Generation and characterisation of therapeutic tolerogenic dendritic cells for rheumatoid arthritis

Abstract

ObjectivesTolerogenic dendritic cells (tolDCs) constitute a promising experimental treatment for targeting autoreactive T cells in autoimmune diseases, including rheumatoid arthritis (RA). The authors' goal is to bring tolDC therapy for RA to the clinic. Here the authors address key translational issues related to the manufacturing of tolDCs from RA patients with current good manufacturing practice (cGMP)-compliant reagents, the stability of tolDCs, and the selection of suitable quality control markers.MethodsHuman monocyte-derived tolDCs were established from RA patients and healthy controls (HCs) using the immunosuppressive drugs dexamethasone and vitamin D3, and the cGMP-grade immunomodulator, monophosphoryl lipid A, in the cGMP-compliant medium, CellGroDC. The functionality of tolDCs and tolDC-modulated autologous CD4 T cells was determined by flow cytometry, [3H]thymidine incorporation and ELISA.ResultsClinical-grade tolDCs established from patients with RA exhibit a typical tolerogenic phenotype of reduced costimulatory molecules, low production of proinflammatory cytokines and impaired stimulation of autologous antigen-specific T cells, comparable to HC tolDCs. Toll-like receptor 2 (TLR-2) was highly expressed by tolDCs but not mature DCs. Furthermore, tolDCs suppressed mature DC-induced T cell proliferation, interferon γ and interleukin 17 production, and rendered T cells hyporesponsive to further stimulation. Importantly, tolDCs were phenotypically stable in the absence of immunosuppressive drugs and were refractory to further challenge with proinflammatory mediators.ConclusionstolDCs established from patients with RA are comparable to those derived from healthy donors. TLR-2 was identified as an ideal marker for quality control of tolDCs. Potently tolerogenic and highly stable, these tolDCs are a promising cellular therapeutic for tailored immunomodulation in the treatment of RA.

Lipopolysaccharide‐primed heterotolerant dendritic cells suppress experimental autoimmune uveoretinitis by multiple mechanisms

“…Similar results have been found in other models and also in parallel clinical studies using autologous DC 69, 70…”

Stable antigen‐specific T‐cell hyporesponsiveness induced by tolerogenic dendritic cells from multiple sclerosis patients

“…Use of tolDCs to induce antigen-specific anergy and/or Treg cells has been tested in several autoimmune animal models [18], including the EAE model [19]. Hence, several groups have designed immunotherapeutic approaches involving administration of tolDCs to humans to induce T-cell tolerance in transplantation and autoimmune diseases [20,21]. Different strategies have been evaluated to generate tolDCs in vitro, as recently reviewed [18,22].…”

Immunomodulation of ALT-2 and TLR may collude in antigen specific T cell hyporesponsiveness: Proposed mechanism for elevated IL-10 levels in Balb/C mice

“…However the adjuvant formulation immunized, failed to induce proliferation and recall-response to the ALT-2 antigen. This could be due to (1) low expression of co-stimulatory signals on APC (2) low production of proinflammatory cytokines (3) in sufficient production of memory cells, which result in impaired T cell proliferation, rendering T cells tolerant (Harry et al 2010).…”