Exposure to the antimicrobial peptide LL-37 produces dendritic cells optimized for immunotherapy

ABSTRACT Immunization of patients with autologous, ex vivo matured dendritic cell (DC) preparations, in order to prime antitumor T-cell responses, is the focus of intense research. Despite progress and approval of clinical approaches, significant enhancement of these personalized immunotherapies is urgently needed to improve efficacy. We show that immunotherapeutic murine and human DC, generated in the presence of the antimicrobial host defense peptide LL-37, have dramatically enhanced expansion and differentiation of cells with key features of the critical CD103+/CD141+ DC subsets, including enhanced cross-presentation and co-stimulatory capacity, and upregulation of CCR7 with improved migratory capacity. These LL-37-DC enhanced proliferation, activation and cytokine production by CD8+ (but not CD4+) T cells in vitro and in vivo. Critically, tumor antigen-presenting LL-37-DC increased migration of primed, activated CD8+ T cells into established squamous cell carcinomas in mice, and resulted in tumor regression. This advance therefore has the potential to dramatically enhance DC immunotherapy protocols.

[1]  A. Villani,et al.  Large-Scale Human Dendritic Cell Differentiation Revealing Notch-Dependent Lineage Bifurcation and Heterogeneity , 2018, Cell reports.

[2]  Nina Bhardwaj,et al.  Towards superior dendritic-cell vaccines for cancer therapy , 2018, Nature Biomedical Engineering.

[3]  Nina Bhardwaj,et al.  Dendritic cell-based immunotherapy , 2016, Cell Research.

[4]  T. Kaisho,et al.  Critical Role for CD103(+)/CD141(+) Dendritic Cells Bearing CCR7 for Tumor Antigen Trafficking and Priming of T Cell Immunity in Melanoma. , 2016, Cancer cell.

[5]  F. Ginhoux,et al.  Expansion and Activation of CD103(+) Dendritic Cell Progenitors at the Tumor Site Enhances Tumor Responses to Therapeutic PD-L1 and BRAF Inhibition. , 2016, Immunity.

[6]  B. Dréno,et al.  PD-1 expression conditions T cell avidity within an antigen-specific repertoire , 2015, Oncoimmunology.

[7]  M. Dalod,et al.  In Vitro Generation of Human XCR1(+) Dendritic Cells from CD34(+) Hematopoietic Progenitors. , 2016, Methods in molecular biology.

[8]  C. Figdor,et al.  Effective Clinical Responses in Metastatic Melanoma Patients after Vaccination with Primary Myeloid Dendritic Cells , 2015, Clinical Cancer Research.

[9]  Adam Byron,et al.  Nuclear FAK Controls Chemokine Transcription, Tregs, and Evasion of Anti-tumor Immunity , 2015, Cell.

[10]  P. Chakravarty,et al.  GM-CSF Mouse Bone Marrow Cultures Comprise a Heterogeneous Population of CD11c(+)MHCII(+) Macrophages and Dendritic Cells. , 2015, Immunity.

[11]  P. Hart,et al.  Cross-presentation of cutaneous melanoma antigen by migratory XCR1+CD103− and XCR1+CD103+ dendritic cells , 2015, Oncoimmunology.

[12]  N. Bhardwaj,et al.  Cancer immunotherapy: Dendritic-cell vaccines on the move , 2015, Nature.

[13]  M. Merad,et al.  Selective and efficient generation of functional Batf3-dependent CD103+ dendritic cells from mouse bone marrow. , 2014, Blood.

[14]  Sebastian Amigorena,et al.  Dissecting the tumor myeloid compartment reveals rare activating antigen-presenting cells critical for T cell immunity. , 2014, Cancer cell.

[15]  Vivien W. Chan,et al.  Macrophage IL-10 blocks CD8+ T cell-dependent responses to chemotherapy by suppressing IL-12 expression in intratumoral dendritic cells. , 2014, Cancer cell.

[16]  D. Douek,et al.  PD-1 identifies the patient-specific CD8⁺ tumor-reactive repertoire infiltrating human tumors. , 2014, The Journal of clinical investigation.

[17]  H. Mages,et al.  Ontogenic, Phenotypic, and Functional Characterization of XCR1+ Dendritic Cells Leads to a Consistent Classification of Intestinal Dendritic Cells Based on the Expression of XCR1 and SIRPα , 2014, bioRxiv.

[18]  G. Linette,et al.  IL-12p70-producing patient DC vaccine elicits Tc1-polarized immunity. , 2013, The Journal of clinical investigation.

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

[20]  Patrice Ravel,et al.  PD-1-expressing tumor-infiltrating T cells are a favorable prognostic biomarker in HPV-associated head and neck cancer. , 2013, Cancer research.

[21]  Michael Poidinger,et al.  Human Tissues Contain CD141hi Cross-Presenting Dendritic Cells with Functional Homology to Mouse CD103+ Nonlymphoid Dendritic Cells , 2012, Immunity.

[22]  H. Mages,et al.  Expression of XCR1 Characterizes the Batf3-Dependent Lineage of Dendritic Cells Capable of Antigen Cross-Presentation , 2012, Front. Immun..

[23]  V. Brunton,et al.  The role of focal adhesion kinase catalytic activity on the proliferation and migration of squamous cell carcinoma cells , 2012, International journal of cancer.

