| Autor: |
Rodi M; Laboratory of Immunohematology, Medical School, University of Patras, 26500 Patras, Greece., de Lastic AL; Laboratory of Immunohematology, Medical School, University of Patras, 26500 Patras, Greece., Panagoulias I; Laboratory of Immunohematology, Medical School, University of Patras, 26500 Patras, Greece., Aggeletopoulou I; Laboratory of Immunohematology, Medical School, University of Patras, 26500 Patras, Greece., Kelaidonis K; NewDrug P.C., Patras Science Park, 26504 Patras, Greece., Matsoukas J; NewDrug P.C., Patras Science Park, 26504 Patras, Greece.; Immunology and Translational Research, Institute for Health and Sport, Victoria University, Melbourne, VIC 3030, Australia.; Department of Physiology and Pharmacology, Cumming School of Medicine, University of Calgary, Calgary, AB T2N1N4, Canada.; Department of Chemistry, University of Patras, 26504 Patras, Greece., Apostolopoulos V; Immunology and Translational Research, Institute for Health and Sport, Victoria University, Melbourne, VIC 3030, Australia.; Immunology Program, Australian Institute for Musculoskeletal Science (AIMSS), Melbourne, VIC 3021, Australia., Mouzaki A; Laboratory of Immunohematology, Medical School, University of Patras, 26500 Patras, Greece. |
| Abstrakt: |
We have previously performed preclinical studies with the oxidized mannan-conjugated peptide MOG35-55 (OM-MOG35-55) in vivo (EAE mouse model) and in vitro (human peripheral blood) and demonstrated that OM-MOG35-55 suppresses antigen-specific T cell responses associated with autoimmune demyelination. Based on these results, we developed different types of dendritic cells (DCs) from the peripheral blood monocytes of patients with multiple sclerosis (MS) or healthy controls presenting OM-MOG35-55 or MOG-35-55 to autologous T cells to investigate the tolerogenic potential of OM-MOG35-55 for its possible use in MS therapy. To this end, monocytes were differentiated into different DC types in the presence of IL-4+GM-CSF ± dexamethasone (DEXA) ± vitamin D3 (VITD3). At the end of their differentiation, the DCs were loaded with peptides and co-cultured with T cells +IL-2 for 4 antigen presentation cycles. The phenotypes of the DC and T cell populations were analyzed using flow cytometry and the secreted cytokines using flow cytometry or ELISA. On day 8, the monocytes had converted into DCs expressing the typical markers of mature or immature phenotypes. Co-culture of T cells with all DC types for 4 antigen presentation cycles resulted in an increase in memory CD4+ T cells compared to memory CD8+ T cells and a suppressive shift in secreted cytokines, mainly due to increased TGF-β1 levels. The best tolerogenic effect was obtained when patient CD4+ T cells were co-cultured with VITD3-DCs presenting OM-MOG35-55, resulting in the highest levels of CD4+PD-1+ T cells and CD4+CD25+Foxp3+ Τ cells. In conclusion, the tolerance induction protocols presented in this work demonstrate that OM-MOG35-55 could form the basis for the development of personalized therapeutic vaccines or immunomodulatory treatments for MS. |
