Autologous Tolerogenic Dendritic Cells Research Articles

Trial in Progress: An Open-Label, Multi-Center, First in Human, Phase 1/2a Trial to Evaluate the Safety and Preliminary Efficacy of Autologous Tolerogenic Dendritic Cells Ex Vivo Loaded with Recombinant Factor VIII (FVIII) in Adults with Congenital Hemophilia a (HA) with Neutralizing Antibodies to FVIII and Having Failed Immune Tolerance Induction (ITI)

Background Severe hemophilia A is a congenital bleeding disorder caused by a lack of factor VIII (FVIII) in plasma. Without treatment patients experience prolonged bleeding, either spontaneously or following trauma, and even life-threatening bleeds. Replacement therapy with FVIII is the standard of care, but development of neutralizing antibodies to FVIII is common, which reduces or completely prohibits the effect of FVIII treatment. Idogen AB has developed a cell therapy platform with the main purpose of making patients tolerant to FVIII and thus eliminating the problem of the immune system producing neutralizing antibodies. The cell therapy product, ItolDC-028, is produced from autologous mononuclear cells, collected by leukapheresis. A GMP-facility at Radboud University Hospital in the Netherlands enriches and cultures cells into tolerogenic dendritic cells, finally loaded with FVIII-antigen and cryopreserved. By this unique process the autologous cells has undergone a specific culture process to induce tolerance to FVIII. Trial Design and Methods This trial is a first-in-human, phase 1/2a, open-label trial, evaluating the safety and preliminary efficacy with ascending single doses of autologous tolerogenic dendritic cells for the management of neutralizing antibodies against FVIII in patients with hemophilia A. Male subjects aged 18-60 years and diagnosed with severe hemophilia A, with neutralizing antibodies against FVIII, having failed immune tolerance induction (ITI) treatment or for whom it is not possible to treat with ITI, a reported peak titer of ≥ 5 Bethesda Units (BU) and ≥ 1 BU at inclusion are eligible for the trial. Concomitant emicizumab is allowed, but FEIBA and other FVIII-containing treatments are excluded except in the case of an urgent need to treat bleeding. The primary objective is safety/tolerability of ItolDC-028, evaluated as the number of adverse events (AEs), changes in vital signs, physical examination findings and laboratory assessments including titers of neutralizing antibodies (BU) and ECG recordings from administration of ItolDC-028 up to the week 26 visit. The secondary objective is to evaluate the preliminary therapeutic efficacy of three dose levels of ItolDC-028 through evaluation of immune response, titer of neutralizing antibodies (BU), FVIII recovery and half-life as response to FVIII challenge at the week 20 visit. A minimum of nine subjects will be included in a classic 3+3 trial design, with single incremental dose levels of 36x106, 72x106 and 144x106 cells. Three additional subjects can be included in each dose level, upon reported signs of dose-limiting toxicity. An Internal Monitoring Board will evaluate the safety of the first subject at each dose level to decide if the next subject can be treated, and then all subjects at each dose level prior to escalation to the next dose level. There will be at least 2 weeks between treatments within a dose level and at least 6 weeks between the dose levels. Figure 1. At each clinical site, frozen ItolDC-028 is thawed and given to the subject by an intravenous infusion. Subjects are followed for 26 weeks including physical examinations, vital signs, laboratory assessments, FVIII-antibody titers, immunological changes, bleeding episodes and quality-of-life questionnaires (EQ-5D and Haem-A-QoL). In addition, there will be a long-term follow-up visit after 52 weeks. Figure 2. Twenty weeks after ItolDC-028 administration, the subject will be challenged by an administration of FVIII (Kovaltry®, 50 IU/kg), to evaluate the preliminary efficacy of ItolDC-028. The trial will be performed in eight European academic hospitals. The first subject is planned to be enrolled in August 2022. The trial is approved by the Swedish and Norwegian Competent Authorities and by the Swedish Ethical Review Authority (July 2022) and will also be reviewed by EU and UK Authorities. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

A Phase I/IIa study of autologous tolerogenic dendritic cells immunotherapy in kidney transplant recipients

