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International Immunology Advance Access originally published online on September 6, 2004
International Immunology 2004 16(11):1573-1582; doi:10.1093/intimm/dxh158
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© 2004 The Japanese Society for Immunology

The fate of autoreactive, GFP+ T cells in rat models of uveitis analyzed by intravital fluorescence microscopy and FACS

Stephan R. Thurau1, Thorsten R. Mempel2,4, Alexander Flügel3, Maria Diedrichs-Möhring1, Fritz Krombach2, Naoto Kawakami3 and Gerhild Wildner1

1 Section of Immunobiology, Department of Ophthalmology and 2 Institute for Surgical Research, Ludwig-Maximilians-University, Munich and 3 Department of Neuroimmunology, Max-Planck-Institute of Neurobiology, Martinsried, Germany
4 Present address: Center for Blood Research and Department of Pathology, Harvard Medical School, Boston, USA

Correspondence to: S. R. Thurau; E-mail: Stephan.Thurau{at}med.uni-muenchen.de

Experimental autoimmune uveitis (EAU) is an inflammatory disease of the immune privileged inner eye, mediated by CD4+ Th1 cells specific for retinal autoantigens. To elucidate the fate of the T cells in the eye we adoptively transferred green fluorescent protein-positive (GFP+) T cells with specificity for R14, a peptide from interphotoreceptor retinoid-binding protein (IRBP) or OVA as foreign control antigen to naive Lewis rats. We also used the model of immunogenic uveitis, an inflammatory eye disease induced by intraocular injection of soluble OVA 1 day post transfer of OVA-specific GFP+ cells. We investigated the timing of ocular T cell infiltration and their immunological activation state by intravital fluorescence microscopy (IVFM) of the iris until onset of intraocular inflammation. Within 30 min of injection, GFP+ cells invaded the iris tissue, irrespective of their antigen specificity, whereas intraocular inflammation was only observed 3 days later, if cells recognized their respective antigen (R14-specific cells in EAU, OVA-specific cells in immunogenic uveitis). Using FACS analysis we found that activation markers were upregulated only on cells from uveitic eyes, but not from other sources, suggesting that intraocularly presented specific antigen is a prerequisite for T cell reactivation and subsequent recruitment of inflammatory cells.

Keywords: activation marker, experimental autoimmune uveitis, immunogenic uveitis, interphotoreceptor retinoid binding protein, T cell migration

Transmitting editor: T. Hünig


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