Blood dendritic cells generated with Flt3 ligand and CD40 ligand prime CD8+ T cells efficiently in cancer patients.
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.
Davis
Ian D
ID
Ludwig Institute for Cancer Research, Austin Health, Studley Road, Heidelberg, Victoria 3084, Australia. Ian.Davis@ludwig.edu.au
Chen
Qiyuan
Q
Morris
Leone
L
Quirk
Juliet
J
Stanley
Maureen
M
Tavarnesi
Maria L
ML
Parente
Phillip
P
Cavicchiolo
Tina
T
Hopkins
Wendie
W
Jackson
Heather
H
Dimopoulos
Nektaria
N
Tai
Tsin Yee
TY
MacGregor
Duncan
D
Browning
Judy
J
Svobodova
Suzanne
S
Caron
Dania
D
Maraskovsky
Eugene
E
Old
Lloyd J
LJ
Chen
Weisan
W
Cebon
Jonathan
J
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*
D016827:CD8 Antigens; D019496:Cancer Vaccines; D008565:Membrane Proteins; D010455:Peptides; D011948:Receptors, Antigen, T-Cell; C084881:flt3 ligand protein; D023201:CD40 Ligand