Immune tolerance in multiple sclerosis and neuromyelitis optica with peptide-loaded tolerogenic dendritic cells in a phase 1b trial.

Zubizarreta I, Flórez-Grau G, Vila G, Cabezón R, España C, Andorra M, Saiz A, Llufriu S, Sepulveda M, Sola-Valls N, Martinez-Lapiscina EH, Pulido-Valdeolivas I, Casanova B, Martinez Gines M, Tellez N, Oreja-Guevara C, Español M, Trias E, Cid J, Juan M, Lozano M, Blanco Y, Steinman L, Benitez-Ribas D, Villoslada P
Source: Proc Natl Acad Sci USA
Publication Date: (2019)
Issue: 116(17): 8463-8470
Research Area:
Immunotherapy / Hematology
Culture Media:

In the publication of the authors describe a phase 1b clinical trial to investigate the safety and feasibility of tolDC therapy for the CNS-related autoimmune disorders Multiple Sclerosis and Neuromyelitis optica with peptide-loaded tolerogenic dendritic cells.

Autologous tolerogenic dendritic cells have been generated from patients PBMCS under GMP compliant conditions using the Lonza X-VIVO Medium.
Monocytes have cultured in In the protocol  in the presence of 500 IU (international units)/mL of IL-4 and 800 IU/mL of GM-CSF during 7 d in X-VIVO-15 (BioWhittaker; Lonza) supplemented with 2% autologous serum.

At day 3, fresh medium and cytokines were added; furthermore, 10-6 M dexamethasone was added to the cells to induce the tolerogenic phenotype.

At day 6, to boost peptide-specific DC tolerogenic properties, cytokines including IL-1ß, IL-6, TNF-a, and prostaglandin E2, as well as the immunogenic peptides , were added for 24 h and washed before administration. TolDC from MS patients were loaded with seven myelin peptides and DC from NMOSD patients were stimulated with peptides from AQP4 patients.

Patients received intravenous administration of TolDC every two weeks (week 0 , 2 and 4 ) representing a total of three administrations per patient. The dose escalation  (50 × 10exp6, 100 × 10exp6, 150 × 10exp6, and 300 × 10exp6 DCs) occured as expected in the absence of limiting toxicity in the previous dosage level. 


There are adaptive T-cell and antibody autoimmune responses to myelin-derived peptides in multiple sclerosis (MS) and to aquaporin-4 (AQP4) in neuromyelitis optica spectrum disorders (NMOSDs). Strategies aimed at antigen-specific tolerance to these autoantigens are thus indicated for these diseases. One approach involves induction of tolerance with engineered dendritic cells (tolDCs) loaded with specific antigens. We conducted an in-human phase 1b clinical trial testing increasing concentrations of autologous tolDCs loaded with peptides from various myelin proteins and from AQP4. We tested this approach in 12 patients, 8 with MS and 4 with NMOSD. The primary end point was the safety and tolerability, while secondary end points were clinical outcomes (relapses and disability), imaging (MRI and optical coherence tomography), and immunological responses. Therapy with tolDCs was well tolerated, without serious adverse events and with no therapy-related reactions. Patients remained stable clinically in terms of relapse, disability, and in various measurements using imaging. We observed a significant increase in the production of IL-10 levels in PBMCs stimulated with the peptides as well as an increase in the frequency of a regulatory T cell, known as Tr1, by week 12 of follow-up. In this phase 1b trial, we concluded that the i.v. administration of peptide-loaded dendritic cells is safe and feasible. Elicitation of specific IL-10 production by peptide-specific T cells in MS and NMOSD patients indicates that a key element in antigen specific tolerance is activated with this approach. The results warrant further clinical testing in larger trials.