CD40-targeted adenoviral gene transfer to dendritic cells through the use of a novel bispecific single-chain Fv antibody enhances cytotoxic T cell activation.
Adenoviral (Ad) transduction of dendritic cells (DC) is a promising vaccination strategy. However, clinical applicability of Ad vectors is hampered by the necessity to use high titers of infectious Ad particles for efficient DC transduction. Here, we report on the production of a bacterially expressed bispecific conjugate, consisting of a fusion of recombinant single-chain (sc) mAb Fv fragments, which bind and neutralize the Ad fiber knob (through the S11 mAb scFv) and retarget Ad to CD40 on the DC surface (through the G28-5 mAb scFv). We show that this bispecific scFv fusion protein significantly enhances transduction efficiency of monocyte-derived DC (MoDC), reduces the amount of virus needed for a given level of transduction, and increases the ability of MoDC to activate CTL in an antigen specific manner. This single-component conjugate may prove to be a valuable immunotherapeutic tool for the targeting of Ad to DC in vivo.
Brandão
Joana G
JG
Department of Pathology, VU University Medical Center, PO Box 7057, 1007 Amsterdam MB, The Netherlands.
Scheper
Rik J
RJ
Lougheed
Sinéad M
SM
Curiel
David T
DT
Tillman
Bryan W
BW
Gerritsen
Winald R
WR
van den Eertwegh
Alfons J M
AJ
Pinedo
Herbert M
HM
Haisma
Hidde J
HJ
de Gruijl
Tanja D
TD
D000256:Adenoviridae / Q000235:genetics* / Q000276:immunology*; D000818:Animals; D018033:Antibodies, Bispecific / Q000276:immunology*; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D019013:CD40 Antigens / Q000235:genetics / Q000276:immunology*; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D017931:DNA Primers; D003713:Dendritic Cells / Q000276:immunology* / Q000821:virology; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D008164:Luminescent Proteins / Q000235:genetics / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D018033:Antibodies, Bispecific; D000911:Antibodies, Monoclonal; D019013:CD40 Antigens; D017931:DNA Primers; D007074:Immunoglobulin G; D007135:Immunoglobulin Variable Region; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins