title: CMRF-56(+) blood dendritic cells loaded with mRNA induce effective antigen-specific cytotoxic T-lymphocyte responses.
abstract: There are numerous transcriptional, proteomic and functional differences between monocyte-derived dendritic cells (Mo-DC) and primary blood dendritic cells (BDC). The CMRF-56 monoclonal antibody (mAb) recognizes a cell surface marker, which is upregulated on BDC following overnight culture. Given its unique ability to select a heterogeneous population of BDC, we engineered a human chimeric (h)CMRF-56 IgG4 mAb to isolate primary BDC for potential therapeutic vaccination. The ability to select multiple primary BDC subsets from patients and load them with in vitro transcribed (IVT) mRNA encoding tumor antigen might circumvent the issues limiting the efficacy of Mo-DC. After optimizing and validating the purification of hCMRF-56(+) BDC, we showed that transfection of hCMRF-56(+) BDC with mRNA resulted in efficient mRNA translation and antigen presentation by myeloid BDC subsets, while preserving superior DC functions compared to Mo-DC. Immune selected and transfected hCMRF-56(+) BDC migrated very efficiently in vitro and as effectively as cytokine matured Mo-DC in vivo. Compared to Mo-DC, hCMRF-56(+) BDC transfected with influenza matrix protein M1 displayed superior MHC peptide presentation and generated potent antigen specific CD8(+) T-cell recall responses, while Wilms tumor 1 (WT1) transfected CMRF-56(+) BDC generated effective primary autologous cytotoxic T-cell responses. The ability of the combined DC subsets within hCMRF-56(+) BDC to present mRNA delivered tumor antigens merits phase I evaluation as a reproducible generic platform for the next generation of active DC immune therapies.
authors.affiliation: ANZAC Research Institute, Concord, NSW, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW, Australia.
authors.forename: Phillip D
authors.initials: PD
authors.lastname: Fromm
authors.affiliation: ANZAC Research Institute, Concord, NSW, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW, Australia.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Papadimitrious
authors.affiliation: ANZAC Research Institute , Concord, NSW, Australia.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Hsu
authors.affiliation: ANZAC Research Institute, Concord, NSW, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW, Australia.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Van Kooten Losio
authors.affiliation: ANZAC Research Institute, Concord, NSW, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW, Australia.
authors.forename: Nirupama D
authors.initials: ND
authors.lastname: Verma
authors.affiliation: ANZAC Research Institute, Concord, NSW, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW, Australia.
authors.forename: Tsun Ho
authors.initials: TH
authors.lastname: Lo
authors.affiliation: ANZAC Research Institute, Concord, NSW, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW, Australia.
authors.forename: Pablo A
authors.initials: PA
authors.lastname: Silveira
authors.affiliation: ANZAC Research Institute, Concord, NSW, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW, Australia.
authors.forename: Christian E
authors.initials: CE
authors.lastname: Bryant
authors.affiliation: Program in Immunology, Fred Hutchinson Cancer Research Center , Seattle, WA, USA.
authors.forename: Cameron J
authors.initials: CJ
authors.lastname: Turtle
authors.affiliation: Mater Medical Research Institute , Raymond Terrace, QLD, Australia.
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Prue
authors.affiliation: Mater Medical Research Institute , Raymond Terrace, QLD, Australia.
authors.forename: Peter
authors.initials: P
authors.lastname: Vukovic
authors.affiliation: Mater Medical Research Institute , Raymond Terrace, QLD, Australia.
authors.forename: David J
authors.initials: DJ
authors.lastname: Munster
authors.affiliation: Mater Medical Research Institute , Raymond Terrace, QLD, Australia.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Nagasaki
authors.affiliation: School of Chemistry and Molecular Biosciences, University of Queensland , St Lucia, QLD, Australia.
authors.forename: Ross T
authors.initials: RT
authors.lastname: Barnard
authors.affiliation: ANZAC Research Institute , Concord, NSW, Australia.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Mahler
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, University of Queensland , St Lucia, QLD, Australia.
authors.forename: Sébastien A
authors.initials: SA
authors.lastname: Anguille
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, University of Queensland , St Lucia, QLD, Australia.
authors.forename: Zwi
authors.initials: Z
authors.lastname: Berneman
authors.affiliation: Antwerp University Hospital, Center for Cell Therapy and Regenerative Medicine, Antwerp, Belgium; Department of Tissue Pathology and Diagnostic Oncology, Royal Prince Alfred Hospital, Camperdown, NSW, Australia; The Kinghorn Cancer Center/Garvan Institute of Medical Research, Darlinghurst, NSW, Australia.
authors.forename: Lisa G
authors.initials: LG
authors.lastname: Horvath
authors.affiliation: ANZAC Research Institute, Concord, NSW, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW, Australia; Chris O'Brien Lifehouse, Department of Medical Oncology, Royal Prince Alfred Hospital, Camperdown, NSW, Australia.
authors.forename: Kenneth F
authors.initials: KF
authors.lastname: Bradstock
authors.affiliation: Sydney Medical School, University of Sydney, Camperdown, NSW, Australia; Haematology Department, Westmead Hospital, Westmead, NSW, Australia.
authors.forename: Douglas E
authors.initials: DE
authors.lastname: Joshua
authors.affiliation: ANZAC Research Institute, Concord, NSW, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW, Australia.
authors.forename: Georgina J
authors.initials: GJ
authors.lastname: Clark
authors.affiliation: ANZAC Research Institute, Concord, NSW, Australia; Sydney Medical School, University of Sydney, Camperdown, NSW, Australia; Department of Haematology, Royal Prince Alfred Hospital, Camperdown, NSW, Australia; Department of Haematology, Concord Repatriation General Hospital, Concord, NSW, Australia.
authors.forename: Derek N J
authors.initials: DN
authors.lastname: Hart
keywords: Antigen presentation; CTL; immunoselection; mRNA transfection; therapeutic DC vaccination