Abstract
Purpose: :
To investigate the functional roles of TAM receptor tyrosine kinases in development of the ocular autoimmune diseases.
Methods: :
Circulating autoantibodies against IRBP1-20 were measured in a 1:200 and 1:100 dilution of sera. EAU was induced by active immunization with retinal specific antigen, IRBP1-20 in CFA. T cell proliferation assays were performed on the T cells isolated from the IRBP1-20-immunized wild-type or TAM knockout mice. The enriched T cells were seeded at 4x105 cells/well in 96-well plates containing the irradiated syngeneic splenocytes (1x105), and cultured at 37oC for 48 h in a total volume of 200ul medium with or without IRBP1-20 and IRBP161-180. Incorporation of the [3H]-thymidine was assessed using a microplate scintillation counter. Flow cytometry was done on a BD FACACalibur, using fluorescent Abs from BD or eBioscience. Real-time qPCR was performed on Stratagen MX3005.
Results: :
The TAM triple mutants displayed elevated levels of circulating autoantibodies, including those against the ocular autoantigens and eventually developed manifestation of human uveitis autoimmunity. These triply mutant mice exhibited higher sensitivity to immunization by the ocular autoantigens, e.g., Interphotoreceptor Retinoid-Binding Protein (IRBP). The splenocytes derived from those mice are more potent to activate ocular antigen-specific syngeneic T-cell proliferation. The T-cells isolated from mutants were more potent to cause inflammation in the host animals.
Conclusions: :
TAM triple mutant mice develop ocular autoimmune disorder and provide a new mouse model for studying the uveitis development and testing therapeutic methodologies.
Keywords: autoimmune disease • uveitis-clinical/animal model • retina