Kidney transplant survival is shortened by chronic rejection and side effects of standard immunosuppressive drugs. Cell-based immunotherapy with tolerogenic dendritic cells has long been recognized as a promising approach to reduce general immunosuppression. Published trials report the safety and the absence of therapy-related adverse reactions in patients treated with tolerogenic dendritic cells suffering from several inflammatory diseases. Here, we present the first phase I clinical trial results using human autologous tolerogenic dendritic cells (ATDC) in kidney transplantation. Eight patients received ATDC the day before transplantation in conjunction with standard steroids, mycophenolate mofetil and tacrolimus immunosuppression with an option to taper mycophenolate mofetil. ATDC preparations were manufactured in a Good Manufacturing Practice-compliant facility and fulfilled cell count, viability, purity and identity criteria for release. A control group of nine patients received the same standard immunosuppression, except basiliximab induction replaced ATDC therapy and mycophenolate tapering was not allowed. During the three-year follow-up, no deaths occurred and there was 100% graft survival. No significant increase of adverse events was associated with ATDC infusion. Episodes of rejection were observed in two patients from the ATDC group and one patient from the control group. However, all rejections were successfully treated by glucocorticoids. Mycophenolate was successfully reduced/stopped in five patients from the ATDC group, allowing tacrolimus monotherapy for two of them. Regarding immune monitoring, reduced CD8 T cell activation markers and increased Foxp3 expression were observed in the ATDC group. Thus, our results demonstrate ATDC administration safety in kidney-transplant recipients.

Transfection of Vitamin D3-Induced Tolerogenic Dendritic Cells for the Silencing of Potential Tolerogenic Genes. Identification of CSF1R-CSF1 Signaling as a Glycolytic Regulator.

The use of autologous tolerogenic dendritic cells (tolDC) has become a promising strategy to re-establish immune tolerance in autoimmune diseases. Among the different strategies available, the use of vitamin D3 for the generation of tolDC (VitD3-tolDC) has been widely tested because of their immune regulatory properties. To identify molecules and pathways involved in the generation of VitD3-tolDC, we established an easy and fast gene silencing method based on the use of Viromer blue to introduce siRNA into monocytes on day 1 of culture differentiation. The analysis of the effect of CD209 (DC-SIGN) and CD115 (CSF1R) down-modulation on the phenotype and functionality of transfected VitD3-tolDC revealed a partial role of CD115 in their tolerogenicity. Further investigations showed that CSF1R-CSF1 signaling is involved in the induction of cell metabolic reprogramming, triggering glycolysis to produce high amounts of lactate, a novel suppressive mechanism of T cell proliferation, recently found in autologous tolerogenic dendritic cells (ATDCs).

Vitamin D3-Induced Tolerogenic Dendritic Cells Modulate the Transcriptomic Profile of T CD4+ Cells Towards a Functional Hyporesponsiveness.

The use of autologous tolerogenic dendritic cells (tolDC) has become a promising alternative for the treatment of autoimmune diseases. Among the different strategies available, the use of vitamin D3 for the generation of tolDC (vitD3-tolDC) constitutes one of the most robust approaches due to their immune regulatory properties, which are currently being tested in clinical trials. However, the mechanisms that vitD3-tolDC trigger for the induction of tolerance remain elusive. For this reason, we performed a full phenotypical, functional, and transcriptomic characterization of T cells upon their interaction with autologous, antigen-specific vitD3-tolDC. We observed a strong antigen-specific reduction of T cell proliferation, combined with a decrease in the relative prevalence of TH1 subpopulations and IFN-γ production. The analysis of the transcriptomic profile of T CD4+ cells evidenced a significant down-modulation of genes involved in cell cycle and cell response to mainly pro-inflammatory immune-related stimuli, highlighting the role of JUNB gene as a potential biomarker of these processes. Consequently, our results show the induction of a strong antigen-specific hyporesponsiveness combined with a reduction on the TH1 immune profile of T cells upon their interaction with vitD3-tolDC, which manifests the regulatory properties of these cells and, therefore, their therapeutic potential in the clinic.

