title: Blood dendritic cells generated with Flt3 ligand and CD40 ligand prime CD8+ T cells efficiently in cancer patients.
abstract: Flt3 ligand mobilizes dendritic cells (DCs) into blood, allowing generation in vivo of large numbers of DCs for immunotherapy. These immature DCs can be rapidly activated by soluble CD40 ligand (CD40L). We developed a novel overnight method using these cytokines to produce DCs for cancer immunotherapy. Flt3 ligand-mobilized DCs (FLDCs) were isolated, activated with CD40L, loaded with antigenic peptides from influenza matrix protein, hepatitis B core antigen, NY-ESO-1, MAGE-A4, and MAGE-A10, and injected into patients with resected melanoma. Three injections were given at 4-week intervals. Study end points included antigen-specific immune responses (skin reactions to peptides alone or peptide-pulsed FLDCs; circulating T-cell responses), safety, and toxicity. No patient had a measurable tumor. Six patients were entered. FLDCs were obtained, enriched, and cultured under Good Manufacturing Practice grade conditions. Overnight culture with soluble CD40L caused marked up-regulation of activation markers (CD83 and HLA-DR). These FLDCs were functional and able to stimulate antigen-specific T cells in vitro. No significant adverse events were attributable to FLDCs. Peptide-pulsed FLDCs caused strong local skin reactions up to 60 mm diameter with intense perivascular infiltration of T cells, exceeding those seen in our previous peptide-based protocols. Antigen-specific blood T-cell responses were induced, including responses to an antigen for which the patients were naive (hepatitis B core antigen) and MAGE-A10. MAGE-A10-specific T cells with a skewed T-cell receptor repertoire were detected in 1 patient in blood ex vivo and from tumor biopsies. Vaccination with FLDCs pulsed with peptides is safe and primes immune responses to cancer antigens.
authors.affiliation: Ludwig Institute for Cancer Research, Austin Health, Studley Road, Heidelberg, Victoria 3084, Australia. Ian.Davis@ludwig.edu.au
authors.forename: Ian D
authors.initials: ID
authors.lastname: Davis
authors.forename: Qiyuan
authors.initials: Q
authors.lastname: Chen
authors.forename: Leone
authors.initials: L
authors.lastname: Morris
authors.forename: Juliet
authors.initials: J
authors.lastname: Quirk
authors.forename: Maureen
authors.initials: M
authors.lastname: Stanley
authors.forename: Maria L
authors.initials: ML
authors.lastname: Tavarnesi
authors.forename: Phillip
authors.initials: P
authors.lastname: Parente
authors.forename: Tina
authors.initials: T
authors.lastname: Cavicchiolo
authors.forename: Wendie
authors.initials: W
authors.lastname: Hopkins
authors.forename: Heather
authors.initials: H
authors.lastname: Jackson
authors.forename: Nektaria
authors.initials: N
authors.lastname: Dimopoulos
authors.forename: Tsin Yee
authors.initials: TY
authors.lastname: Tai
authors.forename: Duncan
authors.initials: D
authors.lastname: MacGregor
authors.forename: Judy
authors.initials: J
authors.lastname: Browning
authors.forename: Suzanne
authors.initials: S
authors.lastname: Svobodova
authors.forename: Dania
authors.initials: D
authors.lastname: Caron
authors.forename: Eugene
authors.initials: E
authors.lastname: Maraskovsky
authors.forename: Lloyd J
authors.initials: LJ
authors.lastname: Old
authors.forename: Weisan
authors.initials: W
authors.lastname: Chen
authors.forename: Jonathan
authors.initials: J
authors.lastname: Cebon
mesh_terms: D000328:Adult; D017951:Antigen Presentation; D023201:CD40 Ligand / Q000276:immunology*; D016827:CD8 Antigens / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D019496:Cancer Vaccines / Q000009:adverse effects / Q000276:immunology / Q000627:therapeutic use*; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D003713:Dendritic Cells / Q000276:immunology*; D005260:Female; D006801:Humans; D016219:Immunotherapy, Adoptive; D007937:Leukapheresis; D008213:Lymphocyte Activation; D008297:Male; D008545:Melanoma / Q000276:immunology / Q000628:therapy*; D008565:Membrane Proteins / Q000276:immunology*; D008875:Middle Aged; D010455:Peptides / Q000276:immunology / Q000627:therapeutic use; D010865:Pilot Projects; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D012878:Skin Neoplasms / Q000276:immunology / Q000628:therapy*
chemical_list: D016827:CD8 Antigens; D019496:Cancer Vaccines; D008565:Membrane Proteins; D010455:Peptides; D011948:Receptors, Antigen, T-Cell; C084881:flt3 ligand protein; D023201:CD40 Ligand