Effects of IL-4 and IL-12 on experimental immune-mediated blepharoconjunctivitis in Brown Norway rats

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Abstract
IL‐12 and IL‐4 are critical cytokines for Th1 and Th2 differentiation, respectively. To assess the roles of these cytokines in the development of experimental immune‐mediated blepharoconjunctivitis (EC) in Brown Norway (BN) rats, their effects were tested either in vitro or in vivo. Draining lymph node cells from rats immunized with ragweed pollen (RW) in Al(OH)3 were collected and cultured for 3 days with RW in the presence of IL‐4, IL‐12, or PBS as a control. After harvesting the culture supernatants for cytokine ELISA and the cells for cytokine reverse transcriptase‐polymerase chain reaction, 10 million cells were injected intravenously into syngeneic recipient rats (n = 12 per group). The rats were challenged with RW by eye drops 4 days after transfer. Eyes were harvested for histology 24 h later. Furthermore, IL‐12 (500 ng per injection) or PBS was injected intraperitoneally every other day seven times from the day of active immunization (n = 6 per group). One day after the last injection, rats were challenged and EC was evaluated as above. Transfer of cells with IL‐4 in vitro augmented eosinophilic infiltration in the conjunctiva compared with the other two groups, whereas IL‐12 in vitro suppressed eosinophilic infiltration and increased lymphocytic infiltration. Interferon‐gamma production was augmented by IL‐12. IL‐4 RNA expression was augmented by IL‐4. IL‐12 administration in vivo augmented lymphocytic infiltration in the conjunctiva without affecting infiltration of eosinophils. In conclusion, IL‐4 and IL‐12 either in vitro or in vivo augmented Th2 and Th1 immunity, respectively, thus leading to distinct histological features of EC.

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