title: Potent maturation of monocyte-derived dendritic cells after CD40L lentiviral gene delivery.
abstract: Dendritic cells (DCs) are being evaluated in immunization protocols to enhance immunity against infectious diseases and cancer. Interaction of T-helper cells expressing CD40 ligand (CD40L) with its cognate CD40 receptor on DCs leads to a mature DC phenotype, characterized by increased capacity of antigen presentation to cytotoxic T cells. The authors examined the ability of third-generation self-inactivating lentiviral vectors expressing CD40L to induce autonomous maturation of ex vivo expanded human monocyte-derived dendritic cells. Transduction with lentiviral vectors achieved a highly efficient gene transfer of CD40L to DCs, which correlated with phenotypic maturation as shown by the expression of immunologic relevant markers (CD83, CD80, MHCI) and secretion of IL-12, whereas DC phenotype was not affected by a control vector expressing only the green fluorescent protein marker. Addition of recombinant IFN-gamma to DCs at the time of CD40L transduction further enhanced IL-12 production, and when co-cultured with allogeneic and autologous CD8+ and CD4+ T cells, a potent activation was observed. Autologous responses against an HLA-A2-restricted influenza peptide (Flu-M1) and a tumor-associated antigenic peptide (gp100 210M) were significantly enhanced when CD40L transduced DCs were used as antigen-presenting cells for in vitro stimulation of CD8+ cytotoxic T lymphocytes. These results demonstrate that endogenous expression of CD40L by lentivirally transduced DCs induced their autonomous maturation to a phenotype comparable to that induced by optimal concentrations of soluble CD40L, providing a novel tool for genetic manipulation of DCs.
authors.affiliation: Institute for Genetic Medicine, Keck School of Medicine, University of Southern California, Los Angeles, California, USA.
authors.forename: Richard C
authors.initials: RC
authors.lastname: Koya
authors.forename: Nori
authors.initials: N
authors.lastname: Kasahara
authors.forename: Patricia M B
authors.initials: PM
authors.lastname: Favaro
authors.forename: Roy
authors.initials: R
authors.lastname: Lau
authors.forename: Huy Q
authors.initials: HQ
authors.lastname: Ta
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Weber
authors.forename: Renata
authors.initials: R
authors.lastname: Stripecke
mesh_terms: D000911:Antibodies, Monoclonal; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D023201:CD40 Ligand / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D002460:Cell Line; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D016130:Immunophenotyping; D018664:Interleukin-12 / Q000096:biosynthesis; D016086:Lentivirus / Q000235:genetics*; D009000:Monocytes; D011994:Recombinant Proteins; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D023201:CD40 Ligand; D018664:Interleukin-12; D016178:Granulocyte-Macrophage Colony-Stimulating Factor