M. Kouwenberg, L.B. Hilbrands, J. van der Vlag
Chair(s): dr. Michiel Betjes, Erasmus MC, Rotterdam
Thursday 10 march 2016
13:00 - 13:30h at Foyer
Parallel session: Postersessies XI - Opgesplitst in 3 tijdblokken en 3 categoriëen (klinisch, basaal, donatie)
Syndecan-1 is a heparan sulphate proteoglycan capable of binding chemokines and growth factors by its heparan sulphate side chains. Both dendritic cells (DCs) and T cells express syndecan-1. We previously demonstrated an immunomodulatory role of syndecan-1 in an experimental model of glomerulonephritis. A possible role of syndecan-1 in the interaction between DC and T cells, of paramount importance in the initiation of allograft rejection, has not been studied yet. In this study we aimed to investigate the role of syndecan-1 in DC - T cell interaction, as well as DC and T cell functioning.
Materials and methods:
The role of syndecan-1 on DC cell maturation was studied in vitro by incubation of mouse syndecan-1-/- or wild type (WT; C57Bl6) bone marrow derived DCs with different TLR ligands (PAM3CysSer, LPS, ODN1826). Maturation was analyzed by measuring the expression of co-stimulatory molecules (CD40, CD86) using flow cytometry. The role of syndecan-1 on T cells was studied in vitro by stimulating mouse syndecan-1-/- or wild type (WT; C57Bl6) CFSE labeled splenocytes either with concanavalin A or bone marrow derived Balb/c DCs. Proliferation was evaluated by dilution of CFSE signal. Co-culture supernatants were analyzed for cytokine levels to evaluate T cell differentiation.
Syndecan-1 deficient DCs stimulated with a TLR4 or TLR9 ligand showed a decreased co-stimulatory molecule expression compared to WT DCs. Syndecan-1 deficient T cells showed a diminished proliferative response upon low dose (0.25 µg/ml) concanavalin A and immature DC stimulation. Although the proliferative response upon mature DC and 0.5 µg/ml concanavalin A was comparable between groups, IL17 production was significantly reduced. Levels of IL2, IL4, IL10 and IFNγ in culture supernatant did not differ.
Syndecan-1 deficient DCs have a lower expression of co-stimulatory molecules after incubation with a TLR4 or TLR9 ligand, suggesting a role for syndecan-1 in DC maturation. Syndecan-1 seems also involved in T cell functioning, illustrated by diminished proliferative response upon stimulation by (immature) DC or (low dose) concanavalin A, and decreased IL17 production.