****Cheng-Rong Yu prepared this manuscript and should have been the first author.
Expression of IL-17 in mouse PBMC, lymph node and retina is temporally correlated with progression of experimental autoimmune uveoretinitis (EAU)
Uveitis is a diverse group of intraocular inflammatory diseases that cause severe visual loss and morbidity and is characterized by inflammatory attack of the uvea as well as the neuroretina 1. The disease may be of infectious or putative autoimmune etiology. The latter include birdshot retinochoroidopathy, sympathetic ophthalmia, Behçet’s disease, Vogt-Koyanagi-Harada syndrome, and ocular sarcoidosis 1. Understanding the immunopathogenic mechanisms of uveitis has benefited enormously by the development of an animal model of uveitis, experimental autoimmune uveitis (EAU). EAU is a predominantly a T cell-mediated intraocular inflammatory disease induced in susceptible species by active immunization with ocular-specific proteins (or peptides derived from them) and is transferable to naive syngeneic animals by injection of in vitro-activated CD4+, MHC class II-restricted T cell lines specific to retinal Ags 2-4. The two major uveitogenic retinal proteins are S-Ag (also termed “arrestin”) and interphotoreceptor retinoid-binding protein (IRBP) 2-4. In this study, we have used the EAU mouse model to further establish that TH17 cells are involved in pathogenic mechanisms of uveitis by showing that EAU can be ameliorated by treatment with antibody to IL-17.
The procedure consists of the following steps:
Active induction of experimental autoimmune uveoretinitis (EAU)
Preparation of antigen/CFA emulsion
Clinical grading of mice with EAU by fundoscopy
Preparation of sections for ocular histology
Inhibition of EAU by IL-17 neutralizing antibody
IL-27 expression in PBMC or retina is temporally correlated with progression of EAU
Other protocols related to our Nature Medicine paper can be found here:
"Detection of TH17 cells in human blood":http://www.natureprotocols.com/2007/08/30/th17_cells_contribute_to_uveit.php
"Western Blot Analysis":http://www.natureprotocols.com/2007/08/30/th17_cells_contribute_to_uveit_2.php
"Analysis by confocal microscopy":http://www.natureprotocols.com/2007/08/31/th17_cells_contribute_to_uveit_3.php
"Retinal cell isolation":http://www.natureprotocols.com/2007/08/31/th17_cells_contribute_to_uveit_5.php