title: Targeting of DEC-205 on human dendritic cells results in efficient MHC class II-restricted antigen presentation.
abstract: The use of dendritic cells (DCs) in therapeutic cancer vaccination requires their loading with tumor-specific antigen(s). DEC-205, a phagocytosis receptor mediating antigen uptake, is associated with CD8(+) T-cell responses in mice. Here we fused an anti-DEC-205scFv to an HLA-DP4-restricted epitope from the tumor antigen MAGE-A3, and examined the suitability and efficacy of DEC-205 to deliver a helper epitope to human monocyte-derived DCs (moDCs). The construct specifically bound DEC-205 on human moDCs without negative impact on DC phenotype and function. We measured antigen presentation with specific autologous CD4(+) T cells, generated by TCR-RNA transfection. DEC-205 targeting resulted in significant major histocompatibility complex class II-restricted antigen presentation, and was superior to loading DCs by electroporation of mRNA encoding endosome-targeted MAGE-A3-DCLAMP or by direct peptide pulsing. Anti-DEC-205scFv-MAGE-A3 was presented 100 times more efficiently than the control constructs. DC maturation before or during incubation with anti-DEC-205scFv-MAGE-A3 reduced the interleukin-10/interleukin-2 ratio. Moreover, we successfully applied the DEC-205 targeting strategy to moDCs from malignant melanoma patients. Again, DEC-205-targeted mature DCs (mDCs) presented the antigen more efficiently than peptide-pulsed DCs and maintained their stimulatory capacity after cryoconservation. Thus, DEC-205 targeting represents a feasible and effective method to deliver helper epitopes to DCs in anticancer vaccine strategies, which may also be suitable for DC targeting in vivo.
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Hartmannstrasse 14, Erlangen, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Birkholz
authors.forename: Michael
authors.initials: M
authors.lastname: Schwenkert
authors.forename: Christian
authors.initials: C
authors.lastname: Kellner
authors.forename: Stefanie
authors.initials: S
authors.lastname: Gross
authors.forename: Georg
authors.initials: G
authors.lastname: Fey
authors.forename: Beatrice
authors.initials: B
authors.lastname: Schuler-Thurner
authors.forename: Gerold
authors.initials: G
authors.lastname: Schuler
authors.forename: Niels
authors.initials: N
authors.lastname: Schaft
authors.forename: Jan
authors.initials: J
authors.lastname: Dörrie
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies / Q000008:administration & dosage / Q000235:genetics; D017951:Antigen Presentation / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology* / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D002454:Cell Differentiation / Q000276:immunology; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D000939:Epitopes / Q000008:administration & dosage / Q000235:genetics; D006682:HLA-DP Antigens / Q000378:metabolism; D059868:HLA-DP beta-Chains; D000949:Histocompatibility Antigens Class II / Q000378:metabolism*; D006801:Humans; D016233:Immunotherapy, Active; D066298:In Vitro Techniques; D037181:Lectins, C-Type / Q000276:immunology* / Q000378:metabolism; D008545:Melanoma / Q000276:immunology / Q000628:therapy; D015778:Minor Histocompatibility Antigens; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology; D011956:Receptors, Cell Surface / Q000276:immunology* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D000951:Antigens, Neoplasm; D016207:Cytokines; C093569:DEC-205 receptor; D000939:Epitopes; D006682:HLA-DP Antigens; D059868:HLA-DP beta-Chains; C056284:HLA-DPw4 antigen; D000949:Histocompatibility Antigens Class II; D037181:Lectins, C-Type; C072013:MAGEA3 protein, human; D015778:Minor Histocompatibility Antigens; D009363:Neoplasm Proteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins