Phase I study of tumor Ag-loaded IL-12 secreting semi-mature DC for the treatment of pediatric cancer

doi:10.1080/14653240701589221

Cytotherapy

Volume 9, Issue 8, 2007, Pages 755-770

https://doi.org/10.1080/14653240701589221 Get rights and content

Background

Cancer vaccines employing DC in their capacity as APC have been tolerated well and have shown some efficacy in clinical studies. IL-12, a cytokine critical for type 1 T-helper (Th1) lymphocyte and cytotoxic T-lymphocyte (CTL) differentiation, when released from a DC-based cancer vaccine, may support the generation of a cellular T-cell response.

Methods

We applied tumor cell lysate plus keyhole limpet hemocyanin (KLH)-loaded and 48-h lipopolysaccharide (LPS) plus IFN-γ-stimulated fully mature DC, which do not release IL-12, subcutaneously to eight patients, and maximally 6-h stimulated semi-mature (sm) DC, which are potent producers of IL-12, subcutaneously (n=6) or intranodally (n=8) as a cancer vaccine to patients suffering from advanced solid pediatric malignancies.

Results

No serious adverse events were observed following application of IL-12-releasing smDC. Following immunization the majority of patients responded positively to KLH in a delayed-type hypersensitivity (DTH) test. In addition, three of six intranodally treated patients responded to the tumor Ag in the DTH test.

Discussion

We conclude that treatment with a DC-based cancer vaccine enabled to release the immune regulatory cytokine IL-12 is safe and feasible and has the potential to induce a cellular immune response in pediatric cancer patients.

Introduction

Although still at an early stage of clinical development, the concept of applying rationally designed cancer vaccines may have the potential to change cancer treatment. DC as the principal regulators of immunity [1] have received special attention as a novel form of adjuvant for cancer immune therapy. Clinical trials using DC-based cancer vaccines have been designed for the treatment of various neoplastic diseases [2], and DC-based cancer vaccination for solid pediatric tumors has been reported from three centers [3, 4]. In most of these trials, some in vivo and/or in vitro evidence for the generation of anti-tumor immunity was found. The reported side-effects were usually mild and did not limit the application of the cancer vaccine. The majority of these studies were pilot or phase I trials designed to evaluate feasibility and toxicity but not the efficacy of DC-based cancer vaccination. Anecdotal reports of clinical benefits, encouraging as they may be, must be interpreted with caution at this stage of research.

An important issue in the design of a DC-based cancer vaccine concerns the decision of whether to use immature or mature DC. The use of mature DC, as suggested by some investigators [5], has the advantage that these DC are less likely to induce tolerance. In an immature state, DC act as sentinels in peripheral tissues, constantly sampling their environment by taking up and processing Ag. Bacterial products, by engagement of pattern recognition receptors 6., 7., 8., 9., virus-derived signals mediated by type I IFN 10., 11., 12. or T-lymphocyte-derived licensing signals mediated by the interaction of CD40 with its ligand CD154 13., 14., 15., 16., trigger a maturation process. Maturation causes DC to shut down their phagocytosis machinery and up-regulate the expression of MHC molecules and T-lymphocyte co-stimulatory receptors such as CD80 and CD86, while migrating into draining lymph nodes. This pattern recognition by immature DC determines the nature of the maturation process, resulting in distinct DC phenotypes with specific immune polarizing features [17]. DC characterized by release of IL-12 (type 1 DC; DC1) prime helper T lymphocytes (Th) to differentiate into IFN-γ-releasing type 1 helper (Th1) cells. In contrast, DC that do not release IL-12 (type 2 DC; DC2) guide the differentiation of IL-4-secreting Th2 cells [18, 19]. A Th1-mediated immune response supports cytolytic T lymphocytes (CTL) whereas Th2 cells support the production of Ab from B lymphocytes [20].

Collectively, these data suggest that a type 1 immune response guided by IL-12-secreting DC1 would be particularly effective in inducing cytolytic anti-tumor immunity. We have therefore developed a DC culture system designed to maximize IL-12 secretion by maturation with lipopolysaccharide (LPS), a potent bacterial endotoxin, and IFN-γ [21, 22]. Because IL-12 secretion from DC1 is limited to the first 24 h after maturation [23], we applied DC after a maximal 6 h of cultivation in the presence of LPS and IFN-γ as semi-mature actively IL-12-secreting type 1 DC (smDC1).

