T cell vaccination induces the elimination of EAE effector T cells: Analysis using GFP-transduced, encephalitogenic T cells q Ilan Volovitz a, b, * , Yotvat Marmor a , Felix Mor a , Alexander Flügel c, d , Francesca Odoardi c, d , Lea Eisenbach a , Irun R. Cohen a a The Department of Immunology, The Weizmann Institute of Science, Rehovot 76100, Israel b The Department of Neurosurgery, The Tel-Aviv Sourasky Medical center, Tel-Aviv 64239, Israel c Max Planck Institute for Neurobiology, Martinsried 82152, Germany d Department of Neuroimmunology, Institute for Multiple Sclerosis Research, Gemeinnützige Hertie-Stiftung and University Medical Centre Göttingen, Göttingen 37073, Germany article info Article history: Received 21 April 2010 Received in revised form 25 May 2010 Accepted 28 May 2010 Keywords: Rodent e rat Experimental autoimmune encephalomyelitis e EAE Multiple sclerosis e MS T cell vaccination e TCV T cells Cell tracking methods abstract T cell vaccination (TCV) with irradiated encephalitogenic T cells induces resistance to EAE. However, the fate of the encephalitogenic T cells in vivo following TCV has yet to be studied. Here we used anti-MBP encephalitogenic T cells that were transduced to express GFP to study the effects of TCV on these cells. In naïve rats or in control-vaccinated (Ova-GFP) rats injected i.v. with GFP-labeled effector cells, high numbers of effector T cells were found along with macrophages, CD8 T cells and Non-GFP CD4 cells in the spleens, parathymic lymph nodes (PTLN) and spinal cords. In contrast, the recipients that had been treated with TCV (anti-MBP T-cell lines) showed few if any GFP-labeled effector T cells throughout the disease (day 1e8) and their spinal cords were almost clear of macrophages, CD4 and CD8 cells. Splenocytes in the control groups secreted IFNg in response to MBP and showed high numbers of IFNg secreting CD4 and CD8 cells in their spinal cords at the disease peak. In the TCV-protected groups, splenocytes showed no reactivity to MBP but secreted IFNg in response to irradiated encephalitogenic T cells e an anti-idiotypic response. Thus, TCV leads to a marked decrease in the numbers of effector T cells in the CNS and lymphoid organs, to a marked reduction in the Th1 cytokine producing cells in the CNS, and to the appearance of T cells responsive to the anti-MBP effector T cells. Ó 2010 Elsevier Ltd. All rights reserved. 1. Introduction Experimental autoimmune encephalomyelitis (EAE) serves as a prototype for T cell-mediated autoimmune diseases [1] and has been used as a model for the pathogenesis of multiple sclerosis (MS) to help identify potential therapeutic candidates for this disease [2]. One such therapeutic modality is vaccination with antigen-activated or mitogen-activated attenuated encephalitogenic T cells called T cell vaccination (TCV) [3,4]. TCV is used to activate the immune system to recognize and neutralize disease-inducing effector T cells [5,6]. Effective treatment using TCV [4] was demonstrated in various experimental autoimmune diseases in rodents such as in adjuvant arthritis [7], experimental autoimmune thyroiditis [8], collagen- induced arthritis [9], experimental autoimmune uveitis [10], lupus [11], and type-1 diabetes [12]. TCV has been applied successfully to humans to treat MS [4,5], rheumatoid arthritis [13], and lupus [14]. Thus, the protective mechanisms induced by TCV have clinical relevance. TCV was shown to activate anti-idiotypic T cells that are able to recognize the encephalitogenic T cells by unique TCR-peptide sequences [12,15]. Regulatory anti-idiotypic CD4 T cells [16] were able to shift the response from pathologic anti-myelin Th 1 responses to protective Th 2 responses [17] and to provide help to anti-idiotypic CD8 T cells, which appear to be the major suppressors of the autoimmune effector T cells [18]. CD8 þ suppressive T cells, induced following TCV in rats, were shown to specifically suppress the response of anti-MBP T cells to their cognate antigen [15], and to lyse them [18]. The CD8 regulator cells were reported to recognize TCR peptides presented on the MHC class Ib molecule, Qa-1, expressed on acti- vated CD4 T cells [19,20]. In human multiple sclerosis patients, CD8 Abbreviations: ATCC, American Type Culture Collection; NRS, Normal Rat Serum; RBC, Red Blood Cells; TCGF, Tcell Growth Factor; TBI, Total Body Irradiation; Ova, Ovalbumin; EAE, experimental autoimmune encephalomyelitis; TCV, T-cell vaccination; MBP/GBP, guinea-pig MBP (myelin basic protein); RBP, rat MBP; BBP, Bovine MBP; HBP, Human MBP. q All work was performed in the Weizmann Institute, Israel. * Corresponding author at: The Department of Immunology, Weizmann Institute of Science, Rehovot 76100, Israel. Tel.: þ972 8 934 3419; fax: þ972 8 9344103. E-mail address: Volovitz@yahoo.com (I. Volovitz). Contents lists available at ScienceDirect Journal of Autoimmunity journal homepage: www.elsevier.com/locate/jautimm 0896-8411/$ e see front matter Ó 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.jaut.2010.05.003 Journal of Autoimmunity 35 (2010) 135e144