Mesenchymal stem cells generate a CD4+CD25+Foxp3+ regulatory T cell population during the differentiation process of Th1 and Th17 cells

Abstract

IntroductionMesenchymal stem cells (MSCs) are adult, multipotent, stem cells with immunomodulatory properties. The mechanisms involved in the capacity of MSCs to inhibit the proliferation of proinflammatory T lymphocytes, which appear responsible for causing autoimmune disease, have yet to be fully elucidated. One of the underlying mechanisms studied recently is the ability of MSCs to generate T regulatory (Treg) cells in vitro and in vivo from activated peripheral blood mononuclear cells (PBMC), T-CD4+ and also T-CD8+ cells. In the present work we investigated the capacity of MSCs to generate Treg cells using T-CD4+ cells induced to differentiate toward the proinflammatory Th1 and Th17 lineages.MethodsMSCs were obtained from mouse bone marrow and characterized according to their surface antigen expression and their multilineage differentiation potential. CD4+ T cells isolated from mouse spleens were induced to differentiate into Th1 or Th17 cells and co-cultured with MSCs added at day 0, 2 or 4 of the differentiation processes. After six days, CD25, Foxp3, IL-17 and IFN-γ expression was assessed by flow cytometry and helios and neuropilin 1 mRNA levels were assessed by RT-qPCR. For the functional assays, the ‘conditioned’ subpopulation generated in the presence of MSCs was cultured with concanavalin A-activated CD4+ T cells labeled with carboxyfluorescein succinimidyl ester. Finally, we used the encephalomyelitis autoimmune diseases (EAE) mouse model, in which mice were injected with MSCs at day 18 and 30 after immunization. At day 50, the mice were euthanized and draining lymph nodes were extracted for Th1, Th17 and Treg detection by flow cytometry.ResultsMSCs were able to suppress the proliferation, activation and differentiation of CD4+ T cells induced to differentiate into Th1 and Th17 cells. This substantial suppressive effect was associated with an increase of the percentage of functional induced CD4+CD25+Foxp3+ regulatory T cells and IL-10 secretion. However, using mature Th1 or Th17 cells our results demonstrated that while MSCs suppress the proliferation and phenotype of mature Th1 and Th17 cells they did not generate Treg cells. Finally, we showed that the beneficial effect observed following MSC injection in an EAE mouse model was associated with the suppression of Th17 cells and an increase in the percentage of CD4+CD25+Foxp3+ T lymphocytes when administrated at early stages of the disease.ConclusionsThis study demonstrated that MSCs contribute to the generation of an immunosuppressive environment via the inhibition of proinflammatory T cells and the induction of T cells with a regulatory phenotype. Together, these results might have important clinical implications for inflammatory and autoimmune diseases.

Mesenchymal stem cells for the treatment of neurological diseases: Immunoregulation beyond neuroprotection

“…Treg cells in EAE [53]. [54] (reviewed in [55]), as well as, more importantly, by the therapeutic effect of B-cell depletion in MS [56].…”

Tissue Engineering Approaches to Modulate the Inflammatory Milieu following Spinal Cord Injury

“…MSCs and NSCs modulate the immune cells through numerous mechanisms, including direct cell-cell contact and indirect contact via cytokines and signaling molecules [Wang et al, 2009; Bonnamain et al, 2012a; English, 2013; Luz-Crawford et al, 2013; Barbeau et al, 2014; Gao et al, 2014; Nazmi et al, 2014; Obermajer et al, 2014; Yan et al, 2014]. NSCs release soluble factors such as TGF-β1, prostaglandin E, nitric oxide, and hemeoxygenase-1 that increase regulatory T cell populations at the expense of effector T cells [Wang et al, 2009; Bonnamain et al, 2012a; Nazmi et al, 2014].…”

Immunomodulatory Effects of Mesenchymal Stem Cells on T- and B-Cells in a Quiescent State in a Chronic Experimental Model of Autoimmune Encephalomyelitis

“…47 Finally, in a mouse model of EAE, it has been reported that MSC controlled the induction of T-cells with a regulatory phenotype and the inhibition of pro-inflammatory T-cells. 48 In the light of the foregoing data, we tested the therapeutic potential of intravenously injected BM-MSC from healthy donors in a chronic EAE model (Lewis 1A rats). We studied in vitro the immunomodulatory effects of MSC on the B-and T-lymphocytes of both control and EAE animals and the effects of intravenously administered MSC on EAE episodes.…”