Generating myelin-expressing tolerogenic dendritic cells using mRNA electroporation: potential players for a cellular vaccine for the treatment of multiple sclerosis?
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1
University of Antwerp, Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, Belgium
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2
Antwerp University Hospital, Division of Neurology, Belgium
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3
Antwerp University Hospital, Center for Cell Therapy and Regenerative Medicine, Belgium
BACKGROUND: In the pursuit of re-establishing tolerance in autoimmune disorders such as multiple sclerosis (MS), the use of antigen-loaded tolerogenic dendritic cells (tolDC) is a promising strategy to restore immune balance in a disease antigen-specific manner. For antigen loading of tolDC, electroporation with mRNA encoding full-length proteins might offer the potential to prevent epitope spreading by inducing presentation of all myelin epitopes in a non-HLA-restricted manner.
OBJECTIVES: To generate antigen-expressing human tolDC using mRNA electroporation.
METHODS: In this study, 1-alpha,25-dihydroxyvitamin D3-treated monocyte-derived tolDC were electroporated with mRNA encoding enhanced green fluorescent protein (eGFP) and myelin oligodendrocyte glycoprotein (MOG). To validate the electroporation procedure, mRNA and protein levels were analyzed in function of time following electroporation using qRT-PCR and flow cytometry or Western blot.
RESULTS: Flow cytometric analysis of eGFP expression revealed a transfection efficiency of 68.6% (SD ±3,4%) for immature tolDC (itolDC) and 54.4% (SD ±29.7%) for cytokine-activated tolDC, demonstrating the feasibility to electroporate tolDC. MOG protein expression was detectable until at least 72 hours after mRNA electroporation as evidenced by Western blot analysis. Importantly, no difference in the expression of the costimulatory markers CD80 and CD86 by electroporated (i)tolDC nor in their capacity to induce T cell hyporesponsiveness in an allogeneic mixed lymphocyte reaction was found following mRNA electroporation when compared to mock electroporated and/or unelectroporated (i)tolDC.
CONCLUSION: Our results demonstrate that mRNA electroporation of tolDC effectively induces MOG expression without affecting their tolerogenic properties. The capacity of mRNA-electroporated tolDC to present antigen in a specific manner is currently being assessed by evaluating their modulatory effect on myelin-specific T cell responses. Ultimately, therapeutic vaccination with tolDC electroporated with mRNA encoding full-length myelin-derived proteins may lead to a more effective therapy for MS by induction of T cell tolerance in a myelin-specific manner.
Samenvatting in het Nederlands:
Naarmate de basismechanismen achter autoimmuunziekten zoals multiple sclerose (MS) verder ontrafeld worden, vormen celgebaseerde therapieën een veelbelovende strategie voor de behandeling van deze aandoeningen. Dendritische cellen (DC) zijn gespecialiseerde witte bloedcellen die afweerreacties kunnen aansturen dan wel onderdrukken. De afweeronderdrukkende of zogenaamde tolerogene DC vormen een interessante celgroep in het onderzoek naar celbehandelingen voor MS.
In deze studie onderzochten we een specifieke techniek voor het opladen van tolerogene DC met de eiwitten waartegen de afweerreactie bij MS gericht is. Op die manier trachten we een celgebaseerde behandeling te ontwikkelen die de immuunrespons bij MS-patiënten op een gerichte manier kan onderdrukken.
Résumé en Français:
Le progrès des connaissances sur les causes de maladies auto-immunes tels que la sclérose en plaques (SEP) permet de déveloper de nouvelles stratégies thérpeutiques prometteuses basées sur la thérapie cellulaire. Les cellules dendritiques (CD) sont des globules blancs specialisés qui permettent de susciter ou de réprimer des reactions de défense. Les CD dites “tolérogènes” qui répriment les mécanismes de défense forment une population de cellules intéressante dans la thérapie cellulaire de la SEP. Dans cette etude nous avons étudié une technique spécifique qui permet de charger les CD tolérogènes avec des protéines contre lesquelles est dirigée la reaction autoimmune dans la SEP. De cette façon nous essayons de mettre au point une thérapie cellulaire qui permettra d’inhiber de manière ciblée la réponse immune chez des patients atteints de SEP.
Keywords:
Dendritic Cells,
MRNA electroporation,
Multiple Sclerosis,
antigen-specific tolerance induction,
cellular therapy
Conference:
6th Belgian Brain Congress, MONS, Belgium, 8 Oct - 8 Oct, 2016.
Presentation Type:
Oral Presentation
Topic:
Brain and brain diseases: between heredity and environment
Citation:
Derdelinckx
J,
Lee
W,
De Laere
M,
Cras
P,
Willekens
B,
Berneman
ZN and
Cools
N
(2016). Generating myelin-expressing tolerogenic dendritic cells using mRNA electroporation: potential players for a cellular vaccine for the treatment of multiple sclerosis?.
Conference Abstract:
6th Belgian Brain Congress.
doi: 10.3389/conf.fnagi.2016.03.00022
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Received:
29 Jun 2016;
Published Online:
01 Jul 2016.
*
Correspondence:
Dr. Judith Derdelinckx, University of Antwerp, Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, Edegem, 2650, Belgium, judith.derdelinckx@student.uantwerpen.be