Engineered hepatitis B virus surface antigen L protein particles for in vivo active targeting of splenic dendritic cells

Abstract

Hidenori Matsuo,1 Nobuo Yoshimoto,1 Masumi Iijima,1 Tomoaki Niimi,1 Joohee Jung,2,3 Seong-Yun Jeong,3 Eun Kyung Choi,3,4 Tomomitsu Sewaki,5 Takeshi Arakawa,6,7 Shun’ichi Kuroda11Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan; 2College of Pharmacy, Duksung Women’s University, Seoul, South Korea; 3Institute for Innovative Cancer Research, ASAN Medical Center, Seoul, South Korea; 4Department of Radiation Oncology, University of Ulsan College of Medicine, Seoul, South Korea; 5GenoLac BL Corporation, Okinawa, Japan; 6COMB, Tropical Biosphere Research Center, 7Graduate School of Medicine, University of the Ryukyus, Okinawa, JapanAbstract: Dendritic cells (DCs) are key regulators of adaptive T-cell responses. By capturing exogenous antigens and presenting antigen-derived peptides via major histocompatibility complex molecules to naïve T cells, DCs induce antigen-specific immune responses in vivo. In order to induce effective host immune responses, active delivery of exogenous antigens to DCs is considered important for future vaccine development. We recently generated bionanocapsules (BNCs) consisting of hepatitis B virus surface antigens that mediate stringent in vivo cell targeting and efficient endosomal escape, and after the fusion with liposomes (LP) containing therapeutic materials, the BNC-LP complexes deliver them to human liver-derived tissues in vivo. BNCs were further modified to present the immunoglobulin G (IgG) Fc-interacting domain (Z domain) derived from Staphylococcus aureus protein A in tandem. When mixed with IgGs, modified BNCs (ZZ-BNCs) displayed the IgG Fv regions outwardly for efficient binding to antigens in an oriented-immobilization manner. Due to the affinity of the displayed IgGs, the IgG-ZZ-BNC complexes accumulated in specific cells and tissues in vitro and in vivo. After mixing ZZ-BNCs with antibodies against DCs, we used immunocytochemistry to examine which antibodies delivered ZZ-BNCs to mouse splenic DCs following intravenous injection of the ZZ-BNCs. ZZ-BNCs displaying anti-CD11c monoclonal antibodies (α-CD11c-ZZ-BNCs) were found to accumulate with approximately 62% of splenic DCs, and reside within some of them. After the fusion with liposomes containing antigens, the α-CD11c-ZZ-BNCs could elicit the respective antibodies more efficiently than other nontargeting control vaccines, suggesting that this DC-specific nanocarrier is promising for future vaccines.Keywords: drug-delivery system, gene-delivery system, liposomes, protein A, vaccine, ZZ domain