title: A versatile bifunctional dendritic cell targeting vaccine vector.
abstract: We have developed an efficient versatile in vivo dendritic cell (DC) targeting vector for delivering different classes of antigens such as proteins, peptide, glycolipids and naked DNA for vaccine applications. A single chain antibody (scFv) that recognizes DEC-205 receptor of DC was fused with a core-streptavidin domain and expressed in Escherichia coli using the T7 expression system. The bifunctional fusion protein (bfFp) was expressed as a periplasmic soluble protein and affinity-purified in its monomeric form. The bifunctional activity against DEC-205 and biotin was characterized by ELISA and Western blot. In vivo DC targeting of a diverse group of biotinylated antigens such as viral and bacterial proteins, a cancer peptide, gangliosides and DNA of certain infectious diseases was conducted in mice. Results show that in the presence of bfFp and costimulatory anti-CD40 mAb, both humoral and cell-mediated responses were augmented in either the single antigen or multiple antigen targeting strategy. Lastly, bfFp based DC targeting of antigens in low doses may be a useful strategy for the design of monovalent or polyvalent vaccines for the masses.
authors.affiliation: University of Alberta, Edmonton, Alberta, Canada T6G 2N8.
authors.forename: Welson W
authors.initials: WW
authors.lastname: Wang
authors.forename: Dipankar
authors.initials: D
authors.lastname: Das
authors.forename: Mavanur R
authors.initials: MR
authors.lastname: Suresh
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D001428:Bacterial Vaccines / Q000235:genetics / Q000378:metabolism*; D001710:Biotin; D002460:Cell Line; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D006241:Haptens / Q000276:immunology; D007371:Interferon-gamma / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014765:Viral Vaccines / Q000235:genetics / Q000378:metabolism*
chemical_list: D000906:Antibodies; D001428:Bacterial Vaccines; D006241:Haptens; D011994:Recombinant Proteins; D014765:Viral Vaccines; D001710:Biotin; D007371:Interferon-gamma