Molecular engineering and plant expression of an immunoglobulin heavy chain scaffold for delivery of a dengue vaccine candidate

• Published in: Plant biotechnology journal Vol. 15; no. 12; pp. 1590 - 1601
• Main Authors: Kim, Mi‐Young, Van Dolleweerd, Craig, Copland, Alastair, Paul, Matthew John, Hofmann, Sven, Webster, Gina R., Julik, Emily, Ceballos‐Olvera, Ivonne, Reyes‐del Valle, Jorge, Yang, Moon‐Sik, Jang, Yong‐Suk, Reljic, Rajko, Ma, Julian K.
• Format: Journal Article
• Language: English
• Published: England John Wiley & Sons, Inc 01.12.2017; John Wiley and Sons Inc
• Subjects: Animals; Antibody response; Antigen-presenting cells; Antigens; Autoantigens; CD4 antigen; CD8 antigen; Cellular manufacture; CHO Cells; Compartments; Complement component C1q; Conserved sequence; Cricetulus; Dengue; Dengue fever; Dengue Vaccines - administration & dosage; Dengue Vaccines - genetics; Dengue Vaccines - pharmacology; Dimers; Drug delivery systems; Engineering; Fc receptors; Female; Gene Expression Regulation, Plant; Glycoprotein E; Glycoproteins; High-performance liquid chromatography; Humans; IgG antibody; Immune response; Immune response (cell-mediated); Immune system; Immunogenicity; Immunoglobulin G; Immunoglobulin Heavy Chains - genetics; Immunoglobulin Heavy Chains - immunology; Immunoglobulins; infection; Liquid chromatography; Lymphocytes; Lymphocytes T; Macrophages; Macrophages - drug effects; Macrophages - metabolism; Male; Mice, Inbred BALB C; Molecular weight; Monomers; Neutralization Tests; Nicotiana - genetics; plant biotechnology; Plants, Genetically Modified - genetics; Polymers; Protein Domains; Protein Engineering - methods; Receptors; Recombinant Proteins - genetics; Recombinant Proteins - immunology; T-Lymphocytes - drug effects; T-Lymphocytes - immunology; Tobacco; vaccine; Vaccines; Vector-borne diseases; Viral diseases; Viruses; immunoglobulin G; infection; vaccine; plant biotechnology; dengue
• ISSN: 1467-7644; 1467-7652
• DOI: 10.1111/pbi.12741

Summary In order to enhance vaccine uptake by the immune cells in vivo, molecular engineering approach was employed to construct a polymeric immunoglobulin G scaffold (PIGS) that incorporates multiple copies of an antigen and targets the Fc gamma receptors on antigen‐presenting cells. These self‐adjuvanting immunogens were tested in the context of dengue infection, for which there is currently no globally licensed vaccine yet. Thus, the consensus domain III sequence (cEDIII) of dengue glycoprotein E was incorporated into PIGS and expressed in both tobacco plants and Chinese Ovary Hamster cells. Purified mouse and human cEDIII‐PIGS were fractionated by HPLC into low and high molecular weight forms, corresponding to monomers, dimers and polymers. cEDIII‐PIGS were shown to retain important Fc receptor functions associated with immunoglobulins, including binding to C1q component of the complement and the low affinity Fcγ receptor II, as well as to macrophage cells in vitro. These molecules were shown to be immunogenic in mice, with or without an adjuvant, inducing a high level IgG antibody response which showed a neutralizing potential against the dengue virus serotype 2. The cEDIII‐PIGS also induced a significant cellular immune response, IFN‐γ production and polyfunctional T cells in both the CD4+ and CD8+ compartments. This proof‐of‐principle study shows that the potent antibody Fc‐mediated cellular functions can be harnessed to improve vaccine design, underscoring the potential of this technology to induce and modulate a broad‐ranging immune response.