title: Design, expression, and characterization of a novel dendritic cell-targeted proteins.
abstract: In vivo approaches to inducing an effective immune response focus on targeted antigen (Ag) delivery to dendritic cells (DCs). In this study, we developed a new method of targeting plasmid DNA and/or the antigen (Ag)-antibody (Ab) complex to DCs via the DC receptor DEC-205, also known as cluster of differentiation CD205. We cloned and expressed a recombinant protein composed of mouse DEC-205-specific single-chain fragment variable region (mDEC-205-scFv), the streptococcal protein G (SPG) IgG-binding domain and cationic peptide (CP), which named mDEC205-scFv-SPG-CP (msSC). In vitro, the recombinant protein msSC can specifically bind to DCs through the section of mDEC-205-scFv, and bound the Ag-Ab complex via SPG as well as plasmid DNA through electrostatic bonding with CP in vitro. In addition, msSC functioned in a manner similar to anti-DEC-205 monoclonal Ab and bound to mouse bone marrow-derived DCs. It was demonstrated in vivo that msSC can target plasmid DNA to DCs, resulting in efficient uptake and expression. Moreover, msSC can form a complex with pGL3-CMV and transport it to draining lymph nodes when injected in vivo. These results indicate that msSC can be used as a carrier protein for vaccine delivery to DCs via formation of plasmid DNA-Ag-Ab ternary complexes.
authors.affiliation: Beijing Institute of Basic Medical Sciences, 27 Tai Ping Road, Beijing 100850, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Beijing Institute of Basic Medical Sciences, 27 Tai Ping Road, Beijing 100850, China.
authors.forename: Xiao Ming
authors.initials: XM
authors.lastname: Zhu
authors.affiliation: Beijing Institute of Basic Medical Sciences, 27 Tai Ping Road, Beijing 100850, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.affiliation: Beijing Institute of Basic Medical Sciences, 27 Tai Ping Road, Beijing 100850, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Wu
authors.affiliation: Beijing Institute of Basic Medical Sciences, 27 Tai Ping Road, Beijing 100850, China.
authors.forename: Zi Cheng
authors.initials: ZC
authors.lastname: Wang
authors.affiliation: Beijing Institute of Basic Medical Sciences, 27 Tai Ping Road, Beijing 100850, China.
authors.forename: Zhi Yan
authors.initials: ZY
authors.lastname: Du
authors.affiliation: Beijing Institute of Basic Medical Sciences, 27 Tai Ping Road, Beijing 100850, China. Electronic address: yanjinqi@sohu.com.
authors.forename: Jin Qi
authors.initials: JQ
authors.lastname: Yan
authors.affiliation: Beijing Institute of Basic Medical Sciences, 27 Tai Ping Road, Beijing 100850, China. Electronic address: yujyun@126.com.
authors.forename: Ji Yun
authors.initials: JY
authors.lastname: Yu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000276:immunology; D000941:Antigens / Q000276:immunology; D002461:Cell Line, Transformed; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data
chemical_list: D000906:Antibodies; D000936:Antigen-Antibody Complex; D000941:Antigens
keywords: Antigen–antibody complex; DEC-205; Dendritic cell-targeting; Plasmid DNA