This study is the first to explore the utility of type 1 polarizing IL-12-secreting smDC1 in a clinical situation, taking into consideration well-understood immunologic concepts regarding the induction of cytolytic immunity in general and anti-tumor immunity in particular. The patients included in this clinical pilot trial suffered from advanced solid childhood malignancies. We were able to document the feasibility and lack of adverse events in the patients treated in this trial. Our data provide the basis for a larger randomized clinical study that is designed to document the efficacy and clinical benefit of cancer vaccination with smDC1.

Section snippets

Patient characteristics

Twenty pediatric and adolescent patients suffering from childhood solid extra-cranial tumors were included in this trial. Two patients were listed twice (JS012/JS056 and LS020/LS029) because they received one series of subcutaneous and one series of intranodal DC application. Thirteen patients had exhausted standard multimodal treatment options and were treated in a progressive disease (PD) status, one patient was in PR, two patients were in first CR, five patients were in second CR and one

Manufacturing and characterization of the cancer vaccine

Patient monocytes were differentiated into immature DC during a 5-day culture in the presence of GM-CSF and IL-4. These immature DC were loaded with autologous tumor cell lysate and the tracer Ag KLH. Subsequently, the charged immature DC were exposed to LPS plus IFN-γ to trigger maturation into IL-12-secreting smDC1. Quality control analysis was performed on one aliquot of smDC1 that was recovered from freezing and recultivated to allow differentiation into fully matured DC (fmDC). The quality

Discussion

Vaccination represents a true success story in modern medicine, and its principles, at least in the case of infectious diseases, have been found to be valid in different species. The field of tumor immunology has expanded considerably following the identification of DC as the major specialized APC of the immune system [1]. As DC go through distinct maturation stages during development, they are characterized by typical changes in their immune phenotype and their functional capabilities. Human

Acknowledgements

This work was supported by numerous private donations to the Children's Cancer Research Institute, Vienna, Austria, that partly funded this study, and by grants from the Austrian Industrial Research Promotion Fund (Forschungsförderungsfond der Gewerblichen Wirtschaft, Projektnummer 804445) and from the Vienna Business Agency (Wiener Wirtschaftsförderungsfond).

For including patients in our trial, we wish to thank Drs Ernst Horcher, Department for Pediatric Surgery, Medical University of Vienna,

References (50)

KalinskiP. et al.
T-cell priming by type-1 and type-2 polarized dendritic cells: the concept of a third signal
Immunol Today
(1999)
FelzmannT. et al.
Monocyte enrichment from leukapharesis products for the generation of DCs by plastic adherence, or by positive or negative selection
Cytotherapy
(2003)
BrossartP. et al.
Induction of cytotoxic T-lymphocyte responses in vivo after vaccinations with peptide-pulsed dendritic cells
Blood
(2000)
GillietM. et al.
Intranodal injection of semimature monocyte-derived dendritic cells induces T helper type 1 responses to protein neoantigen
Blood
(2003)
BanchereauJ. et al.
Immunobiology of dendritic cells
Annu Rev Immunol
(2000)
FelzmannT. et al.
Dendritic cells as adjuvants in antitumor immune therapy
Onkologie
(2002)
GeigerJ.D. et al.
Vaccination of pediatric solid tumor patients with tumor lysate-pulsed dendritic cells can expand specific T cells and mediate tumor regression
Cancer Res
(2001)
DagherR. et al.
Pilot trial of tumor-specific peptide vaccination and continuous infusion interleukin-2 in patients with recurrent Ewing sarcoma and alveolar rhabdomyosarcoma: an inter-institute NIH study
Med Pediatr Oncol
(2002)
LabeurM.S. et al.
Generation of tumor immunity by bone marrow-derived dendritic cells correlates with dendritic cell maturation stage
J Immunol
(1999)
MazzaccaroR.J. et al.
Major histocompatibility class I presentation of soluble antigen facilitated by Mycobacterium tuberculosis infection
Proc Natl Acad Sci USA
(1996)

SimmonsC.P. et al.

MHC class I-restricted cytotoxic lymphocyte responses induced by enterotoxin-based mucosal adjuvants

J Immunol

(1999)

ChoH.J. et al.

Immunostimulatory DNA-based vaccines induce cytotoxic lymphocyte activity by a T-helper cell-independent mechanism

Nat Biotechnol

(2000)

HamiltonS.E. et al.

Listeria monocytogenes infection overcomes the requirement for CD40 ligand in exogenous antigen presentation to CD8(+) T cells

J Immunol

(2001)

SigalL.J. et al.

Cytotoxic T-cell immunity to virus-infected non-haematopoietic cells requires presentation of exogenous antigen

Nature

(1999)

MuellerS.N. et al.

