Contributions
Abstract: EP1496
Type: ePoster
Abstract Category: Pathology and pathogenesis of MS - 15 Immunology
Background: Multiple Sclerosis (MS) is a chronic inflammatory disease of the central nerve system. Multiple types of immune cells have been associated with MS aetiology, including monocytes, B cell and T helper (Th) cell subsets. The potential interaction between peripheral immune cells, Vitamin D and MS could affect the pathogenic mechanisms of MS and is the focus of this study.
Objectives: To investigate the association between peripheral immune cell responses, Vitamin D and MS ex vivo and in vitro in a case-control study.
Methods: We collected peripheral blood mononuclear cells (PBMCs) from 62 people with MS and 12 healthy controls (HC), and determined the proportion of immune cell subsets via Flow cytometry ex vivo. Furthermore, we stimulated isolated PBMCs with ConA (10µg/ml), ConA + 25(OH)D3 (500nM) or ConA + 1,25(OH)2D3 (100nM) for 96h in vitro. The Th cell subsets were analysed via Flow cytometry and the titer of cytokines in supernatants was determined via Cytometric Bead Array. The genotypes of Vitamin D-associated MS risk-SNPs (rs12368653, rs703842, rs2248359) were determined by 3'mismatch PCR. Data were compared using nonparametric tests (i.e., Mann-Whitney, Kruskal-Wallis and Friedman test).
Results: Ex vivo, the proportion of classic monocytes was decreased in monocytes (p=0.0144), while the proportion of B cell was increased in PBMCs (p=0.0089) in the MS cohort than in HC. In in vitro, for the whole samples, both 25(OH)D3 and 1,25(OH)2D3 significantly inhibited the production of IL-6 (p< 0.01), IL-10 (p< 0.001), TNF (p< 0.001), IFN-gamma (p< 0.001) and IL-17A (p< 0.001), and promoted the production of IL-4 (p< 0.01) by PBMCs after 96hrs of stimulation with ConA. Interestingly the HI PBMCs produced more IL-6 (p=0.0171), TNF (p=0.0001) and Th1 (p=0.0231) after stimulation with ConA for 96h than the MS group. The rs703842 risk allele AA was associated with significantly reduced production of IL-2 at 96h (p=0.0076). For the whole samples, the rs2248359 risk allele TT was associated with an increased proportion of Th17 and Th1 cell populations within the CD4+ T cell subset ex vivo and the Th1 population after 96h ConA stimulation (in vitro).
Conclusions: Peripheral immune cell variations and inflammatory cytokine profiles are influenced by Vitamin D and the MS status ex vivo and in vitro supporting a potential role for vitamin D in the pathogenesis of MS.
Keywords: peripheral immune cell subsets, Vitamin D, cytokines.
Disclosure:
Ming Lu: nothing to disclose
Bruce Taylor: nothing to disclose
Heinrich Korner: nothing to disclose
Funding: MS Research Austalia and Royal Hobart Research Foundation
Abstract: EP1496
Type: ePoster
Abstract Category: Pathology and pathogenesis of MS - 15 Immunology
Background: Multiple Sclerosis (MS) is a chronic inflammatory disease of the central nerve system. Multiple types of immune cells have been associated with MS aetiology, including monocytes, B cell and T helper (Th) cell subsets. The potential interaction between peripheral immune cells, Vitamin D and MS could affect the pathogenic mechanisms of MS and is the focus of this study.
Objectives: To investigate the association between peripheral immune cell responses, Vitamin D and MS ex vivo and in vitro in a case-control study.
Methods: We collected peripheral blood mononuclear cells (PBMCs) from 62 people with MS and 12 healthy controls (HC), and determined the proportion of immune cell subsets via Flow cytometry ex vivo. Furthermore, we stimulated isolated PBMCs with ConA (10µg/ml), ConA + 25(OH)D3 (500nM) or ConA + 1,25(OH)2D3 (100nM) for 96h in vitro. The Th cell subsets were analysed via Flow cytometry and the titer of cytokines in supernatants was determined via Cytometric Bead Array. The genotypes of Vitamin D-associated MS risk-SNPs (rs12368653, rs703842, rs2248359) were determined by 3'mismatch PCR. Data were compared using nonparametric tests (i.e., Mann-Whitney, Kruskal-Wallis and Friedman test).
Results: Ex vivo, the proportion of classic monocytes was decreased in monocytes (p=0.0144), while the proportion of B cell was increased in PBMCs (p=0.0089) in the MS cohort than in HC. In in vitro, for the whole samples, both 25(OH)D3 and 1,25(OH)2D3 significantly inhibited the production of IL-6 (p< 0.01), IL-10 (p< 0.001), TNF (p< 0.001), IFN-gamma (p< 0.001) and IL-17A (p< 0.001), and promoted the production of IL-4 (p< 0.01) by PBMCs after 96hrs of stimulation with ConA. Interestingly the HI PBMCs produced more IL-6 (p=0.0171), TNF (p=0.0001) and Th1 (p=0.0231) after stimulation with ConA for 96h than the MS group. The rs703842 risk allele AA was associated with significantly reduced production of IL-2 at 96h (p=0.0076). For the whole samples, the rs2248359 risk allele TT was associated with an increased proportion of Th17 and Th1 cell populations within the CD4+ T cell subset ex vivo and the Th1 population after 96h ConA stimulation (in vitro).
Conclusions: Peripheral immune cell variations and inflammatory cytokine profiles are influenced by Vitamin D and the MS status ex vivo and in vitro supporting a potential role for vitamin D in the pathogenesis of MS.
Keywords: peripheral immune cell subsets, Vitamin D, cytokines.
Disclosure:
Ming Lu: nothing to disclose
Bruce Taylor: nothing to disclose
Heinrich Korner: nothing to disclose
Funding: MS Research Austalia and Royal Hobart Research Foundation