Intranodal vaccination with mRNA-optimized dendritic cells in metastatic melanoma patients

Autologous dendritic cell (DC) therapy is an experimental cellular immunotherapy that is safe and immunogenic in patients with advanced melanoma. In an attempt to further improve the therapeutic responses, we treated 15 patients with melanoma, with autologous monocyte-derived immature DC electroporated with mRNA encoding CD40 ligand (CD40L), CD70 and a constitutively active TLR4 (caTLR4) together with mRNA encoding a tumor-associated antigen (TAA; respectively gp100 or tyrosinase). In addition, DC were pulsed with keyhole limpet hemocyanin (KLH) that served as a control antigen. Production of this DC vaccine with high cellular viability, high expression of co-stimulatory molecules and MHC class I and II and production of IL-12p70, was feasible in all patients. A vaccination cycle consisting of three vaccinations with up to 15×106 DC per vaccination at a biweekly interval, was repeated after 6 and 12 months in the absence of disease progression. mRNA-optimized DC were injected intranodally, because of low CCR7 expression on the DC, and induced de novo immune responses against control antigen. T cell responses against tyrosinase were detected in the skin-test infiltrating lymphocytes (SKIL) of two patients. One mixed tumor response and two durable tumor stabilizations were observed among 8 patients with evaluable disease at baseline. In conclusion, autologous mRNA-optimized DC can be safely administered intranodally to patients with metastatic melanoma but showed limited immunological responses against tyrosinase and gp100.

[1]  C. Figdor,et al.  Favorable overall survival in stage III melanoma patients after adjuvant dendritic cell vaccination , 2016, Oncoimmunology.

[2]  Michael R Stratton,et al.  High-throughput epitope discovery reveals frequent recognition of neo-antigens by CD4+ T cells in human melanoma , 2014, Nature Medicine.

[3]  B. Neyns,et al.  Phase II study of autologous mRNA electroporated dendritic cells (TriMixDC-MEL) in combination with Ipilimumab in patients with pretreated advanced melanoma , 2014, Journal of Immunotherapy for Cancer.

[4]  B. Neyns,et al.  Phase II study of autologous mRNA electroporated dendritic cells (TriMixDC-MEL) in combination with ipilimumab in patients with pretreated advanced melanoma. , 2014 .

[5]  B. Neyns,et al.  A phase IB study on intravenous synthetic mRNA electroporated dendritic cell immunotherapy in pretreated advanced melanoma patients. , 2013, Annals of oncology : official journal of the European Society for Medical Oncology.

[6]  C. Figdor,et al.  Naturally circulating dendritic cells to vaccinate cancer patients , 2013, Oncoimmunology.

[7]  W. Oyen,et al.  Natural human plasmacytoid dendritic cells induce antigen-specific T-cell responses in melanoma patients. , 2013, Cancer research.

[8]  A. Croockewit,et al.  Targeting CD4(+) T-helper cells improves the induction of antitumor responses in dendritic cell-based vaccination. , 2013, Cancer research.

[9]  K. Thielemans,et al.  Overcoming HLA restriction in clinical trials , 2012, Oncoimmunology.

[10]  C. Figdor,et al.  Skin-test infiltrating lymphocytes early predict clinical outcome of dendritic cell-based vaccination in metastatic melanoma. , 2012, Cancer research.

[11]  C. Figdor,et al.  Vaccination with mRNA-Electroporated Dendritic Cells Induces Robust Tumor Antigen-Specific CD4+ and CD8+ T Cells Responses in Stage III and IV Melanoma Patients , 2012, Clinical Cancer Research.

[12]  B. Neyns,et al.  Dendritic cells loaded with mRNA encoding full-length tumor antigens prime CD4+ and CD8+ T cells in melanoma patients. , 2012, Molecular therapy : the journal of the American Society of Gene Therapy.

[13]  C. Punt,et al.  Humoral anti-KLH responses in cancer patients treated with dendritic cell-based immunotherapy are dictated by different vaccination parameters , 2012, Cancer Immunology, Immunotherapy.

[14]  B. Neyns,et al.  Epitope and HLA-type independent monitoring of antigen-specific T-cells after treatment with dendritic cells presenting full-length tumor antigens. , 2012, Journal of immunological methods.

[15]  W. Oyen,et al.  Route of Administration Modulates the Induction of Dendritic Cell Vaccine–Induced Antigen-Specific T Cells in Advanced Melanoma Patients , 2011, Clinical Cancer Research.

[16]  B. Neyns,et al.  Therapeutic Vaccination With an Autologous mRNA Electroporated Dendritic Cell Vaccine in Patients With Advanced Melanoma , 2011, Journal of immunotherapy.

[17]  C. Figdor,et al.  Wild-type and modified gp100 peptide-pulsed dendritic cell vaccination of advanced melanoma patients can lead to long-term clinical responses independent of the peptide used , 2010, Cancer Immunology, Immunotherapy.

