Expression of Norwalk virus capsid protein in transgenic tobacco and potato and its oral immunogenicity in mice

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 93; no. 11; pp. 5335 - 5340
• Main Authors: Mason, H.S. (Boyce Thompson Institute for Plant Research, Ithaca, NY.), Ball, J.M, Shi, J.J, Jiang, X, Estes, M.K, Arntzen, C.J
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 28.05.1996; National Acad Sciences
• Subjects: Administration, Oral; Animals; Antibodies; Antibodies, Viral - biosynthesis; Antibody Formation; ANTIGENE; ANTIGENOS; Base Sequence; CALICIVIRIDAE; Capsid - biosynthesis; Capsid - immunology; Capsid proteins; Cell Line; DNA; DNA Primers; EXPRESION GENICA; EXPRESSION DES GENES; FEUILLE; GENE; GENES; Hepatitis antigens; HOJAS; IMMUNISATION; IMMUNOGLOBULINE; INGESTION DE ALIMENTOS; INMUNIZACION; INMUNOGLOBULINA; Leaves; Mice; Mice, Inbred Strains; Molecular Sequence Data; Nicotiana - metabolism; NICOTIANA TABACUM; Norwalk virus; Norwalk virus - genetics; Norwalk virus - immunology; Norwalk virus - metabolism; PAPA; PLANTAS TRANSGENICAS; PLANTE TRANSGENIQUE; Plants, Genetically Modified; Plants, Toxic; Polymerase Chain Reaction; POMME DE TERRE; PRISE ALIMENTAIRE (HOMME); PROTEINAS; PROTEINE; RATON; Repetitive Sequences, Nucleic Acid; REPONSE IMMUNITAIRE; RESPUESTA INMUNOLOGICA; RNA; SOLANUM TUBEROSUM; Solanum tuberosum - metabolism; SOURIS; Spodoptera; Transfection; TRANSFERENCIA DE GENES; TRANSFERT DE GENE; TRANSFORMACION GENETICA; TRANSFORMATION GENETIQUE; TUBERCULE; TUBERCULO; Tubers; VACCIN SYNTHETIQUE; Vaccines, Synthetic - administration & dosage; VACUNA SINTETICA; Viral Vaccines - administration & dosage; VIRUS; Viruses
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.93.11.5335

Alternatives to cell culture systems for production of recombinant proteins could make very safe vaccines at a lower cost. We have used genetically engineered plants for expression of candidate vaccine antigens with the goal of using the edible plant organs for economical delivery of oral vaccines. Transgenic tobacco and potato plants were created that express the capsid protein of Norwalk virus, a calicivirus that causes epidemic acute gastroenteritis in humans. The capsid protein could be extracted from tobacco leaves in the form of 38-nm Norwalk virus-like particles. Recombinant Norwalk virus-like particle (rNV) was previously recovered when the same gene was expressed in recombinant baculovirus-infected insect cells. The capsid protein expressed in tobacco leaves and potato tubers cosedimented in sucrose gradients with insect cell-derived rNV and appeared identical to insect cell-derived rNV on immunoblots of SDS/polyacrylamide gels. The plant-expressed rNV was orally immunogenic in mice. Extracts of tobacco leaf expressing rNV were given to CD1 mice by gavage, and the treated mice developed both serum IgG and secretory IgA specific for rNV. Furthermore, when potato tubers expressing rNV were fed directly to mice, they developed serum IgG specific for rNV. These results indicate the potential usefulness of plants for production and delivery of edible vaccines. This is an appropriate technology for developing countries where vaccines are urgently needed.