F.19. Neural Stem Cells Expand FoxP3+ Treg In Vitro

June 29, 2017 | Autor: Samia Khoury | Categoria: Immunology, Clinical immunology
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Abstracts natural killer cells and mast cells. The role of Itk in T cell development and signaling has been elucidated by the generation of Itk-deficient mice which show impairments in CD4+ T cell development and reduced differentiation of T cells into the Th2 lineage. Functionally, the lack of Itk leads to reductions in Ca2+ mobilization, cytokine production, and proliferation following engagement of the T cell receptor. Itk-deficient mice also show reduced responsiveness to parasitic infections and diminished symptoms of allergic asthma in response to challenge with the allergen ovalbumin. Here we report on the role of Itk in the myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE) model, a murine model for the autoimmune disease multiple sclerosis (MS). In addition to delayed onset of disease, Itk-deficient mice had significantly reduced incidence of EAE and decreased clinical severity of disease versus wild-type controls. T cells from Itk-deficient mice immunized with MOG proliferated less and secreted lower levels of IFN-γ and TNF-α than T cells from wild type mice upon restimulation in vitro. Microscopically, Itk-deficient mice had a reduced number of inflammatory foci in the brain and spinal cord versus wildtype mice. The extent of axonal demyelination was also reduced in Itk-deficient mice. These data point to a role for Itk in the development and progression of EAE and to the therapeutic potential of Itk inhibition in inflammatory conditions such as MS. doi:10.1016/j.clim.2008.03.130

F.19. Neural Stem Cells Expand FoxP3+ Treg In Vitro Yue Wang, Bing Zhu, Wassim Elyaman, Jaime Imitola, Samia Khoury. Brigham and Women's Hospital, Boston, MA Endogenous neural stem cells (NSCs) are considered potential source for neural repair and regeneration in adult central nervous system. NSC transplantation has been shown to have therapeutic effect in experimental autoimmune encephalomyelitis (EAE), and the transplanted NSCs appear to have immunoregulatory functions. However, the mechanisms for how T cells are regulated are not very clear. The purpose of this study is to examine the regulatory effect of NSC during T cell activation in vitro. Methods: NSCs were isolated from the forebrains of E14.5 C57Bl/6 embryos and were cultured in FGF/EGF containing medium to maintain their self-renewing capacity and multipotency. We sorted splenic FoxP3-GFP - CD4+ T cells from the FoxP3-GFP B6 knock-in mice by flow cytometry. The T cells and the NSCs were co-cultured at a ratio of 4:1 in 48-well plate with plate-bound anti-CD3/anti-CD28 activation for 3-5 days. Results: 13.18 ± 7.03% of the FoxP3-GFP-CD4+ T cells in the NSC/T co-culture converted to FoxP3-GFP+CD4+ T cells, compared to 0.93 ± 0.72% in the stimulated T cells condition. Th1 and Th2 cytokines were both suppressed in the co-culture. NSCs were able to expand FoxP3-GFP+CD4+ regulatory T cells in the coculture as measured by BrdU-uptake. The NSC-conditioned media have similar effect as the co-culture. The Treg cells generated in the co-culture are functionally active and

S49 suppress T cell proliferation in vitro. Conclusions: Our data suggest that generation and expansion of Treg cells may be one of the mechanisms by which NSCs regulate T cell response in vivo. doi:10.1016/j.clim.2008.03.131

F.20. A c-jun N-terminal Kinase (JNK)/Apoptosis/ Inflammation-based Gene Card for Multiple Sclerosis Diagnosis and Patient Stratification Chiara Ferrandi, Fabien Richard, Patrizia Tavano, Valerie Barbié, Maurizio Mariani, Ehud Hauben, Paola Francesca Zaratin. Merck Serono Research/RBM S.p.A., Colleretto Giacosa (TO), Italy Clonal deletion of T cells with high affinities for selfpeptide-MHC via apoptosis is an essential mechanism leading to self-tolerance. Disruption of these mechanisms may lead to the development of autoimmune diseases. Accordingly, there is evidence that defective apoptosis of activated T lymphocytes is involved in the pathogenesis of MS. The c-jun N-terminal Kinase (JNKs) pathway is involved in cellular survival and apoptosis. To investigate the link between JNK, apoptosis and inflammatory pathways in relapsing-remitting MS (RRMS) we purified peripheral blood CD4+ and CD8+ T cells, as well as CD11b+ monocytes from RRMS patients in active phase (n = 7) and healthy volunteers (HV, n = 11). These cell subsets were cultured for 24 h with AS602801, a JNK non-isoform-specific inhibitor. Cytokine secretion and gene expression profiles were determined by cytokine bead array and quantitative PCR, respectively. In RRMS CD4+ T cells AS602801 significantly inhibited the expression of the apoptosis-related genes APAF1, DUSP6 and NUMB. In addition, it inhibited expression of the proinflammatory genes ITGA4 and IL1B. In RRMS CD8+ T cells, AS602801 increased IFN-γ protein release and elevated CD69 and GADD45B gene expression. In RRMS CD11b+ monocytes AS602801 inhibited production of IL-12p70 and IL-1β proteins and gene expression of IL27. It also enhanced the expression of SOCS1, CD40, NFKB, PDL1, CD80 and GADD45A. Overall, our findings suggest that AS602801 regulates CD4 and CD8 T cell apoptosis through modulation of antigen presenting cells phenotype and function. Moreover, the above data support the design of a JNK/Apoptosis/Inflammation gene card as diagnostic and/or prognostic integrated biomarker of MS pathology. (C.F. and F.A.R. equally contributed to the work). doi:10.1016/j.clim.2008.03.132

F.21. Immune Regulation of Experimental Autoimmune Encephalomyelitis (EAE) by RIP2-dependent Modulation of NF-κB Signaling Todd Fairhead,1 Rafael Fenutria,2 Michelle McCully,1 Caitlin Lemke,1 Brianne Davis,1 Francisco Lozano,2 Gill Strejan,1 Joaquin Madrenas.1 1Robarts Research Institute, University of Western Ontario, London, ON, Canada; 2Universitat de Barcelona, Barcelona, Spain

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