Rapid cytotoxic T lymphocyte activation occurs in the draining lymph nodes after cutaneous herpes simplex virus infection as a result of early antigen presentation and not the presence of virus

J Exp Med

(2002)

Le BonA. et al.

Cross-priming of CD8⁺ T cells stimulated by virus-induced type I interferon

Nat Immunol

(2003)

RidgeJ.P. et al.

A conditioned dendritic cell can be a temporal bridge between a CD4⁺ T-helper and a T-killer cell

Nature

(1998)

BennettS.R. et al.

Help for cytotoxic-T-cell responses is mediated by CD40 signalling

Nature

(1998)

SchoenbergerS.P. et al.

T-cell help for cytotoxic T lymphocytes is mediated by CD40–CD40L interactions

Nature

(1998)

LefrancoisL. et al.

Soluble antigen and CD40 triggering are sufficient to induce primary and memory cytotoxic T cells

J Immunol

(2000)

LiuY-J. et al.

Dendritic cell lineage, plasticity and cross-regulation

Nat Immunol

(2001)

TrinchieriG.

Interleukin-12 and the regulation of innate resistance and adaptive immunity

Nat Rev Immunol

(2003)

MosmannT.R. et al.

TH1 and TH2 cells: different patterns of lymphokine secretion lead to different functional properties

Annu Rev Immunol

(1989)

FelzmannT. et al.

Semi-mature IL-12 secreting dendritic cells present exogenous antigen to trigger cytolytic immune responses

Cancer Immunol Immunother

(2005)

HüttnerK.G. et al.

Generation of potent anti-tumor immunity in mice by interleukin-12-secreting dendritic cells

Cancer Immunol Immunother

(2005)

Cited by (68)