[18]  B. Neyns,et al.  Single-Step Antigen Loading and Activation of Dendritic Cells by mRNA Electroporation for the Purpose of Therapeutic Vaccination in Melanoma Patients , 2009, Clinical Cancer Research.

[19]  W. Oyen,et al.  In situ expression of tumor antigens by messenger RNA-electroporated dendritic cells in lymph nodes of melanoma patients. , 2009, Cancer research.

[20]  C. Figdor,et al.  Polyinosinic polycytidylic acid prevents efficient antigen expression after mRNA electroporation of clinical grade dendritic cells , 2009, Cancer Immunology, Immunotherapy.

[21]  B. Neyns,et al.  Enhancing the T-cell stimulatory capacity of human dendritic cells by co-electroporation with CD40L, CD70 and constitutively active TLR4 encoding mRNA. , 2008, Molecular therapy : the journal of the American Society of Gene Therapy.

[22]  C. Figdor,et al.  Dendritic cell vaccines in melanoma: from promise to proof? , 2008, Critical reviews in oncology/hematology.

[23]  R. Steinman,et al.  Taking dendritic cells into medicine , 2007, Nature.

[24]  W. Oyen,et al.  Development of 111In-labeled tumor-associated antigen peptides for monitoring dendritic-cell-based vaccination. , 2006, Nuclear medicine and biology.

[25]  J. Rodríguez-Fernández,et al.  The Multiple Personalities of the Chemokine Receptor CCR7 in Dendritic Cells1 , 2006, The Journal of Immunology.

[26]  C. Figdor,et al.  Immunomonitoring tumor-specific T cells in delayed-type hypersensitivity skin biopsies after dendritic cell vaccination correlates with clinical outcome. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[27]  J. Borst,et al.  CD27 and CD70 in T cell and B cell activation. , 2005, Current opinion in immunology.

[28]  E. Gilboa,et al.  Induction of Human Dendritic Cell Maturation Using Transfection with RNA Encoding a Dominant Positive Toll-Like Receptor 41 , 2004, The Journal of Immunology.

[29]  C. Figdor,et al.  Dendritic cell immunotherapy: mapping the way , 2004, Nature Medicine.

[30]  G. Schuler,et al.  Functional Analysis of Tumor-Specific Th Cell Responses Detected in Melanoma Patients after Dendritic Cell-Based Immunotherapy1 , 2004, The Journal of Immunology.

[31]  E. Gilboa,et al.  Injection of Immature Dendritic Cells into Adjuvant-Treated Skin Obviates the Need for Ex Vivo Maturation 1 , 2003, The Journal of Immunology.

[32]  C. Figdor,et al.  Maturation of dendritic cells is a prerequisite for inducing immune responses in advanced melanoma patients. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[33]  M. Grégoire,et al.  Optimizing dendritic cell–based anticancer immunotherapy: maturation state does have clinical impact , 2003, Cancer Immunology, Immunotherapy.

[34]  G. Trinchieri,et al.  Interleukin-12 and the regulation of innate resistance and adaptive immunity , 2003, Nature Reviews Immunology.

[35]  A. Enk,et al.  A comparison of two types of dendritic cell as adjuvants for the induction of melanoma‐specific T‐cell responses in humans following intranodal injection , 2001, International journal of cancer.

[36]  C. van Broeckhoven,et al.  Highly efficient gene delivery by mRNA electroporation in human hematopoietic cells: superiority to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for tumor antigen loading of dendritic cells. , 2001, Blood.

[37]  Forest M. White,et al.  Phosphorylated Peptides Are Naturally Processed and Presented by Major Histocompatibility Complex Class I Molecules in Vivo , 2000, The Journal of experimental medicine.

[38]  D. Mitchell,et al.  RNA-transfected dendritic cells in cancer immunotherapy. , 2000, The Journal of clinical investigation.

[39]  R. Crystal,et al.  Dendritic cells modified to express CD40 ligand elicit therapeutic immunity against preexisting murine tumors. , 2000, Blood.

[40]  P. Ricciardi-Castagnoli,et al.  Dendritic cells directly trigger NK cell functions: Cross-talk relevant in innate anti-tumor immune responses in vivo , 1999, Nature Medicine.

[41]  R. Steinman,et al.  Dendritic cells and the control of immunity , 1998, Nature.

[42]  T. Mcclanahan,et al.  A dendritic-cell-derived C–C chemokine that preferentially attracts naive T cells , 1997, Nature.

[43]  J. Shabanowitz,et al.  An HLA-A2-restricted tyrosinase antigen on melanoma cells results from posttranslational modification and suggests a novel pathway for processing of membrane proteins , 1996, The Journal of experimental medicine.