To determine whether the ocular surface inflammation in uveitis mimics or counteracts intraocular inflammatory pathways by directly comparing T-helper (Th) lymphocytes Th1 and Th2 markers in conjunctival and ciliary body expression in endotoxin-induced uveitis (EIU). This study used the following inflammatory markers: chemokine receptor, CC chemokine receptor 4 (CCR4), and its ligand, macrophage-derived chemokine (MDC), to evaluate Th2 participation; chemokine receptor, CCR5, to evaluate the Th1 system; and its ligand, regulated on activation normal T cell expressed and secreted (RANTES), to evaluate both Th1 and Th2 systems.
Immunohistochemistry and real-time polymerase chain reaction (RT–PCR) were used to compare protein and RNA expression of CCR4, MDC, CCR5, and RANTES in the conjunctiva and ciliary body in EIU 6 h and 24 h after the lipopolysaccharide (LPS) injection and in control (without injection) Lewis rats.
Immunohistochemistry with CCR5, RANTES, and MDC showed an increase in fluorescent staining in the conjunctiva and ciliary body in the rats with uveitis compared to the control rats. For CCR4, immunostaining was comparable in the conjunctiva and ciliary body and did not show any clear differences between control rats and rats with EIU. For RANTES, MDC, and CCR5, RT-PCR showed a significantly higher RNA expression in conjunctiva and in ciliary body at 6 h compared to 24 h and controls. For CCR4, RT-PCR did not illustrate any significant differences in conjunctiva and in ciliary body between all groups of animals.
Protein and RNA expressions of RANTES, MDC, and CCR5 were higher in EIU rats than in control rats in the conjunctiva and ciliary body whereas the CCR4 level was not modified in the conjunctiva and ciliary body of EIU rats when compared to controls. Th1 activation seemed to predominate in this model with high levels of CCR5 expression and no increased expression of CCR4, but Th2 participation with MDC was noted. The expression of RANTES, MDC, CCR4 and CCR5 in EIU was quite similar between the conjunctiva and the ciliary body, so conjunctival inflammation might reproduce the intraocular inflammation, probably generated by local extension and diffusion in this model. If the ocular surface mimics intraocular inflammatory pathways, the conjunctiva may provide a new and easier access for uveitis studies.