The generation and application of antigen-specific T cell therapies for cancer and viral-associated disease
2022, Molecular Therapy
Immunotherapy with antigen-specific T cells is a promising, targeted therapeutic option for patients with cancer as well as for immunocompromised patients with virus infections. In this review, we characterize and compare current manufacturing protocols for the generation of T cells specific to viral and non-viral tumor-associated antigens. Specifically, we discuss: (1) the different methodologies to expand virus-specific T cell and non-viral tumor-associated antigen-specific T cell products, (2) an overview of the immunological principles involved when developing such manufacturing protocols, and (3) proposed standardized methodologies for the generation of polyclonal, polyfunctional antigen-specific T cells irrespective of donor source. Ex vivo expanded cells have been safely administered to treat numerous patients with virus-associated malignancies, hematologic malignancies, and solid tumors. Hence, we have performed a comprehensive review of the clinical trial results evaluating the safety, feasibility, and efficacy of these products in the clinic. In summary, this review seeks to provide new insights regarding antigen-specific T cell technology to benefit a rapidly expanding T cell therapy field.
Systematic review of the immunological landscape of Wilms tumors
2021, Molecular Therapy Oncolytics
Citation Excerpt :
With respect to vaccines, we identified only one approach done for advanced solid pediatric cancer in which WT patients were included. The vaccine was based on an IL-12-secreting dendritic cell (DC)-based vaccine using tumor lysates.36 In one of the WT patients, the vaccine was associated with the transition of a progressive disease to stable disease 6 months after vaccine application.
Results of immunotherapy in childhood solid cancer have been so far, with the exception of neuroblastoma, quite disappointing. Lack of knowledge of the immune contexture of these tumors may have contributed to the failure of immunotherapies so far. Here, we systematically reviewed the literature regarding the immunology of Wilms tumor (WT), one of the most frequent pediatric solid tumors of the abdomen. In Wilms tumor patients the high cure rate of >90%, achieved by the combination of surgery and radio-chemotherapy, is at the expense of a high early and late toxicity. Moreover, treatment-resistant entities, such as diffuse anaplastic tumors or recurrent disease, still pose unsolved clinical problems. Successful immunotherapy could represent a novel and possibly less-toxic treatment option. Employing the PRISMA (Preferred Reporting Items for Systematic Review and Meta-Analysis) method of literature search, we analyzed the current knowledge of the immunological landscape of Wilms tumors in terms of tumor microenvironment, prognostic implications of single biomarkers, and immunotherapy response.
The efficacy of dendritic cell vaccine for newly diagnosed glioblastoma: A meta-analysis of randomized controlled studies
2021, Neurochirurgie
Citation Excerpt :
Dentritic cells showed a potent immune stimulatory mode of action after the stimulus of pathogen-associated microbial pattern molecules [28,29]. Interleukin (IL)-12-secreting dentritic cells could trigger robust helper T-lymphocyte type 1 and cytotoxic T-lymphocyte dominated immune responses [30–33]. The immunostimulatory capability of dentritic cells may show some ability to treat cancers because of immunosuppressive features [32,34,35].
The efficacy of dendritic cell vaccine to treat glioblastoma remained elusive and therefore we conducted a meta-analysis to explore the influence of dendritic cell vaccine on treatment efficacy of glioblastoma.
PubMed, EMbase, Web of science, EBSCO and Cochrane library databases have been searched through October 2020, and we included randomized controlled trials (RCTs) assessing the efficacy of dendritic cell vaccine for glioblastoma.
Four RCTs and 267 patients were included in the meta-analysis. Compared to control group for glioblastoma, dendritic cell vaccine demonstrated no obvious impact on overall survival (HR = 0.59; 95% CI = 0.34 to 1.04; P = 0.07), progression-free survival (PFS, HR = 0.72; 95% CI = 0.52 to 1.00; P = 0.05), nervous system disorders (OR = 0.61; 95% CI = 0.29 to 1.29; P = 0.20), or adverse events (OR = 1.44; 95% CI = 0.82 to 2.50; P = 0.20).
Dendritic cell vaccine may be not effective to treat glioblastoma.
The immune environment of bone sarcomas
2021, Bone Cancer: Bone Sarcomas and Bone Metastases - From Bench to Bedside
Despite advances in our understanding of cancer biology, the development of standardized, effective local control measures with surgery and radiation, and the emergence of targeted therapies, bone sarcoma survival rates remain unchanged for more than 30 years. Bone sarcomas still represent therapeutic challenges, and the low success of targeted therapies in these tumors highlights the fact that we have to take their environment particularly the immune part of it into account.
In the past 40 years, there has been remarkable progress in both the understanding of the immune system and of the ways to turn it on and off to treat cancer. Immunotherapy strategies for sarcomas are comparatively less advanced. Evidence from mouse models supports a major role for the immune system in preventing the development and progression of these cancers. In terms of immunology, bone sarcomas were long considered as cold tumors or event as immune desert for the cartilaginous tumors like chondrosarcoma and chordoma. Over the recent years, studies have been conducted to describe these tumors' immune environment. The goal is to better understand the interrelations between tumor and the immune components in order to design therapies adapted to bone sarcoma immune specificities. The purpose of this chapter is to describe the knowledge of the immune environment of three major bone tumors (osteosarcoma, chondrosarcoma, and chordoma) and the immunotherapies that are in development and could offer therapeutic options for these tumors.
Bispecific antibody activated T cells: A newly developed T cells with enhanced proliferation ability and cytotoxicity
2020, Immunology Letters
Adoptive cell therapy using ex vivo expanded lymphocytes has shown remarkable efficacy in tumor immunotherapy recently. Among various transfused immune cells, T lymphocytes are the most widely used since they are critical mediators of the immune system and have the capacity to kill tumor cells. However, there are drawbacks in the expanded T cells for transfusion including limited cytotoxicity, limited proliferation and lack of specificity. To improve the quality of these ex vivo expanded T cells, we have designed a new method to expand a group of T cells which are named bispecific antibodies activated T cells. It is the first time that such cells are induced by introducing the bispecific antibody drug (blinatumomab) and feeder cells (normal B cells and irradiated B cell originated lymphoma cells) to the traditional T cells culture system. Culture of freshly isolated human peripheral blood mononuclear cells in this newly designed cell culture system enabled these expanded T cells that (a) displayed a robust proliferation ability; (b) showed fully activated phenotype and enhanced cytokines production; (c) had a low proportion of CD4⁺CD25⁺ T regulatory cells and (d) exhibited strengthened cytotoxicity at relatively low effector: target ratios. This work further confirmed the feasibility of rapid induction and expansion of large amounts of human T cells in vitro by using bispecific antibodies and feeder cells. This strategy could also be used for other immune cells rapid expansion and help to improve the quality of these expanded immune cells for adoptive transfusion.
Spheroid glioblastoma culture conditions as antigen source for dendritic cell-based immunotherapy: spheroid proteins are survival-relevant targets but can impair immunogenic interferon γ production
2019, Cytotherapy
Glioblastoma is the most aggressive type of brain cancer. Dendritic cell (DC)-based immunotherapy against glioblastoma depends on the effectiveness of loaded antigens. Sphere-inducing culture conditions are being studied by many as a potential antigen source. Here, we investigated two different in vitro conditions (spheroid culture versus adherent culture) in relation to DC immunotherapy: (1) We studied the specific spheroid-culture proteome and assessed the clinical importance of spheroid proteins. (2) We evaluated the immunogenicity of spheroid lysate – both compared to adherent conditions.
We used seven spheroid culture systems, three of them patient-derived. Stemness-related markers were studied in those three via immunofluorescence. Spheroid-specific protein expression was measured via quantitative proteomics. The Cancer Genome Atlas (TCGA) survival data was used to investigate the clinical impact of spheroid proteins. Immunogenicity of spheroid versus adherent cell lysate was explored in autologous ELISPOT systems (DCs and T cells from the three patients).
(1) The differential proteome of spheroid versus adherent glioblastoma culture conditions could successfully be established. The top 10 identified spheroid-specific proteins were associated with significantly decreased overall survival (TCGA MIT/Harvard cohort; n = 350, P = 0.014). (2) In exploratory experiments, immunogenicity of spheroid lysate vis-á-vis interferon (IFN)γ production was lower than that of adherent cell lysate (IFNγ ELISPOT; P = 0.034).
Spheroid culture proteins seem to represent survival-relevant targets, supporting the use of spheroid culture conditions as an antigen source for DC immunotherapy. However, immunogenicity enhancement should be considered for future research. Transferability of our findings in terms of clinical impact and regarding different spheroid-generation techniques needs further validation.