Preclinical Assessment of Autologous Tolerogenic Dendritic Cells From End-stage Renal Disease Patients.

Kidney transplantation is the therapeutic of choice for patients with kidney failure. While immunosuppressive drugs can control graft rejection, their use is associated with increased infections and cancer, and they do not effectively control chronic graft rejection. Cell therapy is an attractive strategy to minimize the use of pharmacological drugs. We recently developed a protocol to generate human monocyte-derived autologous tolerogenic dendritic cells (ATDCs) from healthy volunteers. Herein, we transferred the ATDC manufacturing protocol to a Good Manufacturing Practice (GMP)-compliant facility. Furthermore, we compared the phenotype and in vitro functions of ATDCs generated from patients with end-stage renal disease to those generated from healthy volunteers. We describe the critical steps for GMP-compliant production of ATDCs and define the quality criteria required to allow release of the cell products. Furthermore, we showed that ATDCs generated from healthy volunteers and patients with kidney failure display the same tolerogenic profile based on their phenotype, resistance to maturation, and ability to modulate T-cell responses. Together, these results allowed us to define the production process and the quality criteria for the release of ATDCs before their administration in patients receiving a kidney transplant.

Human Tolerogenic Dendritic Cells Regulate Immune Responses through Lactate Synthesis

Cell therapy is a promising strategy for treating patients suffering from autoimmune or inflammatory diseases or receiving a transplant. Based on our preclinical studies, we have generated human autologous tolerogenic dendritic cells (ATDCs), which are being tested in a first-in-man clinical trial in kidney transplant recipients. Here, we report that ATDCs represent a unique subset of monocyte-derived cells based on phenotypic, transcriptomic, and metabolic analyses. ATDCs are characterized by their suppression of Tcell proliferation and their expansion of Tregs through secreted factors. ATDCs produce high levels of lactate that shape Tcell responses toward tolerance. Indeed, Tcells take up ATDC-secreted lactate, leading to a decrease of their glycolysis. Invivo, ATDCs promote elevated levels of circulating lactate and delay graft-versus-host disease by reducing Tcell proliferative capacity. The suppression of Tcell immunity through lactate production by ATDCs is a novel mechanism that distinguishes ATDCs from other cell-based immunotherapies.

Tolerogenic dendritic cell-based treatment for multiple sclerosis (MS): a harmonised study protocol for two phase I clinical trials comparing intradermal and intranodal cell administration

IntroductionBased on the advances in the treatment of multiple sclerosis (MS), currently available disease-modifying treatments (DMT) have positively influenced the disease course of MS. However, the efficacy of DMT is...

MAP7 and MUCL1 Are Biomarkers of Vitamin D3-Induced Tolerogenic Dendritic Cells in Multiple Sclerosis Patients.

The administration of autologous tolerogenic dendritic cells (tolDC) has become a promising alternative for the treatment of autoimmune diseases, such as multiple sclerosis (MS). Specifically, the use of vitamin D3 for the generation of tolDC (vitD3-tolDC) constitutes one of the most widely studied approaches, as it has evidenced significant immune regulatory properties, both in vitro and in vivo. In this article, we generated human vitD3-tolDC from monocytes from healthy donors and MS patients, characterized in both cases by a semi-mature phenotype, secretion of IL-10 and inhibition of allogeneic lymphocyte proliferation. Additionally, we studied their transcriptomic profile and selected a number of differentially expressed genes compared to control mature and immature dendritic cells for their analysis. Among them, qPCR results validated CYP24A1, MAP7 and MUCL1 genes as biomarkers of vitD3-tolDC in both healthy donors and MS patients. Furthermore, we constructed a network of protein interactions based on the literature, which manifested that MAP7 and MUCL1 genes are both closely connected between them and involved in immune-related functions. In conclusion, this study evidences that MAP7 and MUCL1 constitute robust and potentially functional biomarkers of the generation of vitD3-tolDC, opening the window for their use as quality controls in clinical trials for MS.

