Trafficking in Rabbit Conjunctiva-Lymphoid Associated Tissue (calt) After Inflammatory and Toxic Stimuli. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4780.
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To study in vivo in rabbit eyes the cell trafficking occurring in the organized conjunctiva-associated lymphoid tissue (O-CALT) during conjunctival inflammation and toxic models.
In total, 45 rabbits were used: 30 for conjunctivitis model and 15 for toxic model. Adult male New Zealand albino rabbits were used in the following conjunctival challenge models: 30 animals were randomly divided into 5 groups and were subconjunctivally injected with sterile phosphate-buffered saline (PBS), lipopolysaccaride (LPS) or TNF, with or without a neutralizing antibody anti-TNF. Other 15 rabbits were used for toxicological models and were instilled with 50 µl of either: PBS, the commercial solutions of latanoprost, travoprost, bimatoprost or benzalkonium-free travoprost. All solutions were applied at 5-min intervals for a total of 15 times. By turning over the superior eyelid, the CALT patterns of superior conjunctiva were analyzed using a corneal in vivo confocal microscope (IVCM). Immunohistology was performed in whole mount conjunctiva and cryosections for detecting CD45+ lymphocytes.
We successfully observed in vivo aspects of the rabbit CALT. The conjunctivitis model induced by LPS was characterized by inflammatory cell infiltration in the upper and lower layers of CALT and cell circulation inside the lymph vessels. TNF alone induced moderate inflammatory infiltration in CALT. However anti-TNF antibodies could significantly decrease LPS/TNF-induced inflammation. CD45+ lymphocytes were strongly expressed in the CALT after injection of LPS or TNF at H4, and decreased with injection of anti-TNF. Antiglaucoma eye drops also stimulated the cell trafficking in the CALT, and seemed to be primarily related to the concentration of their common preservative benzalkonium chloride.
We showed for the first time the in vivo aspect of normal/pathological cell trafficking in rabbit O-CALT after inflammatory/toxic stimuli. IVCM analysis in CALT could be a pertinent tool in the future for the comprehension of ocular surface defense mechanisms, and the inflammatory cell trafficking in CALT could constitute new criteria for evaluating ocular surface inflammation and drug-induced damages.
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