View all citing articles on Scopus

View full text

Phase I study of tumor Ag-loaded IL-12 secreting semi-mature DC for the treatment of pediatric cancer

Background

Methods

Results

Discussion

Introduction

Section snippets

Patient characteristics

Manufacturing and characterization of the cancer vaccine

Discussion

Acknowledgements

Immunol Today

Cytotherapy

Blood

Blood

Immunobiology of dendritic cells

Annu Rev Immunol

Dendritic cells as adjuvants in antitumor immune therapy

Onkologie

Vaccination of pediatric solid tumor patients with tumor lysate-pulsed dendritic cells can expand specific T cells and mediate tumor regression

Cancer Res

Pilot trial of tumor-specific peptide vaccination and continuous infusion interleukin-2 in patients with recurrent Ewing sarcoma and alveolar rhabdomyosarcoma: an inter-institute NIH study

Med Pediatr Oncol

Generation of tumor immunity by bone marrow-derived dendritic cells correlates with dendritic cell maturation stage

J Immunol

Major histocompatibility class I presentation of soluble antigen facilitated by Mycobacterium tuberculosis infection

Proc Natl Acad Sci USA

MHC class I-restricted cytotoxic lymphocyte responses induced by enterotoxin-based mucosal adjuvants

J Immunol

Immunostimulatory DNA-based vaccines induce cytotoxic lymphocyte activity by a T-helper cell-independent mechanism

Nat Biotechnol

Listeria monocytogenes infection overcomes the requirement for CD40 ligand in exogenous antigen presentation to CD8(+) T cells

J Immunol

Cytotoxic T-cell immunity to virus-infected non-haematopoietic cells requires presentation of exogenous antigen

Nature

Rapid cytotoxic T lymphocyte activation occurs in the draining lymph nodes after cutaneous herpes simplex virus infection as a result of early antigen presentation and not the presence of virus

J Exp Med

Cross-priming of CD8+ T cells stimulated by virus-induced type I interferon

Nat Immunol

A conditioned dendritic cell can be a temporal bridge between a CD4+ T-helper and a T-killer cell

Nature

Help for cytotoxic-T-cell responses is mediated by CD40 signalling

Nature

T-cell help for cytotoxic T lymphocytes is mediated by CD40–CD40L interactions

Nature

Soluble antigen and CD40 triggering are sufficient to induce primary and memory cytotoxic T cells

J Immunol

Dendritic cell lineage, plasticity and cross-regulation

Nat Immunol

Interleukin-12 and the regulation of innate resistance and adaptive immunity

Nat Rev Immunol

TH1 and TH2 cells: different patterns of lymphokine secretion lead to different functional properties

Annu Rev Immunol

Semi-mature IL-12 secreting dendritic cells present exogenous antigen to trigger cytolytic immune responses

Cancer Immunol Immunother

Generation of potent anti-tumor immunity in mice by interleukin-12-secreting dendritic cells

Cancer Immunol Immunother

Cross-priming of CD8⁺ T cells stimulated by virus-induced type I interferon

A conditioned dendritic cell can be a temporal bridge between a CD4⁺ T-helper and a T-killer cell