Potential Pharmacologic Targets for the Prevention of Rheumatoid Arthritis

Rheumatoid arthritis (RA) immunopathology starts many years before clinical disease manifests. An early event is the breakdown in B-cell tolerance and the emergence of autoantibodies. Some years later, the autoantibody response matures, with epitope spreading and isotype switching suggesting more focused T-cell involvement. Circulating proinflammatory cytokines and chemokines appear concurrently, marking a phase of preclinical inflammation. Eventually individuals develop musculoskeletal symptoms and fatigue, heralding the imminent onset of synovitis. The prevention of RA can be aligned to these disease stages, which may simplistically be viewed as breach of tolerance, maturation of autoimmunity, and subclinical inflammation. At each stage, an "ethical" balance must be struck between the potential for benefit versus harm, in large part determined by the likelihood of progression to RA. In particular, tolerogenic interventions should be favored during asymptomatic disease, providing long-lasting, possibly lifelong, benefit from a short-term intervention. During the breach-of-tolerance phase, tolerogenic interventions might involve the administration of autoantigenic peptides in an attempt to shut down autoreactive B cells. The peptides may be "naked," encapsulated in nanoparticles or loaded into autologous tolerogenic dendritic cells. It may also be possible to interfere with antigen generation and presentation, such as via peptidyl arginine deiminase inhibition. Attempts to interfere with B cells via depletion, differentiation, or intracellular signaling are also logical at this stage, if deemed sufficiently well tolerated. These interventions remain relevant during maturation of autoimmunity, but targeted T-cell interventions also become relevant, such as co-stimulation blockade and, potentially, interventions that target intracellular signaling pathways within T cells. Targeting of interleukin 23 during this phase may interfere with maturation of the autoantibody response by modulating autoantibody glycosylation, as well as by inhibiting T-helper 17 differentiation. During the phase of subclinical inflammation, targeting cytokines and chemokines could conceivably prevent progression to RA. Furthermore, tolerance induction is opposed by systemic inflammation and, during this phase, it may be necessary to consider therapeutic combinations of tolerogenic and antiinflammatory interventions.

Beyond the Magic Bullet: Current Progress of Therapeutic Vaccination in Multiple Sclerosis.

Multiple sclerosis (MS) is a chronic immune-mediated disease of the central nervous system (CNS) characterized by neuroinflammation, neurodegeneration and impaired repair mechanisms that lead to neurological disability. The crux of MS is the patient’s own immune cells attacking self-antigens in the CNS, namely the myelin sheath that protects nerve cells of the brain and spinal cord. Restoring antigen-specific tolerance via therapeutic vaccination is an innovative and exciting approach in MS therapy. Indeed, leveraging the body’s attempt to prevent autoimmunity, i.e., tolerization, focuses on the underlying cause of the disease and could be the key to solving neuroinflammation. In this perspective, antigen-specific vaccination targets only the detrimental and aberrant immune response against the specific disease-associated antigen(s) involved while retaining the capacity of the immune system to respond to unrelated antigens. We review the experimental approaches of tolerance-inducing vaccination in relapsing and progressive forms of MS that have reached the clinical development phase, including vaccination with autologous T cells, autologous tolerogenic dendritic cells, T cell receptor peptide vaccination, altered peptide ligand, ATX-MS-1467, cluster of differentiation (CD)-206-targeted liposomal myelin basic protein peptides and DNA vaccination. Failures, successes and future directions are discussed.

Towards a dendritic cell-based vaccine for the treatment of multiple sclerosis (MS): interim safety data of the first dose cohort of the MS-tolDC phase I clinical trial

Towards a dendritic cell-based vaccine for the treatment of multiple sclerosis (MS): interim safety data of the first dose cohort of the MS-tolDC phase I clinical trial

Tolerogenic dendritic cell therapy in organ transplantation.

