Contributions
Abstract: P437
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Immunology
Introduction: Mesenchymal Stem Cells (MSCs) have dual effects: immunomodulatory and tissue protective and possible regenerative. Thus they have potential for cell-based therapies in MS. Previous studies show a differential ability of MSC to regulate Th17 and Th1 cells, but few studies have compared directly the immunomodulatory effects of MSC from MS patients and healthy volunteers towards autologous immune cells such as peripheral blood mononuclear cells (PBMC). In addition, MSC effects on T cells expressing GM-CSF are not known. In this study, we compared the immunomodulatory activity of MSC from patients with MS and those from healthy volunteers against autologous immune cells.
Objective: To compare the immunomodulatory activity of mesenchymal stem cells from patients with MS and healthy volunteers against autologous immune cells.
Aims: To gain insight into MSC in MS.
Methods: PBMC from 5 SPMS patients and 6 healthy controls (HC) were stimulated with anti-CD3/CD28 in monoculture or coculture with autologous bone marrow MSC (>95% MSC phenotype panel positive, passage 5 or 6) for 72h and 5h restimulation with PMA/ionomycin, followed by intracellular staining for IL-17, IFN-γ and GM-CSF. Cytokine expression was then quantified using flow cytometry. Supernatants of unstimulated MSC prior to co-culture were tested for a panel of cytokines and chemokines using a multiplex array and individual cytokine ELISA for some cytokines. Gene expression profiling was performed to explore differences between the MSC from MS and controls. For this, Lexogen QuantSeq library prep, which produces mostly 3' end near poly-A reads from mRNAs, was employed.
Results: Unstimulated MSC from MS patients produced less IL-10 (p=0.03) and more osteopontin (OPN) (p=0.002) than those from HC. MSC from HC reduced the proportion of autologous T cells expressing IL-17 (Th17), IFN-g (Th1), and GM-CSF by ~40%, whereas those from MS patients had the opposite effect by increasing autologous Th17 cells, GM-CSF-expressing T cells by ~100%, and did not suppress Th1 cells. Gene expression profiling shows down-regulation of pathways and regulators of TGF-β1.
MSC from SPMS patients have deficient immunomodulatory ability towards immune cells from the same patients, compared to MSC from HC in interaction with autologous immune cells. MSC trials are feasible but immunological monitoring is necessary. Restoration of immunomodulatory function (e.g. transplanting MSC with exogenous IL-10) may be helpful.
Disclosure: Supported by the MS Society of Great Britain and Northern Ireland and by a European Federation of Neurological Societies (now European Academy of Neurology) Fellowship.
Cris S. Constantinescu: nothing to disclose; Nanci Frakich: nothing to disclose, Sarah Thevathas: nothing to disclose; Radu Tanasescu: nothing to disclose; Bruno Gran: nothing to disclose; David Onion: nothing to disclose; Ian Spendlove: nothing to disclose; Philip Bath: nothing to disclose; Louise Showe: nothing to disclose; Andrew Kossenkov: nothing to disclose; Cherry Chang: nothing to disclose; Rhodri Jones: nothing to disclose
Abstract: P437
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Immunology
Introduction: Mesenchymal Stem Cells (MSCs) have dual effects: immunomodulatory and tissue protective and possible regenerative. Thus they have potential for cell-based therapies in MS. Previous studies show a differential ability of MSC to regulate Th17 and Th1 cells, but few studies have compared directly the immunomodulatory effects of MSC from MS patients and healthy volunteers towards autologous immune cells such as peripheral blood mononuclear cells (PBMC). In addition, MSC effects on T cells expressing GM-CSF are not known. In this study, we compared the immunomodulatory activity of MSC from patients with MS and those from healthy volunteers against autologous immune cells.
Objective: To compare the immunomodulatory activity of mesenchymal stem cells from patients with MS and healthy volunteers against autologous immune cells.
Aims: To gain insight into MSC in MS.
Methods: PBMC from 5 SPMS patients and 6 healthy controls (HC) were stimulated with anti-CD3/CD28 in monoculture or coculture with autologous bone marrow MSC (>95% MSC phenotype panel positive, passage 5 or 6) for 72h and 5h restimulation with PMA/ionomycin, followed by intracellular staining for IL-17, IFN-γ and GM-CSF. Cytokine expression was then quantified using flow cytometry. Supernatants of unstimulated MSC prior to co-culture were tested for a panel of cytokines and chemokines using a multiplex array and individual cytokine ELISA for some cytokines. Gene expression profiling was performed to explore differences between the MSC from MS and controls. For this, Lexogen QuantSeq library prep, which produces mostly 3' end near poly-A reads from mRNAs, was employed.
Results: Unstimulated MSC from MS patients produced less IL-10 (p=0.03) and more osteopontin (OPN) (p=0.002) than those from HC. MSC from HC reduced the proportion of autologous T cells expressing IL-17 (Th17), IFN-g (Th1), and GM-CSF by ~40%, whereas those from MS patients had the opposite effect by increasing autologous Th17 cells, GM-CSF-expressing T cells by ~100%, and did not suppress Th1 cells. Gene expression profiling shows down-regulation of pathways and regulators of TGF-β1.
MSC from SPMS patients have deficient immunomodulatory ability towards immune cells from the same patients, compared to MSC from HC in interaction with autologous immune cells. MSC trials are feasible but immunological monitoring is necessary. Restoration of immunomodulatory function (e.g. transplanting MSC with exogenous IL-10) may be helpful.
Disclosure: Supported by the MS Society of Great Britain and Northern Ireland and by a European Federation of Neurological Societies (now European Academy of Neurology) Fellowship.
Cris S. Constantinescu: nothing to disclose; Nanci Frakich: nothing to disclose, Sarah Thevathas: nothing to disclose; Radu Tanasescu: nothing to disclose; Bruno Gran: nothing to disclose; David Onion: nothing to disclose; Ian Spendlove: nothing to disclose; Philip Bath: nothing to disclose; Louise Showe: nothing to disclose; Andrew Kossenkov: nothing to disclose; Cherry Chang: nothing to disclose; Rhodri Jones: nothing to disclose