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A plant protein signal sequence improved humoral immune response to HPV prophylactic and therapeutic DNA vaccines

TitleA plant protein signal sequence improved humoral immune response to HPV prophylactic and therapeutic DNA vaccines
Publication TypeArticolo su Rivista peer-reviewed
Year of Publication2017
AuthorsMassa, Silvia, Paolini F., Curzio G., Cordeiro M.N., Illiano E., Demurtas Olivia Costantina, Franconi Rosella, and Venuti A.
JournalHuman Vaccines and Immunotherapeutics
KeywordsAdaptive Immunity, amino acid composition, animal, animal experiment, animal model, Animals, Antibodies, antibody response, article, blood, C57BL mouse, capsid protein, Capsid Proteins, Cellular, cellular immunity, coat protein, controlled study, DNA, DNA vaccine, enzyme linked immunosorbent assay, enzyme linked immunospot assay, Female, gamma interferon, Gene expression, genetic transfection, genetics, human, human cell, Human papillomavirus type 16, Humoral, humoral immunity, Immunity, Immunofluorescence, immunoglobulin G, immunology, Inbred C57BL, kanamycin, L2 protein, membrane protein, Mice, mouse, nonhuman, oncogene protein E7, Oncogene Proteins, oncoprotein, Papillomavirus E7 Proteins, papillomavirus infection, Papillomavirus Infections, Papillomavirus Vaccines, plant protein, Plant Proteins, plasmid, polygalacturonase, Potato virus X, Potexvirus, prophylaxis, protein E7, Protein Sorting Signals, recombinant gene, recombinant protein, Recombinant Proteins, recombinant vaccine, signal peptide, Synthetic, treatment outcome, vaccination, Vaccines, Viral, virus antibody, virus antigen, Wart virus, Wart virus vaccine, Western blotting

Signal sequences (ss) play a critical role in the sorting of nascent secretory and membrane proteins. This function has been conserved from bacteria through eukaryotes, although ss appear diverse in length and amino acid composition. Sorting of proteins is also critical to instruct antigens for a proper immunological response. Thus, a plant ss was used to drive Human Papillomavirus (HPV) model antigens into the human secretory pathway: the HPV16 E7 oncoprotein, its chimera with the coat protein (CP) of the Potato Virus X (PVX), the first 200 amino acids of the HPV16 minor capsid protein L2 (known to harbour cross-reacting epitopes) and its chimera with E7 gene. These genes were used to transfect HEK-293 cells and to immunize C57BL/6 mice. The ss-provided genes were expressed, and proteins detected by immunofluorescence and immunoblotting. Mouse immunization with DNA constructs carrying the ss elicited a strong humoral response against both E7 and L2 and a weak cell-mediated immunity. To our knowledge this is the first demonstration that a signal sequence derived from a plant can modulate the sorting of a heterologous protein in mammalian cells. This activity in mammalian cells may be responsible for the observed increased humoral response to DNA-based vaccines that are generally weak inducers of IgG response. This might open new perspectives in the design of DNA vaccines, especially to counteract infections where a strong humoral response is needed. © 2017 Taylor & Francis.


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Citation KeyMassa2017271