Although the occurrence of acute rejection was significantly reduced and the allograft survival at 1 year was massively improved by the development of pharmacological immunosuppressive drugs, little progress has been made regarding long-term graft survival. Cell therapy appears to be an innovative and promising strategy to minimize the use of immunosuppression in transplantation and consequently increases long-term graft survival. The strength of cell therapy is that it will induce graft-specific tolerance and not a general immunosuppression of the patients. Several candidates, such as tolerogenic dendritic cells, have been gaining interest as an efficient means of promoting antigen-specific tolerance over recent years. Studies performed in rodent models have demonstrated the feasibility and efficacy of tolerogenic dendritic cells for the induction of tolerance in transplantation. In parallel, protocols to generate human tolerogenic dendritic cells in vitro have been defined, and some phase I clinical trials in autoimmune diseases have been recently performed to evaluate the safety of tolerogenic dendritic cell therapy. In this review, we will focus on the potential therapeutic interest of these cells in transplantation as well as their generation and characterization in humans. Finally, we will describe our current clinical trial using autologous tolerogenic dendritic cells in transplantation.

Autologous tolerogenic dendritic cells for rheumatoid and inflammatory arthritis

ObjectivesTo assess the safety of intra-articular (IA) autologous tolerogenic dendritic cells (tolDC) in patients with inflammatory arthritis and an inflamed knee; to assess the feasibility and acceptability of the approach...

DOP031. Phase I study of intraperitoneal administration of autologous tolerogenic dendritic cells for refractory Crohn's disease.

DOP031. Phase I study of intraperitoneal administration of autologous tolerogenic dendritic cells for refractory Crohn's disease.

Combining autologous dendritic cell therapy with CD3 antibodies promotes regulatory T cells and permanent islet allograft acceptance.

Cell therapy and the use of mAbs that interfere with T cell effector functions constitute promising approaches for the control of allograft rejection. In the current study, we investigated a novel approach combining administration of autologous tolerogenic dendritic cells with short-term treatment with CD3-specific Abs. Permanent acceptance of pancreatic islet allografts was achieved in mice treated with the combination therapy the day before transplantation but not in recipients treated with either therapy alone. The combination treatment induced a marked decrease in T cells infiltrating the allografts and a sustained reduction of antidonor responses. Importantly, CD4(+)Foxp3(+) regulatory T cells appeared to play a crucial role in the long-term graft acceptance. Their frequency increased significantly in the spleen, draining lymph nodes, and transplanted islets and remained elevated over the long term; they exhibited increased donor-specific suppressive functions; and their removal at the time of transplantation abrogated the therapeutic effect of the combined therapy. These results support the therapeutic potential of protocols combining autologous dendritic cells and low-dose CD3 Abs, both currently in clinical development, and that act in synergy to control allogeneic immune responses and favor graft survival in a full-mismatch situation.

A1.43 Autologous tolerogenic dendritic cells in rheumatoid and inflammatory arthritis

Background and ObjectivesCurrent therapies used in the treatment of rheumatoid and inflammatory arthritis, although efficacious in treating symptoms, may non-specifically suppress the immune system with consequences such as increased risk...

Immunotherapy with Tolerogenic Dendritic Cells Alone or in Combination with Rapamycin Does Not Reverse Diabetes in NOD Mice

Type 1 diabetes is a metabolic disease caused by autoimmunity towards β-cells. Different strategies have been developed to restore β-cell function and to reestablish immune tolerance to prevent and cure the disease. Currently, there is no effective treatment strategy to restore endogenous insulin secretion in patients with type 1 diabetes. This study aims to restore insulin secretion in diabetic mice with experimental antigen-specific immunotherapy alone or in combination with rapamycin, a compound well known for its immunomodulatory effect. Nonobese diabetic (NOD) mice develop spontaneous type 1 diabetes after 12 weeks of age. Autologous tolerogenic dendritic cells—consisting in dendritic cells pulsed with islet apoptotic cells—were administered to diabetic NOD mice alone or in combination with rapamycin. The ability of this therapy to revert type 1 diabetes was determined by assessing the insulitis score and by measuring both blood glucose levels and C-peptide concentration. Our findings indicate that tolerogenic dendritic cells alone or in combination with rapamycin do not ameliorate diabetes in NOD mice. These results suggest that alternative strategies may be considered for the cure of type 1 diabetes.

