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A Chlamydomonas-derived human papillomavirus 16 E7 vaccine induces specific tumor protection.

TitoloA Chlamydomonas-derived human papillomavirus 16 E7 vaccine induces specific tumor protection.
Tipo di pubblicazioneArticolo su Rivista peer-reviewed
Anno di Pubblicazione2013
AutoriDemurtas, Olivia Costantina, Massa Silvia, Ferrante Paola, Venuti Aldo, Franconi Rosella, and Giuliano Giovanni
RivistaPloS ONE
Volume8
Paginazionee61473
Data di pubblicazione2013
ISSN1932-6203
Parole chiave3' untranslated region, 3' Untranslated Regions, 5' untranslated region, 5' Untranslated Regions, algal extract, animal, animal cell, animal experiment, animal model, Animals, Antibodies, antigen specificity, article, binding affinity, biosynthesis, blood, C57BL mouse, cancer prevention, carcinogenesis, Cell Line, Chlamydomonas, Chlamydomonas extract, Chlamydomonas reinhardtii, chloroplast, chloroplast genome, Chloroplasts, clinical effectiveness, controlled study, E7GGG gene, E7GGG protein, Female, Gene expression, genetics, homologous recombination, human, Human papillomavirus, Human papillomavirus - 16, Human papillomavirus 16, Human Papillomavirus 16 E7 vaccine, Human papillomavirus type 16, Humans, hybrid protein, immunoglobulin G antibody, immunology, Inbred C57BL, lymphocyte proliferation, Mice, Mortality, mouse, Mus, nonhuman, nucleotide sequence, Papillomavirus E7 Proteins, Papillomavirus Infections, Papillomavirus Vaccines, protein E7, protein expression, protein purification, Recombinant Fusion Proteins, Subunit, subunit vaccine, survival, Survival Analysis, T lymphocyte, Tumor, tumor cell line, tumor immunity, unclassified drug, Uterine Cervical Neoplasms, uterine cervix cancer, Vaccines, Viral, virus antibody, virus oncogene, Wart virus vaccine
Abstract

BACKGROUND: The E7 protein of the Human Papillomavirus (HPV) type 16, being involved in malignant cellular transformation, represents a key antigen for developing therapeutic vaccines against HPV-related lesions and cancers. Recombinant production of this vaccine antigen in an active form and in compliance with good manufacturing practices (GMP) plays a crucial role for developing effective vaccines. E7-based therapeutic vaccines produced in plants have been shown to be active in tumor regression and protection in pre-clinical models. However, some drawbacks of in whole-plant vaccine production encouraged us to explore the production of the E7-based therapeutic vaccine in Chlamydomonas reinhardtii, an organism easy to grow and transform and fully amenable to GMP guidelines. METHODOLOGYPRINCIPAL FINDINGS: An expression cassette encoding E7GGG, a mutated, attenuated form of the E7 oncoprotein, alone or as a fusion with affinity tags (His6 or FLAG), under the control of the C. reinhardtii chloroplast psbD 5’ UTR and the psbA 3’ UTR, was introduced into the C. reinhardtii chloroplast genome by homologous recombination. The protein was mostly soluble and reached 0.12% of total soluble proteins. Affinity purification was optimized and performed for both tagged forms. Induction of specific anti-E7 IgGs and E7-specific T-cell proliferation were detected in C57BL/6 mice vaccinated with total Chlamydomonas extract and with affinity-purified protein. High levels of tumor protection were achieved after challenge with a tumor cell line expressing the E7 protein. CONCLUSIONS: The C. reinhardtii chloroplast is a suitable expression system for the production of the E7GGG protein, in a soluble, immunogenic form. The production in contained and sterile conditions highlights the potential of microalgae as alternative platforms for the production of vaccines for human uses.

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URLhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-84876587491&doi=10.1371%2fjournal.pone.0061473&partnerID=40&md5=d1407daadea89ae43fe2bc8a561a7812
DOI10.1371/journal.pone.0061473
Citation Key3987