Co-expression of CD154 was used as a marker to identify activated antigen specific cytokine-secreting CD4+ cells [28]. specific treatment of autoimmune diseases. Introduction Studies around the prevention and treatment of autoimmune diseases have been focussed to the identification of antigens from pathogens and autoantigens responsible for triggering autoimmune reactions. Under decided conditions, autoantigens able to induce autoimmune disease can also suppress disease in a number of experimental models [1], [2]. Induction of antigen specific tolerance based on the persistence of antigen can be achieved after injection of high-dose antigen or repeated injections of low dose soluble antigens [3], [4], [5]. T cell deletion, anergy mechanisms and active suppression by regulatory cells constitute essential factors in the maintenance of tolerance induced by these means [3], [6]. Active suppression Celgosivir represents one of the dominant mechanisms in the control of autoreactivity characterized by deviation of the immune response via the secretion of suppressive cytokines [7], [8]. The use of Rabbit polyclonal to ZNF10 multivalent antigens represents an useful approach to deal with the dose/concentration of the antigen required to induce tolerance [9], [10], [11]. As an example, the synthetic repetitive Copolymer-1 (Glatiramer Acetate), approved as a therapy for relapsing remitting multiple sclerosis [12], contains sequences that cross react with the myelin basic protein [13], [14] and produce immunomodulatory effects involving the induction of specific T suppressor cells and bystander suppression mechanisms [15]. Despite several therapies for multiple sclerosis exist, their efficacy is very limited and most of the drugs slow Celgosivir the progression of the disease and reduce the quantity of relapses, however no total remedy is usually achieved [16], [17]. Our previous studies describe the role of repetitive oligomerized peptides in the control of autoimmune diseases. It was shown that a single low dose injection of oligomers, consisting of repeats of an encephalitogenic T cell epitope from your proteolipid protein of myelin, controlled the development of EAE in mice [9] and oligomers of the neuritogenic epitope of myelin P2 protein prevented the induction of experimental autoimmune neuritis (EAN) [10]. Similarly, multimerized self epitopes in the type I diabetes model demonstrated to provide protection against the disease and it was correlated with the growth of FoxP3+ regulatory cells [11]. Oligomers have proved to be effective in inducing strong immune response because of their ability to crosslink efficiently class II molecules of the major histocompatibilty complex (MHC-II) and to trigger signalling through the T cell receptor (TCR). This might result in improved antigenicity due to the activation of antigen presenting cells [18] and increased T cell proliferation [19]. The mechanisms of action of oligomerized peptides in suppressing the progression of autoimmune diseases are Celgosivir not completely elucidated. Their suppressive effect has been explained by the induction of anergy [20] or the growth of regulatory cells [11]. Some of the mechanisms underlying the tolerogenic capacity of repeat antigens are explained in this study. The ability of oligomer peptides made up of self-antigens to control the development as well as the progression of ongoing experimental autoimmune disease was correlated with the induction of protective tolerance mainly mediated by suppressive cytokines. Materials and Methods Ethics Statement The animals were maintained and dealt with according to the Directive 86/609/ECC of the European Community Council and to the institutional, state and federal guidelines. All animal experiments were approved by the Landesamt fr Arbeitsschutz, Gesundheitsschutz und Technische Sicherheit (Berlin, Germany). Animals were housed under standard conditions of 12-hour light/dark cycle and given access to food and water bacteria using recombinant techniques as explained [18]. In brief, double-stranded oligonucleotide models encoding the T cell epitopes of the PLP139C151 (C140S) oligomers were generated by annealing two complementary strands of synthetic oligonucleotides (PLP139C151 (C140S), +strand: using recombinant techniques as previously explained [18]. Endotoxin was removed from the polypeptide oligomers by separation on a reversed-phase C4-HPLC column (Vydac) and tested for endotoxin with the colorimetric limulus test (Charles River Laboratories). Antibodies For Celgosivir in injection, mouse monoclonal antibodies (mAb), were produced from hybridoma cell lines at the Maximum Delbrck Center. All hybridoma cell lines were kindly provided by Prof. Alexander Scheffold, Deutsches Rheuma-Forschungszentrum Berlin, Germany (DRFZ): anti-interleukin-10 (-IL-10) (clone JES5-2A5) [21], anti-IL-10 receptor (-IL-10R) (clone 1B1.3a) [22], anti-transforming growth factor beta (-TGF-) (clone 1D11) [23], anti-interleukin-4 (-IL4) (clone 11B11) [24], -CD25 (clone PC61) [25]. Rat IgG1 isotype (clone G1-113) was obtained from Sigma. Briefly, antibodies were harvested from culture supernatants and precipitated with ammonium sulphate. Afterwards they were purified over Protein G columns (Thermo scientific-Pierce, Germany) and dialysed against phosphate buffered saline (PBS). The protein concentration.