Adoptive Infusion of Tolerogenic Dendritic Cells Prolongs the Survival of Pancreatic Islet Allografts: A Systematic Review of 13 Mouse and Rat Studies

ObjectiveThe first Phase I study of autologous tolerogenic dendritic cells (Tol-DCs) in Type 1 diabetes (T1D) patients was recently completed. Pancreatic islet transplantation is an effective therapy for T1D, and infusion of Tol-DCs can control diabetes development while promoting graft survival. In this study, we aim to systematically review islet allograft survival following infusion of Tol-DCs induced by different methods, to better understand the mechanisms that mediate this process.MethodsWe searched PubMed and Embase (from inception to February 29th, 2012) for relevant publications. Data were extracted and quality was assessed by two independent reviewers. We semiquantitatively analyzed the effects of Tol-DCs on islet allograft survival using mixed leukocyte reaction, Th1/Th2 differentiation, Treg induction, and cytotoxic T lymphocyte activity as mechanisms related-outcomes. We discussed the results with respect to possible mechanisms that promote survival.ResultsThirteen articles were included. The effects of Tol-DCs induced by five methods on allograft survival were different. Survival by each method was prolonged as follows: allopeptide-pulsed Tol-DCs (42.14±44 days), drug intervention (39 days), mesenchymal stem cell induction (23 days), genetic modification (8.99±4.75 days), and other derivation (2.61±6.98 days). The results indicate that Tol-DC dose and injection influenced graft survival. Single-dose injections of 104 Tol-DCs were the most effective for allograft survival, and multiple injections were not superior. Tol-DCs were also synergistic with immunosuppressive drugs or costimulation inhibitors. Possible mechanisms include donor specific T cell hyporesponsiveness, Th2 differentiation, Treg induction, cytotoxicity against allograft reduction, and chimerism induction.ConclusionsTol-DCs induced by five methods prolong MHC mismatched islet allograft survival to different degrees, but allopeptide-pulsed host DCs perform the best. Immunosuppressive or costimulatory blockade are synergistic with Tol-DC on graft survival. Multiple injections are not superior to single injection. Yet more rigorously designed studies with larger sample sizes are still needed in future.

A Critical Role for Graft Passenger Leukocytes in Donor-Specific Allograft Tolerance Induced by Autologous Tolerogenic Dendritic Cells

A Critical Role for Graft Passenger Leukocytes in Donor-Specific Allograft Tolerance Induced by Autologous Tolerogenic Dendritic Cells

Tolerogenic dendritic cells and negative vaccination in transplantation: from rodents to clinical trials

The use of immunosuppressive (IS) drugs to treat transplant recipients has markedly reduced the incidence of acute rejection and early graft loss. However, such treatments have numerous adverse side effects and fail to prevent chronic allograft dysfunction. In this context, therapies based on the adoptive transfer of regulatory cells are promising strategies to induce indefinite transplant survival. The use of tolerogenic dendritic cells (DC) has shown great potential, as preliminary experiments in rodents have demonstrated that administration of tolerogenic DC prolongs graft survival. Recipient DC, Donor DC, or Donor Ag-pulsed recipient DC have been used in preclinical studies and administration of these cells with suboptimal immunosuppression increases their tolerogenic potential. We have demonstrated that autologous unpulsed tolerogenic DC injected in the presence of suboptimal immunosuppression are able to induce Ag-specific allograft tolerance. We derived similar tolerogenic DC in different animal models (mice and non-human primates) and confirmed their protective abilities in vitro and in vivo. The mechanisms involved in the tolerance induced by autologous tolerogenic DC were also investigated. With the aim of using autologous DC in kidney transplant patients, we have developed and characterized tolerogenic monocyte-derived DC in humans. In this review, we will discuss the preclinical studies and describe our recent results from the generation and characterization of tolerogenic monocyte-derived DC in humans for a clinical application. We will also discuss the limits and difficulties in translating preclinical experiments to theclinic.