[24]  K. Murphy,et al.  DNGR-1 is a specific and universal marker of mouse and human Batf3-dependent dendritic cells in lymphoid and nonlymphoid tissues. , 2012, Blood.

[25]  F. Nestle,et al.  Resident CD141 (BDCA3)+ dendritic cells in human skin produce IL-10 and induce regulatory T cells that suppress skin inflammation , 2012, The Journal of experimental medicine.

[26]  C. Sousa.  Faculty Opinions recommendation of Host type I IFN signals are required for antitumor CD8+ T cell responses through CD8{alpha}+ dendritic cells. , 2011 .

[27]  K. Murphy,et al.  Host type I IFN signals are required for antitumor CD8+ T cell responses through CD8α+ dendritic cells , 2011, The Journal of experimental medicine.

[28]  Li Wu,et al.  The Acquisition of Antigen Cross-Presentation Function by Newly Formed Dendritic Cells , 2011, The Journal of Immunology.

[29]  J. Engh,et al.  Induction of CD8+ T-cell responses against novel glioma-associated antigen peptides and clinical activity by vaccinations with {alpha}-type 1 polarized dendritic cells and polyinosinic-polycytidylic acid stabilized by lysine and carboxymethylcellulose in patients with recurrent malignant glioma. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[30]  P. Kantoff,et al.  Sipuleucel-T immunotherapy for castration-resistant prostate cancer. , 2010, The New England journal of medicine.

[31]  P. Rod Dunbar,et al.  Human CD141+ (BDCA-3)+ dendritic cells (DCs) represent a unique myeloid DC subset that cross-presents necrotic cell antigens , 2010, The Journal of experimental medicine.

[32]  J. Villadangos,et al.  Found in translation: the human equivalent of mouse CD8+ dendritic cells , 2010, The Journal of experimental medicine.

[33]  K. Murphy,et al.  Peripheral CD103+ dendritic cells form a unified subset developmentally related to CD8α+ conventional dendritic cells , 2010, The Journal of experimental medicine.

[34]  B. Malissen,et al.  Comparative genomics as a tool to reveal functional equivalences between human and mouse dendritic cell subsets , 2010, Immunological reviews.

[35]  J. Rodriguez-Barbosa,et al.  Development and functional specialization of CD103+ dendritic cells , 2010, Immunological reviews.

[36]  F. Ginhoux,et al.  The origin and development of nonlymphoid tissue CD103+ DCs , 2009, The Journal of experimental medicine.

[37]  Andreas Hutloff,et al.  Selective expression of the chemokine receptor XCR1 on cross-presenting dendritic cells determines cooperation with CD8+ T cells. , 2009, Immunity.

[38]  E. Unanue,et al.  Batf3 Deficiency Reveals a Critical Role for CD8α+ Dendritic Cells in Cytotoxic T Cell Immunity , 2008, Science.

[39]  Eric Vivier,et al.  Novel insights into the relationships between dendritic cell subsets in human and mouse revealed by genome-wide expression profiling , 2008, Genome Biology.

[40]  T. S. Wilkinson,et al.  The human cationic host defense peptide LL‐37 mediates contrasting effects on apoptotic pathways in different primary cells of the innate immune system , 2006, Journal of leukocyte biology.

[41]  Li Wu,et al.  Cutting Edge: Generation of Splenic CD8+ and CD8− Dendritic Cell Equivalents in Fms-Like Tyrosine Kinase 3 Ligand Bone Marrow Cultures1 , 2005, The Journal of Immunology.

[42]  C. Figdor,et al.  Migration of dendritic cell based cancer vaccines: in vivo veritas? , 2005, Current opinion in immunology.

[43]  J. Kirkwood,et al.  alpha-type-1 polarized dendritic cells: a novel immunization tool with optimized CTL-inducing activity. , 2004, Cancer research.

[44]  R. Hancock,et al.  The Human Cationic Peptide LL-37 Induces Activation of the Extracellular Signal-Regulated Kinase and p38 Kinase Pathways in Primary Human Monocytes1 , 2004, The Journal of Immunology.

[45]  Robert E. W. Hancock,et al.  The Cationic Antimicrobial Peptide LL-37 Modulates Dendritic Cell Differentiation and Dendritic Cell-Induced T Cell Polarization1 , 2004, The Journal of Immunology.

[46]  W. Oyen,et al.  Effective migration of antigen-pulsed dendritic cells to lymph nodes in melanoma patients is determined by their maturation state. , 2003, Cancer research.

[47]  N. Bhardwaj,et al.  A clinical grade cocktail of cytokines and PGE2 results in uniform maturation of human monocyte-derived dendritic cells: implications for immunotherapy. , 2002, Vaccine.

[48]  M. Peshwa,et al.  Immunotherapy of hormone-refractory prostate cancer with antigen-loaded dendritic cells. , 2000, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[49]  S. Miltenyi,et al.  BDCA-2, BDCA-3, and BDCA-4: Three Markers for Distinct Subsets of Dendritic Cells in Human Peripheral Blood , 2000, The Journal of Immunology.

[50]  C. Maliszewski,et al.  Generation of murine dendritic cells from flt3-ligand-supplemented bone marrow cultures. , 2000, Blood.

[51]  T. Raife,et al.  Thrombomodulin expression by human keratinocytes. Induction of cofactor activity during epidermal differentiation. , 1994, The Journal of clinical investigation.