Contributions
Abstract: P1148
Type: Poster
Abstract Category: Therapy - disease modifying - 26 Immunomodulation/Immunosuppression
Background: In the pursuit of re-establishing tolerance in multiple sclerosis, the use of tolerogenic dendritic cells (tolDC) is a promising strategy. For antigen loading, electroporation with mRNA encoding full-length proteins has the advantage of inducing presentation of all potentially relevant epitopes in a non-HLA-restricted manner. Hereby, this technique offers the potential to prevent epitope spreading and its subsequent negative effect on the clinical disease course.
Aim: To generate murine tolerance-inducing DC presenting myelin oligodendrocyte glycoprotein (MOG) by mRNA electroporation.
Materials and methods: 1α,25-dihydroxyvitamin D3-treated, bone marrow-derived tolDC were electroporated with MOG mRNA. The MOG-presenting capacity of electroporated mature control DC and tolDC was assessed with IFN-γ ELISPOT analysis following coculture with MOG-reactive splenocytes from experimental autoimmune encephalomyelitis (EAE) mice. The stability of the tolerogenic properties following mRNA electroporation was evaluated by assessing the cytokine secretion profile and
T cell-stimulatory capacity. MOG-specific proliferation of splenocytes from EAE mice treated with electroporated tolDC was evaluated with a thymidine incorporation assay.
Results: TolDC demonstrate low secretion of IL-12p70 and low stimulatory capacity in an allogeneic mixed lymphocyte reaction following activation with a pro-inflammatory stimulus. This maturation-resistant phenotype was not affected by mRNA electroporation. Moreover, MOG mRNA-electroporated tolDC displayed a marked suppressive effect on MOG-reactive splenocytes, as evidenced by a 86,6±5,0% reduction on average of IFN-γ-producing cells as compared to splenocytes stimulated with electroporated control DC. Finally, following in vivo administration of MOG mRNA-electroporated tolDC in EAE mice, splenocytes and lymph node cells demonstrated a reduction in MOG-specific proliferation compared with PBS-treated mice.
Conclusion: Electroporation of tolDC with MOG mRNA is an effective tool to generate MOG-presenting murine tolDC without affecting their tolerogenic properties. Moreover, MOG mRNA-electroporated tolDC effectively suppress EAE splenocytes in vitro and in vivo. These findings hold promise for further (pre)clinical translation. In this perspective, the clinical effect of MOG mRNA-electroporated tolDC treatment is currently being investigated in EAE mice.
Disclosure: This work was supported by a BOF-GOA Grant from the University of Antwerp, Belgium.
Judith Derdelinckx: holds a PhD Fellowship of the Research Foundation - Flanders (FWO), further nothing to disclose.
María José Mansilla: nothing to disclose.
Maxime De Laere: nothing to disclose.
Wai-Ping Lee: nothing to disclose.
Juan Navarro Barriuso: nothing to disclose.
Patrick Cras: nothing to disclose.
Barbara Willekens: nothing to disclose.
Peter Ponsaerts: nothing to disclose.
Zwi N. Berneman: nothing to disclose.
Eva María Martínez-Cáceres: nothing to disclose.
Nathalie Cools: nothing to disclose.
Abstract: P1148
Type: Poster
Abstract Category: Therapy - disease modifying - 26 Immunomodulation/Immunosuppression
Background: In the pursuit of re-establishing tolerance in multiple sclerosis, the use of tolerogenic dendritic cells (tolDC) is a promising strategy. For antigen loading, electroporation with mRNA encoding full-length proteins has the advantage of inducing presentation of all potentially relevant epitopes in a non-HLA-restricted manner. Hereby, this technique offers the potential to prevent epitope spreading and its subsequent negative effect on the clinical disease course.
Aim: To generate murine tolerance-inducing DC presenting myelin oligodendrocyte glycoprotein (MOG) by mRNA electroporation.
Materials and methods: 1α,25-dihydroxyvitamin D3-treated, bone marrow-derived tolDC were electroporated with MOG mRNA. The MOG-presenting capacity of electroporated mature control DC and tolDC was assessed with IFN-γ ELISPOT analysis following coculture with MOG-reactive splenocytes from experimental autoimmune encephalomyelitis (EAE) mice. The stability of the tolerogenic properties following mRNA electroporation was evaluated by assessing the cytokine secretion profile and
T cell-stimulatory capacity. MOG-specific proliferation of splenocytes from EAE mice treated with electroporated tolDC was evaluated with a thymidine incorporation assay.
Results: TolDC demonstrate low secretion of IL-12p70 and low stimulatory capacity in an allogeneic mixed lymphocyte reaction following activation with a pro-inflammatory stimulus. This maturation-resistant phenotype was not affected by mRNA electroporation. Moreover, MOG mRNA-electroporated tolDC displayed a marked suppressive effect on MOG-reactive splenocytes, as evidenced by a 86,6±5,0% reduction on average of IFN-γ-producing cells as compared to splenocytes stimulated with electroporated control DC. Finally, following in vivo administration of MOG mRNA-electroporated tolDC in EAE mice, splenocytes and lymph node cells demonstrated a reduction in MOG-specific proliferation compared with PBS-treated mice.
Conclusion: Electroporation of tolDC with MOG mRNA is an effective tool to generate MOG-presenting murine tolDC without affecting their tolerogenic properties. Moreover, MOG mRNA-electroporated tolDC effectively suppress EAE splenocytes in vitro and in vivo. These findings hold promise for further (pre)clinical translation. In this perspective, the clinical effect of MOG mRNA-electroporated tolDC treatment is currently being investigated in EAE mice.
Disclosure: This work was supported by a BOF-GOA Grant from the University of Antwerp, Belgium.
Judith Derdelinckx: holds a PhD Fellowship of the Research Foundation - Flanders (FWO), further nothing to disclose.
María José Mansilla: nothing to disclose.
Maxime De Laere: nothing to disclose.
Wai-Ping Lee: nothing to disclose.
Juan Navarro Barriuso: nothing to disclose.
Patrick Cras: nothing to disclose.
Barbara Willekens: nothing to disclose.
Peter Ponsaerts: nothing to disclose.
Zwi N. Berneman: nothing to disclose.
Eva María Martínez-Cáceres: nothing to disclose.
Nathalie Cools: nothing to disclose.