Fingolimod attenuates splenocyte-induced demyelination in cerebellar slice cultures

PLoS One. 2014 Jun 9;9(6):e99444. doi: 10.1371/journal.pone.0099444. eCollection 2014.

Abstract

The family of sphingosine-1-phosphate receptors (S1PRs) is G-protein-coupled, comprised of subtypes S1PR1-S1PR5 and activated by the endogenous ligand S1P. The phosphorylated version of Fingolimod (pFTY720), an oral therapy for multiple sclerosis (MS), induces S1PR1 internalisation in T cells, subsequent insensitivity to S1P gradients and sequestering of these cells within lymphoid organs, thus limiting immune response. S1PRs are also expressed in neuronal and glial cells where pFTY720 is suggested to directly protect against lysolecithin-induced deficits in myelination state in organotypic cerebellar slices. Of note, the effect of pFTY720 on immune cells already migrated into the CNS, prior to treatment, has not been well established. We have previously found that organotypic slice cultures do contain immune cells, which, in principle, could also be regulated by pFTY720 to maintain levels of myelin. Here, a mouse organotypic cerebellar slice and splenocyte co-culture model was thus used to investigate the effects of pFTY720 on splenocyte-induced demyelination. Spleen cells isolated from myelin oligodendrocyte glycoprotein immunised mice (MOG-splenocytes) or from 2D2 transgenic mice (2D2-splenocytes) both induced demyelination when co-cultured with mouse organotypic cerebellar slices, to a similar extent as lysolecithin. As expected, in vivo treatment of MOG-immunised mice with FTY720 inhibited demyelination induced by MOG-splenocytes. Importantly, in vitro treatment of MOG- and 2D2-splenocytes with pFTY720 also attenuated demyelination caused by these cells. In addition, while in vitro treatment of 2D2-splenocytes with pFTY720 did not alter cell phenotype, pFTY720 inhibited the release of the pro-inflammatory cytokines such as interferon gamma (IFNγ) and interleukin 6 (IL6) from these cells. This work suggests that treatment of splenocytes by pFTY720 attenuates demyelination and reduces pro-inflammatory cytokine release, which likely contributes to enhanced myelination state induced by pFTY720 in organotypic cerebellar slices.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cerebellum / immunology*
  • Cerebellum / pathology*
  • Coculture Techniques
  • Cytokines / metabolism
  • Demyelinating Diseases / immunology*
  • Disease Models, Animal
  • Female
  • Fingolimod Hydrochloride
  • Immunosuppressive Agents / pharmacology*
  • Inflammation Mediators / metabolism
  • Mice
  • Mice, Transgenic
  • Myelin-Oligodendrocyte Glycoprotein / immunology
  • Phenotype
  • Propylene Glycols / pharmacology*
  • Sphingosine / analogs & derivatives*
  • Sphingosine / pharmacology
  • Spleen / cytology*
  • Spleen / immunology*
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism
  • Tissue Culture Techniques

Substances

  • Cytokines
  • Immunosuppressive Agents
  • Inflammation Mediators
  • Myelin-Oligodendrocyte Glycoprotein
  • Propylene Glycols
  • Fingolimod Hydrochloride
  • Sphingosine

Grants and funding

This work was supported in part by research grants from Trinity College Dublin Ireland and Novartis Pharma Basel Switerland. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Note, however, that AKM is an employee of Novartis Pharma Basel and had a role in study design, data collection and analysis, decision to publish and preparation of the manuscript. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials under relevant material transfer agreements.